WO2025237916A1 - Biomaterial composition comprising a polyhydroxybutyrate in particulate form - Google Patents

Abstract

The present invention relates to an injectable biomaterial and more particularly to a filler or an implant consisting of a composition comprising at least one biocompatible polymer and PHA particles. The present invention therefore relates to a composition comprising a biocompatible polymer and PHA particles. It also relates to PHA particles and to a method for obtaining same. It also relates to the method for preparing the injectable composition comprising a biocompatible polymer and PHA particles. It also relates to the methods for implanting the injectable composition comprising PHA particles.

Description

BIOMATERIAL COMPOSITION COMPRISING POLYHYDROXYBUTYRATE IN PARTICULAR FORM

[0001] The invention relates to the field of biomaterial formulations, more particularly in the medical and aesthetic fields. In these applications, the formulations must exhibit optimized properties in terms of rheology, taking into account, in particular, the characteristics of the medical device used for injection, the characteristics of the treated area, and the desired therapeutic or cosmetic effect, and also in terms of persistence to have the most lasting effect possible.

[0002] Human skin tissue is composed of fibrous proteins such as collagen and an extracellular matrix (ECM) containing fibronectin, laminin, and glycosaminoglycans (GAGs). When skin tissue is damaged, i.e., it has defects, for example due to aging, a general method is to restore function and form by injecting a filler material made of synthetic polymers or biological tissues into the affected area. This technique makes it possible to restore tissue, fill defects, improve wrinkles, or correct contours.

[0003] The raw materials of fillers for facial plastic surgery include autologous dermal implants, collagen or hyaluronic acid (HA).

[0004] To improve the performance of fillers, attempts have been made to manufacture fillers containing microspheres made of biopolymers with a slow decomposition rate. Among these, fillers containing polycaprolactone (PCL) are absorbed by the body. The decomposition rate is very low, which is why it is used as a semi-permanent filler. However, these products nevertheless have drawbacks, such as the generation of byproducts that can cause inflammatory reactions.

[0005] It is therefore necessary to develop filling compositions based on a new material which not only has high biocompatibility, immediate and lasting effects, but also has the advantage of generating a high-quality and properly organized neo-tissue.

[0006] Polyhydroxyalkanoates (PHAs) are diverse biopolyesters with a similar structure and different groups of side chains, synthesized by a variety of microorganisms. Due to their excellent biodegradability and Due to their biocompatibility, PHAs have been used for numerous applications, including medical implants, bioengineering, and regenerative medicine.

[0007] The preparation of PHA-based particles that can be incorporated into filling materials which are by design aqueous formulations presents many technical difficulties ranging from the difficulty of preparing particles with a size distribution compatible with the preparation of injectable formulations under conditions that allow the precision requirement necessary for wrinkle filling to be met.

[0008] Dermal fillers must possess rheological properties and cohesiveness suitable for aesthetic injections, while also being injectable through a fine needle, for patient comfort and practitioner precision. In particular, fillers based on particles or microparticles can present injectability problems manifesting as needle clogging or an injection force that is too great for the practitioner.

[0009] In patent application WO2009158724A2 filed in the name of TEPHA, microparticle compositions in the form of a suspension in a liquid injection vehicle, such as a surfactant solution (tween 80), are disclosed. With regard to injectability tests, microparticle compositions consisting of poly-4-hydroxybutyrate are more difficult to inject than compositions comprising the copolymer of poly(4-hydroxybutyrate-co-3-hydroxybutyrate). It is therefore necessary to reduce the particle content or to choose a less fine needle to make the product injectable.

[00010] Finally, in application WO2016074794A1 filed on behalf of MER.Z, injectable compositions comprising cross-linked hyaluronic acid, solid hydroxyapatite particles, and carboxymethylcellulose (CMC) are disclosed. This application specifies that to maintain good injectability of these compositions, it is necessary to add non-cross-linked CMC. The observed effects are superior when using CMC instead of the non-cross-linked hyaluronic acid commonly used to improve injectability.

[00011] It therefore remains necessary to prepare injectable compositions containing a biomaterial, such as cross-linked hyaluronic acid among others, and microparticles of polyhydroxyalkanoates (PHA) which are injectable and suitable for medical and aesthetic fields.

[00012] Surprisingly, the compositions according to the invention are easily injectable through fine needles (27G), while possessing remarkable viscoelastic properties and an inducing microparticle content adapted to obtain the desired filling effect. [00013] Administering the compositions according to the invention requires a moderate and entirely acceptable injection force and does not lead to needle clogging. Furthermore, while fillers suitable for medical and aesthetic applications must be easily injectable, it is also essential that these products, containing solid particles dispersed in an injectable biomaterial, remain stable over time. This means they must remain homogeneous, and sedimentation or particle aggregation must be avoided.

[00014] The compositions according to the invention make it possible to limit, and even avoid, the static sedimentation of the incorporated microparticles.

[00015] The present invention relates to an injectable biomaterial and more particularly to a filler or an implant made up of a composition comprising at least one biocompatible polymer and PHA particles.

[00016] The present invention therefore relates to a composition comprising a biocompatible polymer and PHA particles.

[00017] It also relates to PHA particles and their method of obtaining them.

[00018] It also relates to the process of preparing said injectable composition comprising a biocompatible polymer and PHA particles.

[00019] It also relates to methods of implanting said injectable composition comprising PHA particles.

[00020] In one embodiment, the composition according to the invention is in solvent-free form, in particular water-free, for example in lyophilized form.

[00021] In one embodiment, the composition is in the form of an injectable aqueous composition.

[00022] When the composition is in aqueous form the biocompatible polymer is in the form of a gel or hydrogel and the PHA particles are suspended in the gel.

[00023] In one embodiment, the composition is in the form of a bipartite composition of which one part comprises an aqueous polymeric phase and the other part comprises particles suspended in an organic phase miscible in the aqueous polymeric phase.

[00024] In one embodiment, the composition is in the form of a bipartite composition, one part of which comprises an aqueous polymeric phase and the other part comprises the particles in dry form.

[00025] In the context of this application, "hydrogel" means a polymeric gel consisting of a three-dimensional network made up of at least one polymer, capable of absorbing a large quantity of water or aqueous solution and which has particular rheological properties, especially in terms of viscosity and viscoelasticity. [00026] Said network can be constituted by grafting and/or chemical crosslinking by creating bonds between polymer chains, these bonds being covalent bonds.

[00027] Said network can also be obtained by transient physical interactions for example ionic, hydrophobic or hydrogen bonds.

[00028] When the composition is in aqueous form, the aqueous phase consists of water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS or a biological type buffer used in cell culture of the HEPES type ((hydroxyethyl)piperazinyl]ethanesulfonic acid)

[00029] Dissolving the biocompatible polymer or the composition according to the invention or in solvent-free form is carried out by adding water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS.

[00030] In one embodiment, the biocompatible polymer is dissolved by adding water or an aqueous saline solution, for example a phosphate buffer solution, for example PBS, further comprising at least one active ingredient, for example an antioxidant and/or a local anesthetic.

[00031] In the present application, the term "filler" or "filling product" refers to a product intended to be injected into the dermal or subcutaneous layer of the skin in order to visually improve the wrinkled area of the face, and to serve to maintain volume by itself without pharmacological action.

[00032] On the other hand, implants are biomaterials used to replace and repair damaged organs in the human body.

[00033] Therefore, although the filler and the implant have the same composition (biocompatible polymer, PHA particles), their application objectives and effects are different.

[00034] In one embodiment, said biocompatible polymer is a polysaccharide and/or a protein and/or a polypeptide alone or in mixture.

[00035] In the following text, the concentrations of biocompatible polymer are calculated with reference to the mass of the biocompatible polymer composition used, before the particles are suspended in the composition, i.e., with reference to the mass of the gel or hydrogel or the mass of the aqueous polymer phase. In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 200 mg/g.

[00036] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 75 mg/g. [00037] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 5 mg/g and 50 mg/g.

[00038] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 2 mg/g and 50 mg/g.

[00039] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 4 mg/g and 40 mg/g.

[00040] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 5 mg/g and 30 mg/g.

[00041] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 10 mg/g and 30 mg/g.

[00042] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is between 10 mg/g and 40 mg/g.

[00043] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 2 mg/g.

[00044] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 4 mg/g.

[00045] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 5 mg/g.

[00046] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 6 mg/g.

[00047] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 8 mg/g.

[00048] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 10 mg/g.

[00049] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 14 mg/g.

[00050] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 20 mg/g.

[00051] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 40 mg/g.

[00052] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 60 mg/g.

[00053] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 80 mg/g.

[00054] In one embodiment, the concentration of biocompatible polymer in the injectable aqueous composition is 100 mg/g. [00055] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of particles is between 2 mg/g and 200 mg/g.

[00056] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of particles is between 2 mg/g and 75 mg/g.

[00057] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of particles is between 5 mg/g and 50 mg/g.

[00058] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of particles is between 2 mg/g and 50 mg/g.

[00059] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is between

4 mg/g and 40 mg/g.

[00060] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is between

5 mg/g and 30 mg/g.

[00061] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is between 10 mg/g and 30 mg/g.

[00062] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is between 10 mg/g and 40 mg/g.

[00063] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 2 mg/g.

[00064] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 4 mg/g.

[00065] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 5 mg/g.

[00066] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 6 mg/g.

[00067] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 8 mg/g.

[00068] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 10 mg/g.

[00069] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 14 mg/g. [00070] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 20 mg/g.

[00071] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 40 mg/g.

[00072] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 60 mg/g.

[00073] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 80 mg/g.

[00074] In one embodiment, the concentration of biocompatible polymer in the gel or hydrogel before suspension of the particles is 100 mg/g.

[00075] In the following text, particle concentrations are given with reference to the total mass of the composition, namely the biomaterial after the particles have been suspended.

[00076] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 200 mg/g.

[00077] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 75 mg/g.

[00078] In one embodiment, the PHA particle content in the injectable aqueous composition is between 5 mg/g and 50 mg/g.

[00079] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 50 mg/g.

[00080] In one embodiment, the PHA particle content in the injectable aqueous composition is between 4 mg/g and 40 mg/g.

[00081] In one embodiment, the PHA particle content in the injectable aqueous composition is between 5 mg/g and 30 mg/g.

[00082] In one embodiment, the PHA particle content in the injectable aqueous composition is between 10 mg/g and 30 mg/g.

[00083] In one embodiment, the PHA particle content in the injectable aqueous composition is between 10 mg/g and 40 mg/g.

[00084] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 700 mg/g.

[00085] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 600 mg/g.

[00086] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 500 mg/g.

[00087] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 400 mg/g. [00088] In one embodiment, the PHA particle content in the injectable aqueous composition is between 2 mg/g and 300 mg/g.

[00089] In one embodiment, the PHA particle content in the injectable aqueous composition is between 10 mg/g and 300 mg/g.

[00090] In one embodiment, the PHA particle content in the injectable aqueous composition is between 50 mg/g and 300 mg/g.

[00091] In one embodiment, the PHA particle content in the injectable aqueous composition is between 50 mg/g and 400 mg/g.

[00092] In one embodiment, the PHA particle content in the injectable aqueous composition is between 50 mg/g and 500 mg/g.

[00093] In one embodiment, the PHA particle content in the injectable aqueous composition is between 50 mg/g and 600 mg/g.

[00094] In one embodiment, the PHA particle content in the injectable aqueous composition is 2 mg/g. [00095] In one embodiment, the PHA particle content in the injectable aqueous composition is 4 mg/g. [00096] In one embodiment, the PHA particle content in the injectable aqueous composition is 5 mg/g. [00097] In one embodiment, the PHA particle content in the injectable aqueous composition is 6 mg/g. [00098] In one embodiment, the PHA particle content in the injectable aqueous composition is 8 mg/g. [00099] In one embodiment, the PHA particle content in the injectable aqueous composition is 10 mg/g. [000100] In one embodiment, the PHA particle content in the injectable aqueous composition is 14 mg/g. [000101] In one embodiment, the PHA particle content in the injectable aqueous composition is 20 mg/g.

[000102] In one embodiment, the PHA particle content in the injectable aqueous composition is 40 mg/g.

[000103] In one embodiment, the PHA particle content in the injectable aqueous composition is 60 mg/g.

[000104] In one embodiment, the PHA particle content in the injectable aqueous composition is 80 mg/g.

[000105] In one embodiment, the PHA particle content in the injectable aqueous composition is 100 mg/g. [000106] In one embodiment, the PHA particle content in the injectable aqueous composition is 50 mg/g. [000107] In one embodiment, the PHA particle content in the injectable aqueous composition is 125 mg/g.

[000108] In one embodiment, the PHA particle content in the injectable aqueous composition is 150 mg/g.

[000109] In one embodiment, the PHA particle content in the injectable aqueous composition is 200 mg/g.

[000110] In one embodiment, the PHA particle content in the injectable aqueous composition is 250 mg/g.

[000111] In one embodiment, the PHA particle content in the injectable aqueous composition is 300 mg/g.

[000112] In one embodiment, the PHA particle content in the injectable aqueous composition is 350 mg/g.

[000113] In one embodiment, the PHA particle content in the injectable aqueous composition is 400 mg/g.

[000114] In one embodiment, the PHA particle content in the injectable aqueous composition is 450 mg/g.

[000115] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 200 mg/g.

[000116] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 75 mg/g.

[000117] In one embodiment, the PHA particle content in the biomaterial is between 5 mg/g and 50 mg/g.

[000118] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 50 mg/g.

[000119] In one embodiment, the PHA particle content in the biomaterial is between 4 mg/g and 40 mg/g.

[000120] In one embodiment, the PHA particle content in the biomaterial is between 5 mg/g and 30 mg/g.

[000121] In one embodiment, the PHA particle content in the biomaterial is between 10 mg/g and 30 mg/g.

[000122] In one embodiment, the PHA particle content in the biomaterial is between 10 mg/g and 40 mg/g.

[000123] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 700 mg/g.

[000124] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 600 mg/g.

[000125] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 500 mg/g. [000126] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 400 mg/g.

[000127] In one embodiment, the PHA particle content in the biomaterial is between 2 mg/g and 300 mg/g.

[000128] In one embodiment, the PHA particle content in the biomaterial is between 10 mg/g and 300 mg/g.

[000129] In one embodiment, the PHA particle content in the biomaterial is between 50 mg/g and 300 mg/g.

[000130] In one embodiment, the PHA particle content in the biomaterial is between 50 mg/g and 400 mg/g.

[000131] In one embodiment, the PHA particle content in the biomaterial is between 50 mg/g and 500 mg/g.

[000132] In one embodiment, the PHA particle content in the biomaterial is between 50 mg/g and 600 mg/g.

[000133] In one embodiment the PHA particle content in the biomaterial is 2 mg/g.

[000134] In one embodiment the PHA particle content in the biomaterial is 4 mg/g.

[000135] In one embodiment the PHA particle content in the biomaterial is 5 mg/g.

[000136] In one embodiment the PHA particle content in the biomaterial is 6 mg/g.

[000137] In one embodiment the PHA particle content in the biomaterial is 8 mg/g.

[000138] In one embodiment the PHA particle content in the biomaterial is 10 mg/g.

[000139] In one embodiment the PHA particle content in the biomaterial is 14 mg/g.

[000140] In one embodiment the PHA particle content in the biomaterial is 20 mg/g.

[000141] In one embodiment the PHA particle content in the biomaterial is 40 mg/g.

[000142] In one embodiment the PHA particle content in the biomaterial is 60 mg/g.

[000143] In one embodiment the PHA particle content in the biomaterial is 80 mg/g.

[000144] In one embodiment the PHA particle content in the biomaterial is 100 mg/g. [000145] In one embodiment, the PHA particle content in the biomaterial is 50 mg/g.

[000146] In one embodiment, the PHA particle content in the biomaterial is 125 mg/g.

[000147] In one embodiment, the PHA particle content in the biomaterial is 150 mg/g.

[000148] In one embodiment, the PHA particle content in the biomaterial is 200 mg/g.

[000149] In one embodiment, the PHA particle content in the biomaterial is 250 mg/g.

[000150] In one embodiment, the PHA particle content in the biomaterial is 300 mg/g.

[000151] In one embodiment, the PHA particle content in the biomaterial is 350 mg/g.

[000152] In one embodiment, the PHA particle content in the biomaterial is 400 mg/g.

[000153] In one embodiment, the PHA particle content in the biomaterial is 450 mg/g.

[000154] In one embodiment, the polysaccharide is selected from the group consisting of hyaluronic acid, keratin, heparin, cellulose, cellulose derivatives (in particular methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, ethylmethylcellulose, carboxymethylcellulose), alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparosan, and their biologically acceptable salts, alone or in mixture.

[000155] In one embodiment, said polysaccharide is hyaluronic acid.

[000156] In one embodiment, said polysaccharide is hyaluronic acid or one of its salts, alone or in a mixture. [000157] In one embodiment, said polysaccharide is hyaluronic acid in the form of a sodium or potassium salt. [000158] In one embodiment, said polysaccharide is hyaluronic acid in the form of a sodium salt.

[000159] In one embodiment, said polysaccharide is non-crosslinked hyaluronic acid or one of its salts, alone or in a mixture. [000160] In one embodiment, said polysaccharide is cross-linked hyaluronic acid or one of its salts, alone or in mixture. [000161] In one embodiment, said polysaccharide is cross-linked hyaluronic acid or one of its salts, mixed with non-cross-linked hyaluronic acid.

[000162] In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one crosslinking agent.

[000163] In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one bi- or polyfunctional crosslinking agent.

[000164] In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of at least one bifunctional crosslinking agent being butanedioldiglycidyl ether (BDDE) or 1,2,7,8-diepoxyoctane or divinylsulfone.

[000165] In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of BDDE.

[000166] In one embodiment, said polysaccharide is crosslinked hyaluronic acid or one of its salts, alone or in mixture, and in that said crosslinking is carried out by means of sodium or potassium trimetaphosphate.

[000167] When the polysaccharide, and more particularly hyaluronic acid, is crosslinked using a crosslinking agent, the degree of crosslinking (x) is calculated theoretically using the following formula: number of moles of crosslinking agent introduced into the reaction medium / number of moles of repeating units introduced into the reaction medium

[000168] With regard to hyaluronic acid, the repeating unit is a disaccharidic motif.

[000169] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.001 and 0.5.

[000170] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.001 and 0.4.

[000171] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.001 and 0.3.

[000172] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.001 and 0.2.

[000173] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.001 and 0.1. [000174] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.01 and 0.4.

[000175] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.01 and 0.5.

[000176] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.01 and 0.3.

[000177] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.1 and 0.2.

[000178] In one embodiment, the cross-linked hyaluronic acid has a cross-linking ratio x between 0.1 and 0.25

[000179] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.1 and 0.3.

[000180] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x between 0.1 and 0.35.

[000181] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.06.

[000182] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.07.

[000183] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.08.

[000184] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.09.

[000185] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.1.

[000186] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.11.

[000187] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.12.

[000188] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.13.

[000189] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.14.

[000190] In one embodiment, the crosslinked hyaluronic acid has a crosslinking ratio x of 0.15.

[000191] In one embodiment, said polysaccharide is co-crosslinked hyaluronic acid or one of its salts, alone or in mixture. [000192] In one embodiment, said polysaccharide is chemically modified hyaluronic acid by substitution, crosslinked or non-crosslinked, or one of its salts, alone or in mixture.

[000193] In one embodiment, said polysaccharide is doubly cross-linked hyaluronic acid as described in patent application W02000/046253 in the name of Fermentech medical limited.

[000194] In one embodiment, said polysaccharide is a mixture of hyaluronic acids, or one of their salts, crosslinked and non-crosslinked.

[000195] In one embodiment, said polysaccharide is a cross-linked mixture of hyaluronic acids, or one of their salts.

[000196] In one embodiment, said polysaccharide is a cross-linked mixture of hyaluronic acids, or one of their salts, such as that described in patent application W02009/071697 in the name of the applicant.

[000197] In one embodiment, said polysaccharide is a mixture of hyaluronic acids, obtained by mixing several hyaluronic acids, or one of their salts, of different molecular masses prior to their cross-linking, as described in patent application W02004092222 in the name of Cornéal industrie.

[000198] In one embodiment, said polysaccharide is hyaluronic acid or one of its salts, substituted by a group providing lipophilic or hydrating properties, such as, for example, substituted hyaluronic acids as described in patent application FR2983483 filed by the applicant. This application describes a process for the simultaneous substitution and crosslinking of a polysaccharide via its hydroxyl groups in an aqueous phase, characterized in that it comprises the following steps: (i) a polysaccharide is placed in an aqueous medium, (ii) it is brought into contact with at least one precursor of a substituent, (iii) it is brought into contact with a crosslinking agent, and (iv) the substituted and crosslinked polysaccharide is obtained and isolated. These process steps can be readily implemented within the scope of the present invention.

[000199] In one embodiment, said polysaccharide is hyaluronic acid or one of its salts, grafted with glycerol, for example as described in application WO2017162676 in the name of MERZ.

[000200] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 5 MDa (0.01 <Mw <5 MDa).

[000201] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 4 MDa (0.01 <Mw <4 MDa). [000202] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 3 MDa (0.01 <Mw <3 MDa).

[000203] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 2.5 MDa (0.01 <Mw <2.5 MDa).

[000204] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 2 MDa (0.01 <Mw <2 MDa).

[000205] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 1.5 MDa (0.01 <Mw < 1.5 MDa).

[000206] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.05 MDa to 2 MDa (0.05 <Mw <2 MDa).

[000207] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.1 MDa to 2 MDa (0.1 <Mw <2 MDa).

[000208]

[000209] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is within a range of 0.1 MDa to 3.5 MDa (0.1 <Mw <3.5 MDa).

[000210] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 2 MDa (0.5 <Mw <2 MDa).

[000211] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.75 MDa to 2 MDa (0.75 <Mw <2 MDa).

[000212] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 3 MDa (0.01 <Mw <3 MDa).

[000213] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 3 MDa (0.5 <Mw <3 MDa).

[000214] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is within a range of 1 MDa to 3 MDa (1 <Mw <3 MDa). [000215] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 4 MDa

(0.5 < Mw < 4 MDa).

[000216] In one embodiment, the weight-average molecular mass (Mw) of at least one hyaluronic acid is within a range of 0.75 MDa to

4 MDa (0.75 <Mw <4 MDa).

[000217] In one embodiment, the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 1 MDa to 4 MDa

(1 < Mw < 4 MDa)

[000218] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 0.5 MDa.

[000219] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 0.75 MDa.

[000220] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 1 MDa.

[000221] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 1.25 MDa.

[000222] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 1.5 MDa.

[000223] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 1.75 MDa.

[000224] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 2 MDa.

[000225] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 2.25 MDa.

[000226] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 2.5 MDa.

[000227] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 2.75 MDa.

[000228] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 3 MDa.

[000229] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 3.25 MDa.

[000230] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 3.5 MDa.

[000231] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 3.75 MDa. [000232] In one embodiment, the average molecular mass Mw of at least one hyaluronic acid is 4 MDa.

[000233] In one embodiment, the concentration of hyaluronic acid [HA] is between 2 mg/g and 50 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000234] In one embodiment, the concentration of hyaluronic acid [HA] is between 4 mg/g and 40 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000235] In one embodiment, the concentration of hyaluronic acid [HA] is between 5 mg/g and 30 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000236] In one embodiment, the concentration of hyaluronic acid [HA] is between 10 mg/g and 30 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000237] In one embodiment, the concentration of hyaluronic acid [HA] is 20 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000238] In one embodiment, the concentration of hyaluronic acid [HA] is 14 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000239] In one embodiment, the concentration of hyaluronic acid [HA] is 10 mg/g of total weight of said injectable aqueous composition before suspension of particles.

[000240] Furthermore, according to one embodiment of the invention, the composition comprises at least one protein and/or polypeptide alone or in mixture with polysaccharides.

[000241] According to one embodiment of the invention, the protein(s) and/or polypeptide(s) are selected from collagen, extracellular matrix proteins, structural proteins, blood-derived proteins, glycoproteins, fibronectins, laminins, lipoproteins, natural proteins, synthetic proteins, intracellular proteins, extracellular proteins, membrane proteins, and all combinations thereof.

[000242] According to one embodiment of the invention, the protein(s) and/or polypeptide(s) are selected from the group consisting of native human collagen, recombinant human collagen, recombinant humanized collagen, recombinant collagen-type proteins, native type 1 collagen from animals, for example from pigs, cattle, horses, native type 1 collagen from plants, by For example, orpine, and type O collagen derived from animals, for example jellyfish, alone or in mixtures

[000243] According to one embodiment of the invention, the collagen is selected from native collagen, collagen types I, II, III, V, XI, XXIV, collagen types VIII and X, collagen type IV, collagen type VI, collagen type VII, collagen types XIII, XVII, XXIII and XXV, collagen types XV and XVIII, collagen fibril la 1, collagen associated with fibrils types IX, XII, XIV, XVI, XIX, XX, XXI, XXII and XXVI and all their combinations.

[000244] Other forms of collagen may be used such as fibrillar atelopeptide collagen, telopeptide-containing collagen, freeze-dried collagen, mammalian collagen, recombinant collagen, reconstituted collagen, atelopeptide collagen obtained from a vertebrate species, recombinant collagen, collagen produced by genetically modified eukaryotic or prokaryotic cells or by genetically modified organisms, purified collagen and reconstituted purified collagen, collagen derived from genetically modified plants, proto-collagen and all their combinations.

[000245] Other forms of collagen may be used such as native human collagen, recombinant human collagen, recombinant humanized collagen, recombinant collagen-like proteins, alone or in mixture.

[000246] According to one embodiment of the invention, the collagen chosen is native human collagen, alone or in mixture.

[000247] According to one embodiment of the invention, the collagen chosen is recombinant human collagen, alone or in mixture.

[000248] According to one embodiment of the invention, the collagen chosen is recombinant humanized collagen, alone or in mixture.

[000249] According to one embodiment of the invention, the protein(s) and/or polypeptide(s) are recombinant collagen-type proteins, alone or in a mixture. [000250] The PHAs are selected from first-generation PHAs such as poly(3-hydroxybutyric acid) (PHB) and poly(4-hydroxybutyric acid) (P4HB), second-generation poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), third-generation poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx or PHBHx), fourth-generation poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB or P3HB4HB), or fifth-generation poly(3-hydroxybutyricacid-co-3-hydroxyvalericacid-co-3-hydroxyhexanoic acid) (PHBVHHx or PHBVHx), alone or in a mixture.

[000251] In one embodiment, PHA is P4HB. [000252] In one embodiment, PHA is poly(3-hydroxybutyrate-co-4-hydroxybutyrate).

[000253] In one embodiment, PHA is P4HB mixed with poly(3-hydroxybutyrate-co-4-hydroxybutyrate).

[000254] In one embodiment, the PHA particles have a size strictly less than 100 pm.

[000255] In one embodiment, the PHA particles have a size strictly less than 80 pm.

[000256] In one embodiment, the PHA particles have a size strictly less than 60 pm.

[000257] In one embodiment, the PHA particles have a size between 5 and 100 pm.

[000258] In one embodiment, the PHA particles have a size between 5 and 80 pm.

[000259] In one embodiment, the PHA particles have a size between 5 and 60 pm.

[000260] In one embodiment, the PHA particles have a size between 20 and 100 pm.

[000261] In one embodiment, the PHA particles have a size between 40 and 100 pm.

[000262] In one embodiment, the PHA particles have a size between 20 and 90 pm.

[000263] In one embodiment, the PHA particles have a size between 40 and 90 pm.

[000264] In one embodiment, the PHA particles have a size between 20 and 80 pm.

[000265] In one embodiment, the PHA particles have a size between 20 and 60 pm.

[000266] Control of particle size distribution during the particle formation process is fundamental because the PHA particles are incorporated into a biomaterial which is an injectable composition.

[000267] PHA particles are preferably prepared directly by a solvent evaporation technique, a double emulsion technique, or by microfluidization, using methods known from the literature. (Koosha, PhD thesis, 1989, Univ. Nottingham, UK, Diss. Abstr. Int. B 51: 1206 (1990); Bruhn & Müeller, Proceed Intern. Symp. Control. Rel. Bioact. Mater. 18:668-69 (1991); Conti, et al, J Microencapsulation, 9: 153-66 (1992); Ogawa, et al, Chem. Pharm. Bull., 36: 1095-103 (1988); Mathiowitz & Langer, "Polyanhydride microspheres as drug delivery systems," in Microcapsules Nanopart. Med. Pharm. (Donbrow, Ed.) ch. 5, pp. 99-123 (CRC, Boca Raton, Florida 1992).

[000268] In one embodiment, the processes for preparing PHA particles are chosen from solvent evaporation techniques of a dispersed phase of an emulsion, said emulsion being obtained conventionally by mixing, by ultrasonic emulsification or by membrane emulsification, but also from conventional spray-drying techniques or possibly electrostatic spraying and/or microfluidization.

[000269] In one embodiment, the particles are prepared by microfluidization using the droplet generation device described in patent application W02019/007965 in the name of ('UNIVERSITE LIBRE DE BRUXELLES', the contents of which are incorporated by reference.

[000270] In one embodiment, the particles are prepared by a membrane emulsification process.

[000271] In one embodiment, PHA is solubilized in an organic solvent that is immiscible and/or soluble in water, such as dichloromethane, for example at a concentration of between 1 and 10% of PHA in dichloromethane.

[000272] Water is added to the emulsion generation system to form droplets which are then collected, rinsed to remove solvents and then dried to obtain PHA particles.

[000273] In one embodiment, a surfactant or film-forming agent such as polyvinyl alcohol is added to the aqueous phase.

[000274] The dried and possibly calibrated particles by sieving are then incorporated according to various processes into the biocompatible polymer composition.

[000275] The PHA particles are suspended in water or in an aqueous saline solution, for example a phosphate buffer solution, for example PBS, and then this suspension is incorporated into a composition comprising a biocompatible polymer in solution or in the form of a gel or hydrogel in water or in an aqueous saline solution, for example a phosphate buffer solution, for example PBS, to obtain a composition according to the invention.

[000276] To preserve this composition, it can be lyophilized and then rehydrated with water or an aqueous saline solution, for example, a phosphate buffer solution, such as PBS, before injection. The invention also relates to the composition comprising the biocompatible polymer and PHA in lyophilized form. This composition must be rehydrated before use.

[000277] In one embodiment, the PHA particles are suspended in an organic liquid such as glycerol, polyethylene glycol, preferably having a molecular mass less than 1000 g.mol ¹ , or dimethyl sulfoxide

[000278] In one embodiment, the PHA particles are incorporated without having been suspended directly into an aqueous composition of biocompatible polymer in the form of a gel or hydrogel to obtain, by means of a mixing device, an injectable composition according to the invention.

[000279] In one embodiment the mixing device consists of two syringes connected at the time of incorporation and the mixing is carried out by making several back and forth movements until a homogeneous suspension is obtained.

[000280] In one embodiment, the mixing device is a conventional mixer and the incorporation of particles is carried out by mixing until a homogeneous composition is obtained.

[000281] Incorporation can be carried out just before use by the practitioner. Incorporation can also be carried out at the end of manufacturing, and the resulting composition will then be subjected to a lyophilization process to be rehydrated before injection.

[000282] In one embodiment, the composition according to the invention further comprises an active ingredient selected from the group consisting of local anesthetics, vitamin C derivatives, anti-inflammatories, antioxidants, antibiotics, bacteriostatics, antihemorrhagics or hemostatics and mixtures thereof.

[000283] In one embodiment, the composition according to the invention further comprises at least one local anesthetic.

[000284] In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 5%, relative to the total mass of said composition.

[000285] In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 4%, relative to the total mass of said composition.

[000286] In one embodiment, the composition according to the invention further comprises a local anesthetic at a local anesthetic concentration of between 0.1 and 2%, relative to the total mass of said composition.

[000287] In one embodiment, the composition according to the invention further comprises a local anesthetic to obtain a local anesthetic concentration of between 0.1 and 1%, relative to the total mass of said formulation.

[000288] In one embodiment, the composition according to the invention further comprises a local anesthetic to obtain a local anesthetic concentration of between 0.1 and 0.5%, relative to the total mass of said composition. [000289] In one embodiment, the composition according to the invention further comprises local anesthetic at a local anesthetic concentration of approximately 0.3%, relative to the total mass of said composition.

[000290] In one embodiment, the local anesthetic is chosen from the amino-ester group.

[000291] In one embodiment, the amino ester is chosen from the group comprising procaine, benzocaine, chloroprocaine and tetracaine in base or salt form, for example in hydrochloride form.

[000292] In one embodiment, the local anesthetic is chosen from the amino-amide group.

[000293] In one embodiment, the amino-amide is selected from the group comprising lidocaine, mepivacaine, prilocaine, articaine, aptocaine, bupivacaine, etidocaine and ropivacaine in base or salt form, for example in hydrochloride form.

[000294] In one embodiment, the local anesthetic is chosen from the amino-ether group.

[000295] In one embodiment, the amino-ether is chosen from the group comprising diamocaine and pramocaine in base or salt form, for example in hydrochloride or cyclamate form.

[000296] In one embodiment, the amino ether is selected from the group consisting of lidocaine, mepivacaine, and their isolated salts and isomers. [000297] In one embodiment, the amino ether is lidocaine.

[000298] In one embodiment, the amino-ether is lidocaine or one of its pharmaceutically acceptable salts.

[000299] In one embodiment, the amino ether is lidocaine hydrochloride. [000300] In one embodiment, the amino ether is mepivacaine.

[000301] In one embodiment, the amino-ether is mepivacaine or one of its pharmaceutically acceptable salts.

[000302] In one embodiment, the amino-ether is selected from the group consisting of racemic mepivacaine hydrochloride, racemic mepivacaine hydrochloride, (r)-mepivacaine hydrochloride, (s)-mepivacaine hydrochloride, (r)-mepivacaine and (s)-mepivacaine, or one of their pharmaceutically acceptable salts.

[000303] In one embodiment, the amino-ether is mepivacaine hydrochloride.

[000304] In one embodiment, the amino-ether is (r)-mepivacaine hydrochloride. [000305] In one embodiment, the amino-ether is (s)-mepivacaine hydrochloride.

[000306] In one embodiment, the amino-ether is racemic mepivacaine hydrochloride.

[000307] In one embodiment, the amino-ether is (r)-mepivacaine.

[000308] In one embodiment, the amino-ether is (s)-mepivacaine.

[000309] In one embodiment, the amino-ether is racemic mepivacaine.

[000310] In one embodiment, the composition according to the invention further comprises at least one amino-ether local anesthetic selected from the group consisting of lidocaine, mepivacaine, and mixtures thereof.

[000311] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine.

[000312] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 5%, relative to the total mass of said composition.

[000313] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 4%, relative to the total mass of said composition.

[000314] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 2%, relative to the total mass of said composition.

[000315] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 1%, relative to the total mass of said composition.

[000316] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of between 0.1 and 0.5%, relative to the total mass of said composition.

[000317] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely lidocaine, at a lidocaine concentration of approximately 0.3%, %, relative to the total mass of said composition.

[000318] In one embodiment, the composition according to the invention further comprises at least one local anesthetic, namely mepivacaine.

[000319] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 5%, relative to the total mass of said composition.

[000320] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 4%, relative to the total mass of said composition. [000321] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 2%, relative to the total mass of said composition.

[000322] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 1%, relative to the total mass of said composition.

[000323] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a concentration of mepivacaine between 0.1 and 0.5%, relative to the total mass of said composition.

[000324] In one embodiment, the composition according to the invention further comprises a local anesthetic being mepivacaine at a mepivacaine concentration of about 0.3%, relative to the total mass of said composition.

[000325] In one embodiment, the composition according to the invention further comprises at least one local anesthetic being diclonine in base or salt form, for example in hydrochloride form.

[000326] In one embodiment, the composition according to the invention further comprises at least one local anesthetic selected from the group consisting of chlorobutanol, guafecainol and polidocanol.

[000327] In one embodiment, the local anesthetic is chlorobutanol.

[000328] In one embodiment, the local anesthetic is guafecainol.

[000329] In one embodiment, the local anesthetic is polidocanol.

[000330] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory.

[000331] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of steroidal and non-steroidal anti-inflammatory drugs.

[000332] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory chosen from the group consisting of non-steroidal anti-inflammatory drugs.

[000333] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group comprising salicylate anti-inflammatories, propionic derivatives, indolic derivatives, pyrazole derivatives, oxicams and coxibs.

[000334] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group comprising diclofenac, nimesulfide, niflumic acid, mefenamic acid and nabumetone, alone or in mixture. [000335] In one embodiment, the composition according to the invention further comprises at least one salicylated anti-inflammatory selected from the group comprising diflunisal, benorilate and aspirin, alone or in mixture.

[000336] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of propionic derivatives including alminoprofen, ketoprofen, ibuprofen, naproxen, flurbiprofen and tiaprofenic acid, alone or in mixture.

[000337] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of indole derivatives including indomethacin, sulindac and etodolac, alone or in mixture.

[000338] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of pyrazole derivatives including in particular phenylbutazone.

[000339] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of oxicams comprising piroxicam, tenoxicam and meloxicam, alone or in mixture.

[000340] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group of coxibs comprising celecoxib, etoricoxib and rofecoxib, alone or in mixture.

[000341] In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.01 and 2000 mg/g. [000342] In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.1 and 1000 mg/g.

[000343] In one embodiment, the concentration of non-steroidal anti-inflammatory drugs in the composition according to the invention is between 0.5 and 500 mg/g.

[000344] In one embodiment, the composition according to the invention further comprises at least one steroidal anti-inflammatory.

[000345] In one embodiment, the composition according to the invention further comprises at least one steroidal anti-inflammatory selected from the group comprising dexamethasone, prednisolone, corticosterone, budesonide, sulfasalazine, mesalamine, cetirizine, diphenhydramine, antipyrine, methyl salicylate, loratadine, thymol, carvacrol, bisabolol, allantoin, eucalyptol, phenazone (antipyrine), propyphenazone, alone or in mixture.

[000346] In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.01 and 2000 mg/g. [000347] In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.1 and 1000 mg/g. [000348] In one embodiment, the concentration of steroidal anti-inflammatory drugs in the composition according to the invention is between 0.5 and 500 mg/g.

[000349] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of sucrose octasulfate and its salts.

[000350] In one embodiment, the composition according to the invention further comprises at least one anti-inflammatory selected from the group consisting of sucrose octasulfate and its sodium and potassium salts.

[000351] In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of alkali metal salts, alkaline earth metal salts, silver salts, ammonium salts, amino acid salts.

[000352] In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of alkali metal salts or alkaline earth metal salts.

[000353] In one embodiment, the composition according to the invention further comprises at least one water-soluble anti-inflammatory salt of sucrose octasulfate selected from the group consisting of sodium salt of sucrose octasulfate or potassium salt of sucrose octasulfate.

[000354] In one embodiment, the composition according to the invention further comprises at least one antimicrobial.

[000355] In one embodiment, the composition according to the invention further comprises at least one antimicrobial selected from the group comprising gentamicin, silver sulfadiazine, metronidazole, fucidin, bacitracin, eosin, povidone-iodine, copper gluconate, zinc gluconate, manganese gluconate or their salts, alone or in mixture.

[000356] In one embodiment, the concentration of antimicrobials in the composition according to the invention is between 0.1 and 200 mg/g.

[000357] In one embodiment, the concentration of antimicrobials in the composition according to the invention is between 0.5 and 100 mg/g.

[000358] In one embodiment, the composition according to the invention further comprises at least one glycoside or a glycoside derivative.

[000359] In one embodiment, the composition according to the invention further comprises at least one glycoside or a glycoside derivative selected from the group comprising D-glucopyranose, 1,4 glycoside, esculin, hesperidin, diosmin, arbutin, skimmine or aloin, alone or in mixture. Tl

[000360] In one embodiment, the concentration of glycosides in the composition according to the invention is between 0.1 and 200 mg/g.

[000361] In one embodiment, the concentration of glycosides in the composition according to the invention is between 0.5 and 100 mg/g.

[000362] , In one embodiment, the composition according to the invention further comprises at least antioxidant.

[000363] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of polyols.

[000364] In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol, propylene glycol, xylitol, glycerol, maltitol, lactitol and erythritol.

[000365] In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol, maltitol and glycerol, alone or in mixture.

[000366] In one embodiment, the composition according to the invention further comprises at least one polyol selected from the group consisting of mannitol, sorbitol and maltitol, alone or in mixture.

[000367] In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 0.1 mg/ml and 50 mg/ml, relative to the total mass of said composition.

[000368] In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000369] In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000370] In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000371] In one embodiment, the composition according to the invention further comprises at least one polyol at a polyol concentration of between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000372] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol.

[000373] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition. [000374] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000375] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000376] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being mannitol at a concentration between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000377] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol.

[000378] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration between 5 mg/ml and 40 mg/ml, relative to the total mass of said composition.

[000379] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 10 mg/ml and 40 mg/ml, relative to the total mass of said composition. [000380] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 20 mg/ml and 40 mg/ml, relative to the total mass of said composition. [000381] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being sorbitol at a concentration of between 30 mg/ml and 40 mg/ml, relative to the total mass of said composition. [000382] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being maltitol.

[000383] In one embodiment, biomaterial according to the invention further comprises at least one polyol, said polyol being glycerol.

[000384] In one embodiment, the composition according to the invention further comprises at least one polyol, said polyol being a mixture of mannitol and sorbitol.

[000385] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin C derivatives.

[000386] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin C derivatives including magnesium ascorbyl phosphate, sodium ascorbyl phosphate, ascorbyl-2-glucoside, and mixtures thereof.

[000387] In one embodiment, said at least one vitamin C derivative is magnesium ascorbyl phosphate. [000388] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of vitamin E derivatives and tocopherols.

[000389] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives.

[000390] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of carotenoids and retinoids and their derivatives including retinol, retinoic acid, retinal, retinol esters and carotene.

[000391] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of pseudo-tripeptides.

[000392] In one embodiment, the pseudo-tripeptide is glutathione.

[000393] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group comprising the various forms of coenzyme Q10, ubiquinone or ubiquinol.

[000394] In one embodiment, the composition according to the invention further comprises at least one vitamin.

[000395] In one embodiment, the composition according to the invention further comprises at least one vitamin selected from the group comprising retinol, thiamine, riboflavin, nicotinamide, dexpenthenol, piridoxine, ascorbic acid, ergocalciferol, tocopherol, biotin and folic acid alone or in mixture.

[000396] In one embodiment, the concentration of vitamins in the composition according to the invention is between 0.01 and 200 mg/g.

[000397] In one embodiment, the composition according to the invention further comprises at least one vasoconstrictor.

[000398] In one embodiment, the composition according to the invention further comprises at least one vasoconstrictor selected from the group comprising naphazoline, epinephrine, methoxamine, methylnorepinephrine, norepinephrine, oxymethazoline, phenylephrine, pseudoephedrine, synephrine, cirazolin and xylomethazoline.

[000399] In one embodiment, the concentration of vasoconstrictors in the composition according to the invention is between 0.01 and 3 mg/g.

[000400] In one embodiment, the composition according to the invention further comprises at least one vasodilator.

[000401] In one embodiment, the composition according to the invention further comprises at least one vasodilator selected from the group comprising adenosine, nicotinic acid, minoxidil and diazoxide, alone or in mixture. [000402] In one embodiment, the concentration of vasodilators in the composition according to the invention is between 0.01 and 10 mg/g.

[000403] In one embodiment, the composition according to the invention further comprises at least one anti-hemorrhagic or hemostatic agent.

[000404] In one embodiment, the composition according to the invention further comprises at least one antihemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, alone or in mixture.

[000405] In one embodiment, the concentration of antihemorrhagic or hemostatic agents in the composition according to the invention is between 0.01 and 5 mg/g. [000406] In one embodiment, the composition according to the invention further comprises at least one antioxidant and at least one local anesthetic.

[000407] In one embodiment, the composition according to the invention further comprises at least one antioxidant selected from the group of polyols and at least one local anesthetic selected from the group of amino-amides.

[000408] In one embodiment, the composition according to the invention further comprises at least one active ingredient selected from the group consisting of antibiotics.

[000409] In one embodiment, the composition according to the invention further comprises at least one antibiotic selected, for example, from the group of rifampicin, minocycline, gentamicin, fusidic acid, amoxicillin, azithromycin, benzathine, benzylpenicillin, cefaclor, cefadroxil, cephalexin, cefixime, cefotiam, cefpodoxime, ceftriaxone, cefuroxime, ciprofloxacin, clarithromycin, clindamycin, cloxacillin, cotrimoxazole, doxycycline, erythromycin, fosfomycin, josamycin, levofloxacin, metronidazole, minocycline, moxifloxacin, mupirocin, nitrofurantoin, norfloxacin, ofloxacin, ornidazole, phenoxymethylpenicillin (penicillin V), penicillin, pivmecillinam, pristinamycin, roxithromycin, spiramycin, sulfadiazine, tetracycline and trimethoprim, alone or in mixture.

[000410] In one embodiment, the composition according to the invention further comprises at least one mixture of antibiotic, for example the combination of rifampicin and minocycline.

[000411] In one embodiment, the composition according to the invention further comprises at least one mixture of antibiotic, for example the combination of gentamicin and minocycline.

[000412] In one embodiment, the composition according to the invention further comprises at least one anti-hemorrhagic or hemostatic agent.

[000413] In one embodiment, the composition according to the invention further comprises at least one antihemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, alone or in mixture. [000414] In one embodiment, the composition according to the invention is characterized in that it is injectable.

[000415] In one embodiment, the composition according to the invention is characterized in that it is sterile.

[000416] In one embodiment, the composition according to the invention is characterized in that it is injectable and sterile. [000417] The composition according to the invention has numerous applications. [000418] Among medical applications, examples include injections to replace deficient biological fluids, for example, in joints to replace synovial fluid, injections following surgery to prevent post-surgical adhesions, periurethral injections to treat incontinence, and injections following presbyopia surgery. Among aesthetic applications, examples include injections for filling wrinkles, fine lines, and skin defects, or for increasing volume, for example, of the lips, cheekbones, etc.

[000419] The applications targeted are more particularly those commonly used in the context of injectable viscoelastics and polysaccharides or collagen used or potentially usable in the following pathologies or treatments: aesthetic injections on the face: for filling wrinkles, skin defects or for volumizing (cheekbones, chin, lips); volumizing injections on the body: breast and buttock augmentation, G-spot augmentation, vaginoplasty, vaginal labia reconstruction, penis enlargement; in joint surgery and dental surgery for filling periodontal pockets, for example; treatment of osteoarthritis, injection into the joint to replace or supplement deficient synovial fluid; periurethral injection for the treatment of urinary incontinence due to sphincter insufficiency; post-surgical injection to prevent peritoneal adhesions in particular; injection following presbyopia surgery by laser scleral incisions; injection into the vitreous cavity; injection during cataract surgery; injection for the treatment of vaginal dryness; injection into tissue spaces; injection into the genitals. [000420] More particularly, in cosmetic surgery, depending on its viscoelastic and residual properties, the composition of the invention may be used: for filling fine, medium or deep wrinkles, and injected with fine diameter needles (27 Gauge for example); - as a volumizer with injection by needles of larger diameter, from 22 to 26 Gauge for example, and longer (30 to 40 mm for example); in this case, its cohesive character will ensure its retention at the injection site.

[000421] These examples of use are by no means limiting, and the composition of the invention can be used to: fill volumes; generate spaces within certain tissues, thus promoting their optimal functioning; replace deficient physiological fluids.

Examples

Particle preparation

Example A: Particle preparation

[000422] A solution of P4HB in dichloromethane with 5% P4HB is prepared and introduced into the device described in patent application W02019/007965 in the name of ('UNIVERSITE LIBRE DE BRUXELLES.

[000423] Water comprising 1% polyvinyl alcohol is introduced as a continuous phase into the above-mentioned device.

[000424] The flow rate is set at 50 pL/min and particles are obtained which are then washed with water to extract the solvent until it is eliminated and then dried under an airflow.

[000425] There is no coalescence and the beads are sufficiently solid to be handled after 30 minutes.

[000426] Drying is continued until completion and the particle size is measured, the diameter is 58 pm.

Example 1: Preparation of Phosphate/NaCl buffer solution

[000427] The buffer solution is prepared by introducing 0.225 g of NahUPCk^l-bO, 1.115 g of Na2HPC>4, and 42.5 g of NaCl into a 5 L volumetric flask which is filled to the mark with water for injection (WFI).

Example 2: Composition of non-crosslinked hyaluronic acid

[000428] The hyaluronic acid (HA) fibers used in the following examples 1 to 8 were purchased from HTL. They have an average molecular mass of 2.5 MDa and a moisture content of 13.7%.

[000429] 0.704 g of wet HA fibers (13.7%) are introduced into 20.30 g of buffer solution, to obtain a concentration of [non-crosslinked HA] of 30 mg.g ^-1 . The composition is hydrated overnight at room temperature.

[000430] The following day, the composition is mixed for 4 x 3 minutes with a rest period of 1.5 minutes between each mixing step. The composition is hydrated for 48 hours in a cold environment.

[000431] The composition is mixed again for 4 x 3 minutes with a rest period of 1.5 minutes between each mixing step. This is used as is in the following compositions. Example 3: Preparation of a composition including CMC and P4HB microbeads

[000432] A composition containing carboxymethylcellulose (CMC) is prepared by solubilizing 454 mg of CMC (AQUALON product type 7MF PH BET or 7LF PH BET degree of substitution 0.7 with a viscosity at 2% between 20 and 700 mPa.sec) in 20 mL of buffer solution prepared according to the protocol of example 1.

[000433] The resulting composition has a [CMC] concentration of 22.5 mg.g T

[000434] 162.2 mg of P4HB microbeads with a diameter of 58 pm obtained in Example A, are introduced into 4.09 mL of the previously obtained CMC composition to form a CMC composition comprising P4HB microbeads.

Example 4: Preparation of a composition including RH Collagen III and P4HB microbeads

[000435] A composition containing RH Collagen III is prepared by solubilizing 700 mg of RH Collagen III (DEMULCENT Product Purity>95%, collagen content 90-110%) in 10.3 mL of buffer solution prepared according to the protocol of Example 1. [000436] The composition obtained has a concentration of [RH Collagen III] of 68 mg.g ^-1 .

[000437] 173.2 mg of P4HB microbeads, with a diameter of 58 pm, obtained in Example A, are introduced into 4.33 mL of the buffer solution containing RH Collagen III to obtain an RH Collagen III composition comprising P4HB microbeads.

Example 5: Preparation of a composition comprising non-crosslinked hyaluronic acid and P4HB microbeads

[000438] A composition containing P4HB is prepared by suspending 79.5 mg of P4HB microbeads, 58 pm in diameter, obtained in Example A in 2 mL of buffer solution prepared according to the protocol of Example 1.

[000439] The composition obtained has a concentration in [P4HB] of 39.75 mg.g ^-1 .

[000440] 1 mL of this composition is mixed with 1.048 g of the non-crosslinked hyaluronic acid composition obtained in Example 2. To carry out the mixing, the two compositions are introduced into two 2.5 mL syringes which are connected together, and subsequently, back-and-forth movements are carried out until a homogeneous composition of non-crosslinked hyaluronic acid comprising P4HB microbeads is obtained. Example 6: Freeze-drying of compositions

[000441] Two grams of each of the compositions in Examples 3, 4 and 5 are taken and placed in the freeze dryer. Freeze drying is carried out for 12 h at a temperature of -55 °C, under a pressure of 100 mTorr.

[000442] After freeze-drying, the freeze-dried compositions are kept cold.

Example 7: Preparation of injectable compositions

[000443] The following non-limiting examples present distinct embodiments for preparing injectable compositions comprising different polysaccharides mixed with P4HB microbeads.

[000444] One embodiment consists of carrying out rehydration just before the injection of the mixtures obtained in example 6, using water for injection (WFI).

[000445] A second embodiment consists of carrying out a mixture just before the injection of an effective quantity of P4HB microbeads obtained in example A with an injectable composition comprising at least one polysaccharide.

[000446] A third embodiment consists of carrying out a mixture just before the injection of a suspension in the injectable water comprising P4HB microbeads obtained in example A with an injectable composition comprising at least one polysaccharide.

Example 7a

[000447] Composition 7a comprising RH Collagen III and P4HB microbeads, with a diameter of 58 pm, obtained in Example A is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 4 in 2 mL of water for injection.

[000448] This composition is divided into 4 portions. These are introduced into 4 1 mL syringes.

Example 7b

[000449] Composition 7b comprising non-crosslinked hyaluronic acid (HA) and P4HB microbeads, 58 pm in diameter, obtained in Example A is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 5 in 2 mL of water for injection.

[000450] This composition is divided into 3 portions. These are introduced into 3 1 mL syringes. Example 7c

[000451] Composition 7c comprising carboxymethylcellulose (CMC) and P4HB microbeads, with a diameter of 58 pm, obtained in Example A is prepared by hydrating 2 g of the lyophilized composition obtained in Example 6 from the composition obtained in Example 3 in 2 mL of water for injection.

[000452] This composition is divided into 2 portions. These are introduced into 2 1 mL syringes.

Example 7d

[000453] Composition 7d comprising STYLAGE XXL, marketed by VIVACY, prepared according to the process described in patent application W02009/071697, carboxymethylcellulose (CMC) and P4HB microbeads, 58 pm in diameter, obtained in Example A, is prepared by mixing the lyophilized composition obtained in Example 6 from the composition obtained in Example 3 and STYLAGE XXL in a mass ratio of 1:1.

[000454] This composition is introduced into a 1 mL syringe.

Example 7th

[000455] Compositions 7e comprising crosslinked hyaluronic acid and P4HB were made by mixing commercial products from the STYLAGE range, marketed by the company VIVACY, and P4HB in the form of solid microbeads having a diameter of 58 pm obtained in example A.

[000456] Each of the compositions 7e.1 to 7e.4 was prepared by mixing 1 mL of STYLAGE XXL product prepared according to the process described in patent application W02009/071697, contained in a syringe, and a defined mass of P4HB microbeads having a diameter of 58 pm obtained in Example A, contained in a second syringe, to obtain compositions having varying concentrations of [P4HB]. [000457] The two components are mixed by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension is obtained.

[000458] Each of the compositions 7e.5 to 7e.7 was prepared by mixing 0.8 mL of STYLAGE S product prepared according to the process described in patent application W02009/071697, contained in a syringe, and a defined mass of P4HB microbeads having a diameter of 58 pm obtained in Example A, contained in a second syringe, to obtain compositions having varying concentrations of [P4HB]. [000459] The two components are mixed by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension is obtained. [000460] The compositions and the various technical characteristics of compositions 7e.1 to 7e.7 are summarized in Table 1 below.

Table 1

Example 7f

[000461] A third embodiment consists of mixing just before injection a solution comprising P4HB microbeads obtained in example A with an injectable composition comprising at least one polysaccharide.

[000462] Compositions 7f comprising crosslinked hyaluronic acid and P4HB in the presence of a biocompatible solvent were made by mixing commercial products from the STYLAGE range, marketed by the company VIVACY, and an organic solvent containing P4HB microbeads having a diameter of 58 pm obtained in example A.

[000463] In a first step, a defined mass of P4HB microbeads is weighed and introduced into a defined volume of organic solvent and the suspension thus obtained is introduced into a syringe 1.

[000464] Each of the compositions 7f.l and 7f.2 were made by mixing 0.8 mL of STYLAGE S product prepared according to the process described in patent application W02009/071697, contained in a syringe 2, and the suspension obtained previously, contained in syringe 1, by connecting the two syringes end to end and making several back-and-forth movements until a homogeneous suspension was obtained.

[000465] The compositions and the various technical characteristics of compositions 7f.1 and 7f.2 are summarized in Table 2 below.

Table 2

Example 8: Injectability Tests

Example 8a

[000466] Composition 7a, comprising RH Collagen III and P4HB microbeads with a diameter of 58 µm, divided and introduced into four 1 mL syringes, was tested to evaluate its injectability. The results of these tests are presented in Table 3 below.

Table 3

[000467] Tests have shown that composition 7a is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced, and even in the presence of P4HB microbead agglomerates.

Example 8b

[000468] Composition 7b, comprising non-crosslinked HA and P4HB microbeads with a diameter of 58 µm, divided and introduced into three 1 mL syringes, was tested to evaluate its injectability. The results of these tests are presented in Table 4 below.

Table 4

[000469] Tests have shown that composition 7b is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced. Unlike previous cases, no agglomeration of P4HB microbeads is observed.

Example 8c

[000470] Composition 7c, comprising CMC and P4HB microbeads with a diameter of 58 µm, divided and introduced into two 1 mL syringes, was tested to evaluate its injectability. The results of these tests are presented in Table 5 below.

Table 5

[000471] Tests have shown that composition 7c is injectable regardless of the injection device used. It remains injectable even when the diameter of the needle used is significantly reduced, and even in the presence of P4HB microbead agglomerates.

Example 8d

[000472] Composition 7d, comprising STYLAGE XXL Lido prepared according to the process described in patent application W02009/071697, marketed by VIVACY, carboxymethylcellulose (CMC), and P4HB microbeads with a diameter of 58 µm, divided and introduced into two 1 mL syringes, was tested to evaluate its injectability. The results of these tests are presented in Table 6 below.

Table 6

[000473] The tests carried out show that composition 7d is injectable through a needle having a diameter of 27 G.

Example 8

[000474] The injectability of compositions 7e.1 to 7e.3, comprising the STYLAGE XXL product prepared according to the process described in patent application W02009/071697, marketed by the company VIVACY, and varying concentrations of P4HB microbeads, having a diameter of 58 pm, was evaluated and the results of these tests are presented in Table 7 below.

Table 7

[000475] The tests carried out show that the compositions 7e.1 to 7e.3 comprising cross-linked hyaluronic acid and varying concentrations of P4HB microbeads are injectable through needles having a diameter of 27 G, even when the concentration of microbeads increases sharply up to 99.7 mg.g ^-1 .

Example 8f

[000476] The injectability of compositions 7f.l and 7f.2, comprising the STYLAGE S product prepared according to the process described in patent application W02009/071697, marketed by the company VIVACY, of P4HB microbeads, having a diameter of 58 pm, and a biocompatible solvent was evaluated and the results of these tests are presented in Table 8, below.

Table 8

[000477] The tests carried out show that the compositions 7f.1 and 7f.2 comprising cross-linked hyaluronic acid, P4HB microbeads, at a concentration of approximately 100 mg.g ~ ¹ , and an organic solvent are injectable through needles having a diameter of 27 G.

Example 9: Preparation of a G1 cross-linked hyaluronic acid gel from 3 MDa sodium hyaluronate.

Step a: Hydration of sodium hyaluronate.

[000478] 114 g of sodium hyaluronate with an average molar mass by weight of approximately 3 MDa (injectable grade) and containing 12% residual moisture are dissolved at room temperature in an aqueous solution of 0.25 N sodium hydroxide under mechanical stirring.

Step b: Crosslinking.

[000479] 7.4 g of BDDE (99% purity) are diluted in a 0.25 N sodium hydroxide solution and then added to the sodium hyaluronate obtained in the previous step. The mixture is then homogenized in a paddle mixer. The reaction mixture, consisting of 100 g of dry sodium hyaluronate, 7.3 g of BDDE, and 782 g of 0.25 N sodium hydroxide, is placed in a water bath at 50 °C for 3 hours. The degree of crosslinking XI is approximately 0.13.

Step c: Neutralization, purification.

[000480] After crosslinking, the reaction medium is neutralized with a solution containing 1 N HCl and phosphate buffer supplemented with mannitol overnight at +4 °C under orbital stirring, then homogenized at room temperature by mechanical stirring.

[000481] The mannitol-supplemented phosphate buffer mentioned above was prepared by mixing the following compounds under mechanical stirring:

Table 9: Composition of phosphate buffer supplemented with mannitol.

[000482] The cross-linked hyaluronic acid gel obtained after mechanical agitation is loaded into a tubular dialysis membrane with a cutoff threshold of 12–14 kDa and dialyzed at +4 °C against mannitol-supplemented phosphate buffer as prepared above. The final sodium hyaluronate concentration is 27.5 mg/g, the pH is measured at 7.3, and the osmolarity at 292 mOsm/kg.

Example 10: Preparation of a G2 cross-linked hyaluronic acid gel from 1 MDa sodium hyaluronate.

[000483] A cross-linked hyaluronic acid gel was prepared according to a protocol similar to that previously described for the Gl gel, but from sodium hyaluronate of lower average weight molar mass.

[000484] In the crosslinking step, the reaction mixture composed of 100 g of dry sodium hyaluronate (of injectable grade and average molar mass by weight of approximately 1 MDa), 4.5 g of BDDE (99% purity), and 585 g of 0.25 N sodium hydroxide is placed in a water bath at 50 °C for 3 hours. The X2 crosslinking ratio is approximately 0.08.

[000485] At the end of the neutralization/purification step, the final concentration of sodium hyaluronate is 27.5 mg/g mg/g, the pH is measured at 7.1 and the osmolarity at 289 mOsm/kg.

Example 11: Preparation of a 20 mg/g G3 cross-linked hyaluronic acid gel. [000486] A 20 mg/g G3 cross-linked hyaluronic acid gel was prepared by diluting Gl gel with mannitol-supplemented phosphate buffer at room temperature. To do this, 200 g of 27.5 mg/g Gl gel were weighed into a container using a precision balance, and mannitol-supplemented phosphate buffer was added to Gl gel to make 275 g. The mixture was then manually stirred with a spatula using cycles of stirring and resting for a total of 15 minutes until a homogeneous mixture was obtained. The preparation is degassed by centrifugation at a Relative Centrifugal Acceleration (RCA) of 846 (2500 rpm) for 10 minutes on a Rotina 380 centrifuge (Andreas Hettich GmbH & co., Germany) equipped with a 6-place angular rotor with a rotation radius of 121 mm, then packaged in 1 mL BD Hypak™ SCF glass syringes before being sterilized by moist heat at 127 °C (F0 of 18.5 minutes). Example 12: Preparation of a G4 cross-linked hyaluronic acid gel at 20 mg/g. [000487] A G4 cross-linked hyaluronic acid gel was prepared at 20 mg/g by diluting the G2 gel with phosphate buffer supplemented with mannitol according to the protocol and in the same quantities as those described for the preparation of the G3 gel. It was then degassed by centrifugation, packaged, and then sterilized under the same conditions as the G3 gel (moist heat at 127 °C (F0 of 18.5 minutes).

Example 13: Preparation of non-crosslinked HA gels NR1 and NR2 at 20 mg/g Example 13a: NR1 preparation (LMW)

[000488] A low molecular weight hyaluronic acid gel (LMW NR1) at 20 mg/g is prepared using sodium hyaluronate fibers with a molecular weight of approximately 1.7 MDa and containing 10.3% residual moisture. 2.3 g of fibers are weighed into a plastic container using a precision balance. 97.1 g of mannitol-supplemented phosphate buffer are added to the fibers, and the mixture is then stirred using an IKA RW20 digital mechanical stirrer at 100 rpm for 18 hours at room temperature until the fibers are completely dissolved. The preparation is then degassed by centrifugation at a Relative Centrifugal Acceleration (RCA) of 846 (2500 rpm) for 10 minutes on a Rotina 380 centrifuge (Andreas Hettich GmbH & co., Germany,) and then packaged in 1 mL BD Hypak™ SCF glass syringes before being sterilized by moist heat at 127 °C (F0 of 18.5 minutes).

Example 13 b / Preparation NR2 (HMW)

[000489] A high molecular weight hyaluronic acid gel (HMW2) at 20 mg/g is prepared using sodium hyaluronate fibers with a molecular weight of approximately 3 MDa and a residual moisture content of 12.4%. 3.1 g of fibers are weighed into a plastic container using a precision balance. 118.7 g of phosphate buffer supplemented with mannitol are added to the fibers, and the mixture is then stirred using an IKA RW20 digital mechanical stirrer at 100 rpm for 18 hours at room temperature until the fibers are completely dissolved. The preparation is then degassed by centrifugation at a Relative Centrifugal Acceleration (RCA) of 846 (2500 rpm) for 10 minutes on a Rotina 380 centrifuge (Andreas Hettich GmbH & co., Germany) and then packaged in 1mL BD Hypak™ SCF glass syringes before being sterilized by moist heat at 127 °C (F0 of 18.5 minutes). Example 14: Preparation of PI microspheres of poly-4-hydroxybutyrate (P4HB)

[000490] Poly-4-hydroxybutyrate (P4HB) microspheres were prepared according to a membrane emulsification process.

[000491] 11.25 g of P4HB obtained by bacterial fermentation with a molar mass by weight ( _Mw ) of approximately 180 kDa were dissolved in 225 mL of dichloromethane to obtain a 5% (w/v) solution. In parallel, 4.275 L of a 2% (w/w) solution of polyvinyl alcohol (PVA 23-88) in deionized water was prepared.

[000492] The emulsion was produced at a flow rate of 475 mL/min for the aqueous phase and 25 mL/min for the organic phase using a 5 x 100 µm membrane and a 9.5 mm insert. The sample was produced and collected on an AXF-1 instrument using a digital collection pump system, ensuring a 5% (w/v) concentration of the organic phase during emulsification through a closed-loop delivery system.

[000493] During preparation, the emulsion was gradually collected in a beaker containing 1 L of 2% (w/w) PVA solution and kept under constant stirring at 200 rpm until the membrane extrusion process was complete. 4.5 L of emulsion were recovered before the addition of 4.5 L of deionized water for dilution. After the dilution step, the mixing speed was increased to 600 rpm and the height of the stirring paddle was adjusted to one-third of the height of the volume occupied by the mixture. Stirring was maintained for 96 hours before being stopped, sedimentation was performed, approximately 2 L of clear supernatant was removed, the mixture was filtered through an 8 µm membrane to remove particles smaller than the desired size range, and several rinses were performed with deionized water. The recovered particles were then lyophilized for approximately 24 hours. The lyophilized particles were then sieved through a 200 mm diameter Retsch® woven wire mesh sieve with a 75 µm mesh opening. The sieved microspheres were deposited onto a glass slide and dispersed in three drops of water before being observed under a light microscope. The appearance, size, and size distribution of the microspheres were determined using a ZEISS Axio Scope Al optical microscope with an Epiplan-Neofluar 20x/0.50 DIC M27 EC objective. Size was determined from a sample of 50 microspheres by taking two diameter measurements per microsphere.

[000494] The results are presented in Table 10 below.

Table 10: Size and size distribution of PI microspheres.

[000495] Example 15: Preparation of compositions A1-A4

[000496] Poly-4-hydroxybutyrate (P4HB) microspheres (PI) were used to prepare 4 g of composition by mixing them with G4 cross-linked hyaluronic acid gel, in order to produce compositions with microsphere concentrations of 5% (w/w) for composition A1, 10% (w/w) for composition A2, 15% (w/w) for composition A3, and 30% (w/w) for composition A4. To do this, a mass of PO microspheres was weighed directly into a 15 mL Falcon conical centrifuge tube. A mass of G4 cross-linked hyaluronic acid gel was then weighed and added to the microspheres to obtain a total composition mass of 4 g. The mixture was manually blended with a spatula at room temperature using agitation/rest cycles for a total duration of 10 minutes until a homogeneous composition was obtained.

[000497] Example 16: Preparation of compositions B1-B3

[000498] Compositions B1-B3 were produced according to a protocol identical to compositions A1-A4 by mixing poly-4-hydroxybutyrate (P4HB) microspheres - PI with the gel containing crosslinked hyaluronic acid - G3 in order to produce compositions having a microsphere concentration of 5% (w/w) for composition B1, 10% (w/w) for composition B2 and 15% (w/w) for composition B3.

[000499] The characteristics of the compositions are presented in Table 11 below.

Table 11: Composition of compositions G4, G3, Al to A4 and Bl to B3 Example 17: Preparation of compositions C1-C3 and D1-D4

[000500] Compositions C1-C3 and D1-D4 were produced according to a protocol identical to compositions A1-A4 by mixing poly-4-hydroxybutyrate (P4HB)-PI microspheres with NR1 and NR2 non-crosslinked hyaluronic acid-containing gels to produce compositions with a microsphere concentration of 5% (w/w) for compositions Cl and D1, 10% (w/w) for compositions C2 and D2 and 15% (w/w) for compositions C3 and D3.

[000501] The characteristics of the compositions are presented in Table 12 below.

Table 12 Composition of Cl to C3 and Dl to D3 compositions

Example 18: Determination of the resistance to sedimentation of the compositions according to the invention Example 18 a: Centrifugation.

[000502] After obtaining, all compositions are directly centrifuged in order to evaluate their resistance to sedimentation.

[000503] The compositions are centrifuged successively according to a Relative Centrifugal Acceleration (RCA) of 846 (2500 rpm), 3382 (5000 rpm) and 7610 (7500 rpm) for 10 minutes respectively on a Rotina 380 centrifuge (Andreas Hettich GmbH & co., Germany) equipped with a 6-place angular rotor with a rotation radius of 121 mm.

[000504] Between each centrifugation condition, the compositions are manually mixed with a spatula according to agitation/rest cycles at room temperature for a total duration of 10 minutes until a homogeneous composition is obtained.

Example 18b: Static storage.

[000505] After this accelerated sedimentation study, the compositions are remixed before being packaged in 1mL BD Hypak™ SCF glass syringes and stored at 4 °C.

[000506] The compositions packaged in syringes are then observed after different storage times at 4 °C in order to assess their homogeneity and the presence of any sedimentation. [000507] In both analyses, the presence of sedimentation is assessed by visual inspection. Sedimentation occurs when a deposit of particles is visible to the naked eye at the bottom of the container.

[000508] The results obtained are presented in Table 13 below.

Table 13: Resistance to sedimentation of the different compositions

[000509] Tests carried out under static storage conditions have demonstrated that P4HB microspheres dispersed in a cross-linked sodium hyaluronate composition do not sediment after at least 35 days of storage at 4 °C.

[000510] Sedimentation is observed after 10 min of centrifugation according to an ACR of 7610 for compositions A1, A2, A3 and A4 comprising a crosslinked sodium hyaluronate gel with a crosslinking ratio of 0.08 prepared from a 1 MDa sodium hyaluronate. Under equivalent centrifugation conditions, no sedimentation is observed for compositions B1, B2 and B3 comprising a crosslinked sodium hyaluronate gel with a crosslinking ratio of 0.13 prepared from a 3 MDa sodium hyaluronate gel.

Example 19: Determination of the resistance to sedimentation of comparative compositions.

[000511] The resistance to sedimentation of comparative compositions comprising non-crosslinked hyaluronic acid and increasing contents of P4HB microspheres was evaluated from the centrifugation tests and observations of syringes stored statically at 4 °C described previously.

[000512] The results obtained are presented in Table 14 below.

Table 14: Resistance to sedimentation of the different compositions

[000513] Under static storage conditions at 4 °C, sedimentation was observed for compositions Cl, C2 and C3 comprising a non-crosslinked 1 MDa sodium hyaluronate gel and PH4B microspheres. Under equivalent storage conditions, no sedimentation was observed in compositions Al, A2, A3 and A4 comprising a crosslinked hyaluronate gel (X2 = 0.08) obtained from a 1 MDa sodium hyaluronate; similarly, no sedimentation was observed in compositions Bl, B2 and B3 comprising a crosslinked hyaluronate gel (XI = 0.13) obtained from a 3 MDa sodium hyaluronate.

[000514] Also, after 10 minutes of centrifugation according to an ACR of 3382, sedimentation is observed for compositions Cl, C2 and C3 which is not the case for compositions DI, D2 and D3.

[000515] Furthermore, after 10 minutes of centrifugation according to an ACR of 7610, sedimentation was observed for compositions Cl, C2 and C3 and DI, D2 and D3 comprising a non-crosslinked 3 MDa sodium hyaluronate gel and PH4B microspheres. Under equivalent centrifugation conditions, no sedimentation was observed in compositions Bl, B2 and B3 comprising a crosslinked hyaluronate gel (XI = 0.13) obtained from a 3 MDa sodium hyaluronate.

Example 20: Determination of the rheological properties of the compositions according to the invention.

[000516] The rheological properties of the compositions were measured at a temperature of 25 °C on a Kinexus Prime Pro+ rheometer (Netzsch Group, Germany) with a smooth planar geometry of 20 mm diameter and a gap of 1 mm, by performing a strain sweep from ^10⁻³ % to ^10³ % at a frequency of 1 Hz. The rheological properties of the compositions, as described in application WO2022/038156, are defined as follows:

Elastic modulus (G') in the Linear Viscoelastic Region (LVER) in Pa; Viscous modulus (G") in the LVER in Pa;

Damping factor (tan ô) in the LVER;

LVER limit strain (yp) in % being defined as the strain for a 10% loss of the elastic modulus compared to the value measured in the LVER;

LVER boundary stress (TP) in Pa;

Elastic modulus at the crossing point (G'c) in Pa;

Threshold strain (yc) in %;

Threshold stress (TC) in Pa; - Plastic range in deformation (yc - yp) in %;

Plastic domain in Stress (TC - TP) in Pa.

[000517] The rheological properties of the compositions described above are summarized in Table 15 below:

Table 15: Viscoelastic properties measured on the compositions

[000518] For compositions comprising the crosslinked hyaluronic acid composition with a crosslinking ratio X2 = 0.08, an increase in G' is observed, as well as a strong increase in the extent of the plastic domain under stress and the extent of the plastic domain under strain.

[000519] Some of these results are also obtained for compositions including the crosslinked hyaluronic acid composition with a crosslinking ratio XI = 0.13, for which an increase in G' is noted, as well as an increase in the extent of the plastic domain under stress and a limited increase in the extent of the plastic domain under strain.

[000520] Compositions comprising a cross-linked hyaluronic acid gel and P4HB microspheres exhibit an increased elastic modulus G' compared to the equivalent cross-linked hyaluronic acid gel, in the absence of microspheres. Also, compositions comprising a cross-linked hyaluronic acid gel and P4HB microspheres exhibit larger plastic domains (under stress and strain) compared to compositions comprising equivalent hyaluronate gels, without microspheres.

Example 21: Determination of the injectability (injection force) of the compositions according to the invention.

[000521] The injectability of each formulation is determined on a Mecmesin Multitest li force test bench (Mecmesin/PPT Group, France) by measuring the average force required to extrude the sample at a syringe piston travel speed of 13 mm/min using 1 ml BD Hypak™ SCF glass syringes fitted with 27G x ^1/2 " (13 mm) ETW TSK needles. The analysis is performed by extruding a portion of the syringe contents corresponding to a syringe piston travel of 24 mm. The injectability of the formulation is then defined as the average extrusion force in Newtons for a piston travel between 5 mm and 20 mm. [000522] The results obtained are presented in Table 16 below.

Table 16: Injectability properties measured on the compositions [000523] The average injection force as a function of the concentration of P4HB microspheres in the cross-linked hyaluronic acid compositions is acceptable as it ranges between 17 and 35 N through a 27G needle. All the compositions tested are therefore injectable.

[000524] The compositions of the invention comprise a cross-linked hyaluronic acid gel and P4HB microspheres. These exhibit high elastic moduli, a wide plastic range, and are easily injectable through fine 27G needles. Furthermore, these compositions remain homogeneous over time when stored under static conditions in syringes.

Claims

Demands [Claim 1]. Injectable biomaterial consisting of a composition comprising at least one biocompatible polymer and PHA particles. [Claim 2]. Biomaterial according to claim 1, characterized in that the composition is in solvent-free form, in particular water-free, for example in lyophilized form. [Claim 3]. Biomaterial according to claim 1, characterized in that the composition is in aqueous form, the biocompatible polymer is in gel or hydrogel form, and the PHA particles are suspended in the gel. [Claim 4]. Biomaterial according to any one of the preceding claims, characterized in that said biocompatible polymer is a polysaccharide and/or a protein and/or a polypeptide alone or in mixture [Claim 5]. Biomaterial according to any one of the preceding claims, characterized in that the concentration of biocompatible polymer in the injectable aqueous composition before suspension of particles in the composition is between 2 mg/g and 75 mg/g. [Claim 6]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 200 mg/g. [Claim 7]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 700 mg/g. [Claim 8]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 600 mg/g. [Claim 9]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 500 mg/g. [Claim 10]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 400 mg/g. [Claim 11]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 2 mg/g and 300 mg/g. [Claim 12]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 10 mg/g and 300 mg/g. [Claim 13]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 50 mg/g and 300 mg/g. [Claim 14]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 50 mg/g and 400 mg/g. [Claim 15]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 50 mg/g and 500 mg/g. [Claim 16]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is between 50 mg/g and 600 mg/g. [Claim 17]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 50 mg/g. [Claim 18]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 125 mg/g. [Claim 19]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 150 mg/g. [Claim 20]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 200 mg/g. [Claim 21]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 250 mg/g. [Claim 22]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 300 mg/g. [Claim 23]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 350 mg/g. [Claim 24]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 400 mg/g. [Claim 25]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particle content in the injectable aqueous composition is 450 mg/g. [Claim 26]. Biomaterial according to any one of the preceding claims, characterized in that the polysaccharide is selected from the group consisting of hyaluronic acid, keratane, heparin, cellulose, cellulose derivatives (in particular hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylmethylcellulose, carboxymethylcellulose), alginic acid, xanthan gum, carrageenan, chitosan, chondroitin, heparosan, and their biologically acceptable salts, alone or in mixture. [Claim 27]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide is hyaluronic acid or one of its salts, alone or in mixture. [Claim 28]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide is cross-linked hyaluronic acid or one of its salts, alone or in mixture. [Claim 29]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide is cross-linked hyaluronic acid or one of its salts, in mixture with non-cross-linked hyaluronic acid. [Claim 30]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass Mw of at least one hyaluronic acid is within a range of 0.01 MDa to 5 MDa (0.01 <Mw <5 MDa). [Claim 31]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 4 MDa (0.01 <Mw <4 MDa). [Claim 32]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 3 MDa (0.01 <Mw <3 MDa). [Claim 33]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 2.5 MDa (0.01 <Mw <2.5 MDa). [Claim 34]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has an average molecular weight in weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 2 MDa (0.01 <Mw <2 MDa). [Claim 35]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 1.5 MDa (0.01 <Mw < 1.5 MDa). [Claim 36]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.05 MDa to 2 MDa (0.05 <Mw <2 MDa). [Claim 37]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.1 MDa to 2 MDa (0.1 <Mw <2 MDa). [Claim 38]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 2 MDa (0.5 <Mw <2 MDa). [Claim 39]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.75 MDa to 2 MDa (0.75 <Mw <2 MDa). [Claim 40]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.75 MDa to 2 MDa (0.75 <Mw <2 MDa). [Claim 41]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 5 MDa (0.5 <Mw <4 MDa). [Claim 42]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.75 MDa to 4 MDa (0.75 <Mw <4 MDa). [Claim 43]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 1 MDa to 4 MDa (1 <Mw <4 MDa). [Claim 44]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.5 MDa to 3 MDa (0.5 <Mw <3 MDa). [Claim 45]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the average molecular mass by weight (Mw) of at least one hyaluronic acid is within a range of 0.01 MDa to 3 MDa (0.01 <Mw <3 MDa). [Claim 46]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 0.5 MDa. [Claim 47]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 0.75 MDa. [Claim 48]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 1.25 MDa. [Claim 49]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 1.5 MDa. [Claim 50]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 1.75 MDa. [Claim 51]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 2 MDa. [Claim 52]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 2.25 MDa. [Claim 53]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 2.5 MDa. [Claim 54]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 2.75 MDa. [Claim 55]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 3.25 MDa. [Claim 56]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 3.5 MDa. [Claim 57]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 3.75 MDa. [Claim 58]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a weight average molecular mass (Mw) of at least one hyaluronic acid of 4 MDa. [Claim 59]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.001 and 0.5. [Claim 60]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.001 and 0.4. [Claim 61]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.001 and 0.3. [Claim 62]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.001 and 0.2. [Claim 63]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.001 and 0.1. [Claim 64]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.01 and 0.5. [Claim 65]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.01 and 0.4. [Claim 66]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.01 and 0.3. [Claim 67]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.01 and 0.2. [Claim 68]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.01 and 0.15. [Claim 69]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.1 and 0.2. [Claim 70]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a degree of crosslinking (X) of at least one crosslinked hyaluronic acid between 0.1 and 0.25. [Claim 71]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.1 and 0.3. [Claim 72]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide the degree of crosslinking (X) of at least one crosslinked hyaluronic acid is between 0.1 and 0.35. [Claim 73]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.08. [Claim 74]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.09. [Claim 75]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.1. [Claim 76]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.11. [Claim 77]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.13. [Claim 78]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.14. [Claim 79]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has a crosslinking ratio (X) of at least one crosslinked hyaluronic acid of 0.15. [Claim 80]. Biomaterial according to any one of the preceding claims, characterized in that said polysaccharide has an acid concentration hyaluronic [HA] is between 2 mg/g and 50 mg/g of total weight of said injectable aqueous composition. [Claim 81]. Biomaterial according to any one of the preceding claims, characterized in that the composition comprises at least one protein and/or polypeptide alone or in mixture with polysaccharides. [Claim 82]. Biomaterial according to claim 81, characterized in that the protein(s) and/or polypeptide(s) are selected from collagen, extracellular matrix proteins, structural proteins, blood-derived proteins, glycoproteins, fibronectins, laminins, lipoproteins, natural proteins, synthetic proteins, intracellular proteins, extracellular proteins, membrane proteins, and all combinations thereof. [Claim 83]. Biomaterial according to any one of the preceding claims, characterized in that the protein(s) and/or polypeptide(s) are selected from the group consisting of native human collagen, recombinant human collagen, recombinant humanized collagen, recombinant collagen-like proteins, native type 1 collagen from animals, e.g. pig, bovine, equine, native type 1 collagen from plants, e.g. oprin, and type 0 collagen from animals, e.g. jellyfish, alone or in mixture. [Claim 84]. Biomaterial according to any one of the preceding claims, characterized in that the protein(s) and/or polypeptide(s) is native human collagen, alone or in mixture. [Claim 85]. Biomaterial according to any one of the preceding claims, characterized in that the protein(s) and/or polypeptide(s) is recombinant human collagen, alone or in mixture. [Claim 86]. Biomaterial according to any one of the preceding claims, characterized in that the protein(s) and/or polypeptide(s) is recombinant humanized collagen, alone or in mixture. [Claim 87]. Biomaterial according to any one of the preceding claims, characterized in that the protein(s) and/or polypeptide(s) are recombinant collagen-type proteins, alone or in mixture. [Claim 88]. Biomaterial according to any one of the preceding claims, characterized in that the PHAs are selected from first-generation PHAs such as poly(3-hydroxybutyric acid) (PHB) and poly(4-hydroxybutyric acid) (P4HB), second-generation PHAs such as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), third-generation PHAs such as poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx or PHBHx), fourth-generation PHAs such as poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB or P3HB4HB), or fifth-generation PHAs such as poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB), or fifth-generation PHAs such as poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB). generation such as poly(3-hydroxybutyricacid-co-3-hydroxyvalericacid-co-3-hydroxyhexanoic acid) (PHBVHHx or PHBVHx), alone or in mixtures. [Claim 89]. Biomaterial according to any one of the preceding claims, characterized in that the PHA is the P4HB. [Claim 90]. Biomaterial according to any one of the preceding claims, characterized in that PHA is poly(3-hydroxybutyrate-co-4-hydroxybutyrate). [Claim 91]. Biomaterial according to any one of the preceding claims, characterized in that PHA is P4HB in mixture with poly(3-hydroxybutyrate-co-4-hydroxybutyrate. [Claim 92]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particles have a size strictly less than 100 pm. [Claim 93]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particles are prepared by microfluidization. [Claim 94]. Biomaterial according to any one of the preceding claims, characterized in that the PHA particles are prepared by a membrane emulsification process. [Claim 95]. Biomaterial according to any one of the preceding claims, characterized in that the composition further comprises an active ingredient selected from the group consisting of local anesthetics, vitamin C derivatives, anti-inflammatories, antioxidants, antibiotics, bacteriostatics, antihemorrhagics or hemostatics, and mixtures thereof. [Claim 96]. Biomaterial according to any one of the preceding claims, characterized in that the composition further comprises at least one active ingredient selected from the group consisting of antibiotics, bacteriostatics, alone or in mixture. [Claim 97]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one antibiotic, alone or in a mixture. [Claim 98]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one antibiotic selected, for example, from the group of rifampicin, minocycline, gentamicin, fusidic acid, amoxicillin, azithromycin, benzathine, benzylpenicillin, cefaclor, cefadroxil, cephalexin, cefixime, cefotiam, cefpodoxime, ceftriaxone, cefuroxime, ciprofloxacin, clarithromycin, clindamycin, cloxacillin, cotrimoxazole, doxycycline, erythromycin, fosfomycin, josamycin, levofloxacin, metronidazole, minocycline, moxifloxacin, mupirocin, nitrofurantoin, norfloxacin, ofloxacin, ornidazole, phenoxymethylpenicillin (penicillin V), penicillin, pivmecillinam, pristinamycin, roxithromycin, spiramycin, sulfadiazine, tetracycline and trimethoprim, alone or in combination. [Claim 99]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one mixture of two antibiotics, such as a mixture of rifampicin and minocycline, or of gentamicin and minocycline. [Claim 100]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one bacteriostatic agent, alone or in a mixture. [Claim 101]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one antihemorrhagic or hemostatic agent. [Claim 102]. Biomaterials according to any one of the preceding claims, characterized in that the composition further comprises at least one antihemorrhagic or hemostatic agent selected from the group comprising aminocaproic acid or tranexamic acid, alone or in mixture.