WO2024142055A1 - Bioadhesive microneedles array and use thereof - Google Patents

Abstract

A polymeric article comprising a plurality of convergent vertical structures bound to a support, wherein the article is composed of (i) a non-crosslinked biodegradable polymer, (ii) a crosslinked polymer, and (iii) a tissue-adhesive polymer, and use thereof such as for sealing a biological tissue is provided.

Description

BIOADHESIVE MICRONEEDLES ARRAY AND USE THEREOF

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of U.S. Provisional Patent Application No. 63/435,453, filed December 27, 2022, entitled “BIOADHESIVE MICRONEEDLES ARRAY AND USE THEREOF”, the contents of which are incorporated herein by reference in their entirety.

FIELD OF THE INVENTION

[0002] The invention relates generally to the field of medical devices comprising biodegradable adhesive arrays and methods for use thereof such as for sealing a tissue or for delivery of an active agent to a subject in need thereof.

BACKGROUND

[0003] Adhesion of soft tissues, as well as their sealing, are of great significance in the clinical and medical fields. Although various formulations have been developed for soft- tissue adhesives, tradeoffs between adhesive properties and tissue toxicity limit their clinical use. Cyanoacrylate derivatives, for example, adhere very strongly to tissue, but are very toxic. In contrast, fibrin sealants and other hydrogel adhesives generally have excellent biocompatibility but adhere very weakly to tissues.

[0004] Microneedles (MN) are minimally invasive device composed of micron-sized needles arranged on a small patch. MNs has several key advantages, such as minimal invasiveness, pain-free penetration, simple administration and better compliance compared to alternative treatments such as standard needles.

[0005] To this end, there is an unmet need for the development of tough bio-adhesive and bio-compatible MNs composed of biodegradable materials, for use in the biomedical and pharmaceutical fields such as inducing or facilitating healing of a damaged tissue. [0006] The foregoing examples of the related art and limitations related therewith are intended to be illustrative and not exclusive. Other limitations of the related art will become apparent to those of skill in the art upon a reading of the specification and a study of the figures.

SUMMARY

[0007] The following embodiments and aspects thereof are described and illustrated in conjunction with systems, tools and methods which are meant to be exemplary and illustrative, not limiting in scope.

[0008] In one aspect of the invention, there is provided an article comprising a support and a plurality of vertically aligned structures bound to the support, wherein the article is composed essentially of biocompatible constituents and comprises (i) a crosslinked polymer, and (ii) a tissue-adhesive polymer; the article is characterized by a retention on or within a biological tissue; each convergent structure of the plurality of vertically aligned structures is characterized by (i) a force at failure of at least 0.5 N, and (ii) swellability of at least 100%; and an average length of the plurality of vertically aligned structures is between 0.1 and 2 mm; and wherein the support and the plurality of vertically aligned structures are composed of different polymers.

[0009] In some embodiments, the support comprises the tissue adhesive polymer.

[0010] In some embodiments, each of the plurality of vertically aligned structures is in a form of convergent structure; and wherein the plurality of vertically aligned structures are arranged in an array on top of the support.

[0011] In some embodiments, the plurality of vertically aligned structures comprises the crosslinked polymer.

[0012] In some embodiments, a density of the plurality of vertically aligned structures within the array is between 20 and 1000 units/cm2.

[0013] In some embodiments, a center-to-center distance between a pair of adjacent vertically aligned structures within the array is between 10 and 2000 pm. [0014] In some embodiments, the article is characterized by a force at failure of at least 0.2N force/needle.

[0015] In some embodiments, the array is characterized by a force at failure of between 4 and 200N.

[0016] In some embodiments, the article is characterized by sealing capability of at least 120 mmHg.

[0017] In some embodiments, each of the plurality of vertically aligned structures has a pyramidal shape.

[0018] In some embodiments, each of the plurality of vertically aligned structures comprises a base portion in contact with the support, and a top portion distal to the base portion.

[0019] In some embodiments, an average width dimension of the base portion the dimension is between 50 and 1000 pm.

[0020] In some embodiments, an average width dimension of the top portion is between 0.1 and 10 pm.

[0021] In some embodiments, the crosslinked polymer comprises an acrylate -based polymer.

[0022] In some embodiments, the tissue-adhesive polymer comprises alginate, chitosan, pectin, hyaluronic acid, derivates of PEG, cellulose, cellulose derivate, Collagen, Fibrinogen, and polyacrylate, including any salt, any derivative, any copolymer, or any combination thereof.

[0023] In some embodiments, the support is further in contact with a capping layer.

[0024] In some embodiments, the article of the inventions, further comprising a pharmaceutically active ingredient.

[0025] In some embodiments, the support consists essentially of chitosan, and wherein the plurality of vertically aligned structures consists essentially of crosslinked polyacrylate. [0026] In another aspect, there is provided a method for preventing or treating a medical condition in a subject in need thereof, comprising administering the article of the invention to a subject, thereby preventing or treating the medical condition.

[0027] In some embodiments, the medical condition comprises a damage of a biological tissue of the subject.

[0028] In some embodiments, the administering comprises contacting the article with a biological tissue of the subject.

[0029] In some embodiments, the biological tissue comprises a mucosal tissue, a dermal tissue, a muscle tissue, or any combination thereof.

[0030] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the figures and by study of the following detailed description.

[0031] Exemplary embodiments are illustrated in referenced figures. Dimensions of components and features shown in the figures are generally chosen for convenience and clarity of presentation and are not necessarily shown to scale. The figures are listed below:

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figures 1A-1D are schemes presenting polydimethylsilxane (PDMS) mold manufacture process. Figure 1A. The master template parts. Figure IB. The PDMS solution will be poured into the mold (marked with an arrow). Figure 1C. The solution inside the closed mold. Figure ID. Separation of the PDMS mold from the master mold.

[0033] Figure 2 is a scheme presenting microneedle patch manufacturing process using micromolding method.

[0034] Figures 3A-3B are graphs and images presenting mechanical properties of different polymeric MN arrays versus commercial controls. Figure 3A. representative forceextension curves of different polymeric MN arrays. Pullulan (*), PLGA (**), Carbopol (***), Chitosan (****), Carbopol/Chitosan (*****). [0035] Figure 3B. Needle failure of different polymeric MN arrays. Data are means ± SD, n = 4. * = p < 0.05, *** = p < 0.005.

[0036] Figures 3C1-3C’5. SEM images before (3C1-3C5) and after (3C’ 1-3C’5) compression of different polymeric MN arrays. PLGA (Cl/C’ l), Pullulan (C2/C’2), Carbopol (C3/C’3), Chitosan (C4/C’4), Carbopol/Chitosan (C5/C’5).

[0037] Figure 3D is a bar graph presenting burst pressure results for polymeric MN arrays versus commercial controls Evicel® and Duraseal®.

[0038] Figure 4 is a graph presenting PBS swelling and erosion of tested MNs sealants and commercial Evicel® (pH 7.4) at 37 °C.

[0039] Figures 5A-5B are graphs and images presenting cytotoxicity of the tested MNs sealants versus commercial controls. Figure 5A. NIH 3T3 cell viability compared with unexposed cells, evaluated by MTS assay 48 h after exposure to different polymeric MN patches and to commercial octyl-cyanoacrylate (Dermabond®), Evicel® and Surgicel® patches. Figure 5B. Evaluation of cell viability by Live/dead assay. Data are means ± SD, n = 4. *** = p < 0.005.

[0040] Figures 6A-6C. Fig. 6A: Schematic illustration of the incision procedure and the MN patch treatment on rat liver. Fig. 6B: Incision and treatment of rat liver bleeding model without any treatment (control), with MN patch, and with Surgicel®, respectively (scale bar 1 cm). Fig. 6C: Blood loss for the three treatment groups (n = 4, *p < 0.05, **p < 0.01).

DETAILED DESCRIPTION

[0041] Disclosed herein are articles (e.g. medical devices) and a method for applying thereof to a target site of a subject, wherein the article is characterized by an adhesiveness and swellability at the target site, and wherein the article is at least partially biodegradable. In some embodiments, the present article is characterized by muco-adhesiveness, sufficient toughness and swellability when applied to a tissue (e.g. a skin tissue). In some embodiments, the present article is particularly useful for tissue sealing and/or healing of a damaged tissue.

[0042] The inventors developed an article in a form of a polymeric patch with a needlelike array capable of penetrating a soft tissue (e.g. a skin tissue), and subsequently interlocking upon swelling. The inventors postulate, that the sharp tip of the needle-like structure enables the needle-like structures to penetrate a tissue and upon water absorption the needle-like structures undergo swelling, so as to affix the article to the soft tissue via physical interlocking. The polymer material of the article of the invention is characterized by sufficient elasticity and toughness and is configured to retain at the application site, so as to allow sealing and to promote healing of a damaged tissue. The present invention, in some embodiments thereof, is based in a surprising finding that a polymeric material comprising a combination of a mucoadhesive polymer (e.g. chitosan) with a cross-linked biodegradable polymer (.e.g. PLA), and further comprising a non-crosslinked biodegradable polymer resulted in superior toughness and swellability, allowing application of the article of the invention to a soft tissue (e.g. skin).

Article

[0043] According to one aspect of the present invention, there is an article comprising a support and a plurality vertically aligned structures bound to the support, wherein the article comprises a crosslinked polymer, and/or a tissue adhesive polymer. In some embodiments, the article comprises a plurality of polymers.

[0044] According to another aspect of the present invention, there is an article comprising a support and a plurality vertically aligned structures bound to the support, wherein the article comprises a first polymer and a second polymer. In some embodiments, the first polymer and the second polymer are chemically distinct polymers. In some embodiments, the first polymer comprises a crosslinked polymer, and the second polymer comprises a tissue adhesive polymer. In some embodiments, the support and the plurality of vertically aligned structures of the article are composed of different polymers. [0045] As used herein, the terms “first polymer” and “crosslinked polymer” are used herein interchangeably. As used herein, the terms “second polymer” and “tissue-adhesive polymer” are used herein interchangeably.

[0046] As used herein the term “adhesive” refers to adhesiveness of the article or a part thereof to a biological tissue. In some embodiments, the term “adhesiveness” or the term “sufficient adhesiveness” (which are used herein interchangeably) refers to the ability of the film and/or article of the invention to form a stable bond (e.g., non-covalent, or physical interaction) to at least one surface of the biological tissue. Thus, term adhesiveness is well known in the art, an may be related to physical forces (bonds or interactions, including dipole-dipole interactions, hydrogen bonds, London forces, Van-der-Waals forces, or any physical forces such as vacuum, capillary forces, etc.) that exists in the area of contact between two surfaces (e.g. the outer surface of the film of the invention and the outer surface of the biological tissue), holding them together. As used herein, the term "biological tissue" refers to any surface comprising cells and/or biological molecules (e.g. proteins, polysaccharides, lipids, nucleic acids).

[0047] In some embodiments, any one of the first polymer and the second polymer is a biocompatible polymer. In some embodiments, at least one of the first polymer and the second polymer is a biodegradable polymer and a biocompatible polymer.

[0048] In some embodiments, the article of the invention comprises a bottom polymeric layer, referred to as the “support”, comprising or in contact with an array of vertically aligned structures. In some embodiments, the vertically aligned structures are convergent (or needle- shaped) structures.

[0049] In some embodiments, the support is characterized by tissue adhesiveness and/or sealing capability, as disclosed hereinbelow. In some embodiments, the support comprises a tissue-adhesive polymer (the second polymer). In some embodiments, the support comprises a tissue-adhesive polymer and at least one of: the crosslinked polymer and the non-crosslinked biodegradable polymer, including any combination thereof. In some embodiments, the support comprises, a crosslinked polymer e.g. and a tissue-adhesive polymer (e.g. e.g. chitosan). In some embodiments, the support comprises e.g. a tissueadhesive polymer (e.g. cellulose).

[0050] In some embodiments, the support is a homogenous polymeric film. In some embodiments, the film comprises a polymeric matrix. In some embodiments, the film comprises a polymeric biodegradable matrix. In some embodiments, the film comprises a polymeric adhesive matrix. In some embodiments, the film comprises a polymeric mucoadhesive matrix. In some embodiments, the film comprises a polymeric biodegradable mucoadhesive matrix.

[0051] In some embodiments, the film is a single layer. In some embodiments the film, is a multi layer. In some embodiments, the film layer, is in a form of a continuous layer.

[0052] The term “continuous layer” or the term “layer” refers to a substantially homogeneous substance of substantially uniform-thickness which maintains its physicochemical properties (e.g. mechanical strength, elasticity, Young’s modulus, chemical composition, water content) with the entire dimensions (lengths and width dimensions) thereof. In some embodiments, each layer has a different physical structure and/or a different chemical composition. In some embodiments, each layer has the same physical structure and/or the same chemical composition. In some embodiments, the term "layer", refers to a protein layer.

[0053] In some embodiments, the film comprises one or more layer(s), wherein each of the one or more layer(s) is characterized by a thickness between 10 pm and 500 pm, between 100 pm and 1000 pm, between 1 mm and 10 mm, between 2 mm and 5mm, between 5 pm and 200 pm, between 1 mm and 5mm, including any range or value therebetween. Each possibility represents a separate embodiment of the invention.

[0054] The term “thickness” refers to the dry thickness. As used herein, the term “dry thickness” refers to the thickness of the dried film layer (e.g. upon substantial evaporation or removal of water). Dried film layer refers to a film layer in a solid state (e.g. non-flowable layer, substantially retaining its shape and/or dimensions upon tilting thereof). In some embodiments, the terms “thick” or “thickness” including any grammatical form thereof, refer to an average thickness.

[0055] In some embodiment, the entire support has the same chemical composition. In some embodiment, the entire support is composed of the same polymers. In some embodiment, the support comprises of: a first adhesive surface and a second surface. In some embodiments, the second surface is substantially non-adhesive. In some embodiments, the second surface is devoid of adhesiveness.

[0056] In some embodiments, the support is in a form of a film in contact with or bound to the vertically aligned structures. In some embodiments, the film comprises the polymeric layer in contact with the vertically aligned structures. In some embodiments, the vertically aligned structures are stably bound to the polymeric layer. In some embodiments, the vertically aligned structures and the polymeric film are fused or molten together. In some embodiments, the vertically aligned structures are joined with the polymeric layer. In some embodiments, the polymeric layer has a first outer portion facing the vertically aligned structures and a second outer portion opposed to the first outer portion, wherein the vertically aligned structures are stably bound or linked to the first outer portion of the polymeric layer.

[0057] In some embodiments, bound is via a non-covalent bond (e.g. electrostatic interaction, van-der-Waals bond, dipole-dipole interactions, hydrogen bond, London forces or any combination thereof). In some embodiments, bound is via a covalent bond. The terms “covalent bond” and “non-covalent bond” are well-understood by a skilled artisan.

[0058] In some embodiments, the plurality of vertically aligned structures comprises or consist essentially of one or more swellable polymer(s). In some embodiments, the plurality of vertically aligned structures consists essentially of one or more swellable biocompatible polymer(s). In some embodiments, each of the plurality of vertically aligned structures consist essentially of a homogenous polymeric material. In some embodiments, each of the plurality of vertically aligned structures and the support is substantially devoid of a particulate matter. [0059] The term “swellability” refers to the ability of the plurality of vertically aligned structures of the invention (and/or a portion thereof) to absorb water and as a consequence to increase its volume (i.e. to expand). In some embodiments, water molecules are bonded by physical interactions (e.g. hydrogen bonding, dipol-dipol interactions, electrostatic interactions, etc.) to the one or more polymer(s) composing the vertically aligned structures. In some embodiments, the term “swellability” refers to a weight (or volume) increase of the article due to water absorption, relative to the initial weigh (or volume) of the article.

[0060] In some embodiments, the support comprises or consists essentially of a biodegradable polymer. In some embodiments, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the support consists of one or more tissue adhesive polymer(s). In some embodiments, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the support consists of a single tissue adhesive polymer.

[0061] In some embodiments, the second outer portion of the support is further in contact with a capping layer (i.e. a barrier layer composed of a non-adhesive polymer).

[0062] In some embodiments, the plurality of vertically aligned structures comprises or consist essentially of a single polymer species or a plurality of chemically distinct polymers.

[0063] The term “biocompatible” refers to polymers that are generally considered as being non immunogenic and are considered as being non-toxic or harmful to a subject upon exposure thereto. In some embodiments, the term biocompatible refers to polymers approved by a regulatory authority such as FDA.

[0064] The term “biodegradable” describes a substance which can decompose under environmental condi tion(s) into breakdown products. In some embodiments, the term "biodegradable" as used in the context of embodiments of the invention, also encompasses the term "bioerodible", which describes a material/ which decomposes under environmental conditions into smaller fractions, thus substantially losing its structure and/or mechanical properties. In some embodiments, the term “bioerosion” refers to erosion of the polymeric material initiated by water (e.g. by dissolution), enzymes, etc., and resulting in at least partial degradation of the composition/article comprising the bioerodible material.

[0065] In some embodiments, the crosslinked polymer consist essentially of a polymer and a crosslinker; wherein the polymer comprises any one of: acrylate -based polymer (e.g., polyacrylic acid, poly(methacrylate), poly(ethacrylate), poly(methyl acrylate), poly(ethyl acrylate)), polyvinyl alcohol (PVA), cellulose, and cellulose derivatives, including any copolymer thereof, and/or any combination thereof (i.e. polymer blend). In some embodiments, the crosslinked polymer consist essentially of a single polymer species and a single crosslinker species. In some embodiments, the crosslinked polymer further comprises an additional non-crosslinked biodegradable/biocompatible polymer (e.g. PVP, PEG, a polyester, acrylate-based polymer, starch, cellulose, and/or a cellulose derivative). In some embodiments, the additional non-crosslinked biodegradable/biocompatible polymer is or comprises PVP-iodine (povidone-iodine).

[0066] In some embodiments, the cellulose derivative comprises any of: alkylated cellulose (e.g. ethyl cellulose, methyl cellulose, etc.), carboxylated cellulose (e.g. carboxymethyl cellulose (CMC), etc.), hydroxylated cellulose (e.g. hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC) etc.).

[0067] In some embodiments, the derivative of cellulose is a hydroxylated cellulose comprising a repeating unit represented by Formula 1 :

wherein each R independently represents H or a hydroxyalkyl.

[0068] In some embodiments, the derivative of cellulose is a carboxylated cellulose comprising a repeating unit represented by Formula 1, wherein each R independently represents alkyl-carboxy, or H. In some embodiments, the derivative of cellulose is an alkylated cellulose comprising a repeating unit represented by Formula 1, wherein R independently represents an alkyl or H.

[0069] In some embodiments, at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the crosslinked polymer consist of the polymer disclosed above and the crosslinker.

[0070] The term “cross-linking” as used herein refers to the formation of a chemical bond or a physical interaction (e.g. ionic bond) between two chemical moieties or groups. In some embodiments, cross-linked comprises inter cross-linked (e.g. wherein the chemical moieties are distinct polymeric chains). In some embodiments, cross-linked comprises intra crosslinked (e.g. wherein the chemical moieties are within the same polymeric chain).

[0071] In some embodiments, the crosslinker is a covalent crosslinker. In some embodiments, the crosslinker is a multi-functional crosslinker.

[0072] A skilled artisan will appreciate that various crosslinkers may be implemented herein, depending on the nature of the polymer. For example, PVA or cellulose may be crosslinked via crosslinkers comprising two or more carboxy groups bound to an alkyl backbone, alternatively polyacrylic acid can be crosslinked via a crosslinker comprising two or more hydroxy groups bound to an alkyl backbone.

[0073] In some embodiments, the crosslinking degree of the crosslinked polymer (i.e. a weight portion of the crosslinker relative to the entire weight of the crosslinked polymer) is between 1 and 40%, between 1 and 30%, between 1 and 20%, between 1 and 10%, between 1 and 5%, between 10 and 40%, between 10 and 30%, between 1 and 20%, between 1 and 10%, between 1 and 5%, including any range between.

[0074] In some embodiments, an average MW of the crosslinked polymer is between 100.000 and 3.000.000 Da, between 500.000 and 3.000.000 Da, between 100.000 and 2.000.000 Da, between 100.000 and 1.000.000 Da, between 500.000 and 2.000.000 Da, including any range between. [0075] In some embodiments, the crosslinked polymer is crosslinked polyacrylic acid. In some embodiments, the crosslinked polymer is a cross-linked polyacrylate, including any co-polymer and any salt thereof.

[0076] In some embodiments, the crosslinked polymer is polyacrylic acid crosslinked via a crosslinker composing Cl -CIO alkyl (linear or branched) substituted by a plurality (e.g. 2, 3, 4, 5, 6) of hydroxy, amino, or mercapto groups. In some embodiments, the crosslinker is C2-C10-diol (such as 1,5 -pentanediol, 1,6-hexanediol, 1,4-propanediol), glycerol, pentaerythritol, dipentaerythritol, or tripentaerythritol, including any combination thereof.

[0077] In some embodiments, the crosslinked polymer is polyacrylic acid crosslinked by pentaerythritol, such as Carbopol 941.

[0078] In some embodiments, the w/w percentage of the crosslinked polymer in the article of the invention ranges between 10 and 50%, between 10 and 20%, between 10 and 15%, between 20 and 30%, between 30 and 40%, between 40 and 50% and between 15 and 20% including any range between.

[0079] In some embodiments, the weight percentage of the tissue-adhesive polymer in the article of the invention ranges between 10 and 50%, between 10 and 30%, between 15 and 35%, between 20 and 40%, between 40 and 50%, and between 25 and 35% including any range between.

[0080] In some embodiments, the article of the invention consists essentially of the tissueadhesive polymer, the crosslinked polymer, and optionally a pharmaceutically active ingredient.

[0081 ] In some embodiment, the tissue-adhesive polymer of the invention comprises any one of: a tissue adhesive derivative of cellulose (e.g. cellulose ester, hydroxylated cellulose, alkylated cellulose, such as HPMC etc.), pectin, alginate, PED derivatives, hyaluronic acid, pullulan, fibrinogen, collagen, chitosan and non-crosslinked polyacrylate (or acrylate polymer), including any salt, any derivative, any copolymer, or any combination thereof. In some embodiments, the tissue-adhesive polymer is a tissue-adhesive polysaccharide. In some embodiments, the tissue-adhesive polymer is a mucoadhesive polymer. In some embodiments, the tissue-adhesive polymer is selected from chitosan and carboxylated cellulose, including any blend or any copolymer thereof.

[0082] In some embodiment, the tissue-adhesive polymer is a tissue-adhesive polysaccharide (i.e. a non-crosslinked polysaccharide). In some embodiment, the tissue adhesive polysaccharide comprises chitosan, pectin, alginic acid, hyaluronic acid, tissue adhesive cellulose derivative; including any salt, any derivative, any copolymer, or any combination thereof. In some embodiment, the tissue-adhesive polysaccharide comprises or consists essentially of a natural polysaccharide, or a modified polysaccharide (including side-chain modification such as alkylation or additional modifications described for the cellulose derivatives above, de-acetylation, etc.).

[0083] In some embodiment, the tissue-adhesive polymer is characterized by an average MW between 50.000 and 3.000.000 Da, between 50.000 and 2.000.000 Da, between 100.000 and 1.000.000 Da, between 100.000 and 1.000.000 Da, between 100.000 and 500.000 Da, between 100.000 and 400.000 Da, between 100.000 and 350.000 Da, including any range between.

[0084] In some embodiments, the w/w ratio between the crosslinked polymer, and the tissue adhesive polymer within the article of the innovation is between 1: 1 and 4: 1, between 1: 1 and 2: 1, between 1:1 and 3:1 and between 1: 1 3.5:1 including any range between.

[0085] In some embodiment, the plurality of vertically aligned structures is composed essentially of a crosslinked polyacrylate (e.g. Carbopol) including any salt and any copolymer thereof, and the support is composed essentially of the tissue adhesive polymer (e.g. chitosan) including any salt and any copolymer thereof.

[0086] In some embodiments, at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the plurality of vertically aligned structures consist of the crosslinked polyacrylate.

[0087] In some embodiments, at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the support consist of the tissue-adhesive polymer; and at least 90%, at least 95%, at least 99%, at least 99.9%, between 70 and 99.9%, between 70 and 90%, between 80 and 99.9%, including any range between by weight of the plurality of vertically aligned structures consist of the crosslinked polymer.

[0088] In some embodiment, the plurality of vertically aligned structures is composed essentially of a crosslinked polyacrylate (e.g. Carbopol) including any salt and any copolymer thereof, and the support is composed essentially of chitosan including any salt and any copolymer thereof.

[0089] In some embodiments, the chitosan is a medium molecular weight chitosan (MMWC), characterized by an average MW between 180 and 350KDa and by a deacetylation degree between 70 and 99%, or between 70 and 90%, or between 75 and 90%, or between 75 and 85%, including any range between.

[0090] In some embodiments, the plurality of vertically aligned structures is in a form of an array. In some embodiments, the array is in a form of a single row, or a plurality of rows, which may be parallel. In some embodiments, the array is linear or curved. In some embodiments, the plurality of vertically aligned structures is arranged on the polymeric layer in a form of two or more parallel rows, so as to form parallel arrays.

[0091] In some embodiments, the vertically aligned structures are arrayed in one or more of a circular array, an oval array, a rectangular array, a triangular array, a polygon array, a columnar array, and/or a horseshoe array. In some embodiments, the array defines an axis (e.g. a longitudinal axis) of the substrate. In some embodiments, the vertically aligned structures are arranged along an axis (i.e. have a directional orientation) on top of the support.

[0092] In some embodiments, each of the vertically aligned structures is characterized by a force at failure of at least about 0.05N, at least about 0.1N, at least about 0.2N, at least about IN, between 0.05 and 0.5N, between 0.05 and 0.1N, including any range or value therebetween (as determined by a texture analyzer). In some embodiments, the force at failure refers to an average force per single structure. [0093] In some embodiments, the array of the vertically aligned convergent structures is characterized by an average force at failure of at least 4N, at least 7N, at least ION, at least 20N, at least 50N, at lease 100N, at least 150N, at least 200N and between 4 and 50N, between 50 and 100N, between 100 and 150N, between 150 and 200N, including any range or value therebetween.

[0094] In some embodiments, the vertically aligned structures are characterized by a swellability of at least 100%, at least 120% at least 150%, at least 200%, at least 300% w/w, at least 500% w/w, at least 1000%, at least 2000%, at least 3000%, at least 4000%, at least 4500%, at least 500%, between 500 and 5000%, between 900 and 5000%, between 1000 and 5000%, between 2000 and 5000%, including any range or value therebetween.

[0095] In some embodiments, the average length of vertically aligned structures is between 0.01 and 2 mm, between 2 and 5 mm, between 0.01 and 0.1 mm, between 0.1 and 1 mm, between 1 and 2 mm and between 0.5 and 1mm, including any range or value therebetween.

[0096] In some embodiments, the density of the vertically aligned structures within the array is between 20 and 1000 units/cm², between 20 and 1000 units /cm², between 500 and 2000 units/cm², between 20 and 500 units/cm², between 200 and 1000 units/cm², between 800 and 1000 units/cm², and between 500 and 800 units/cm², including any range or value therebetween.

[0097] In some embodiments, the term “density” refers to a number of vertically aligned structures per surface area of the film (e.g. within a square centimeter). In some embodiments, the density refers to a number of vertically aligned structures per surface area of the array. One skilled in the art will appreciate, that if the vertical structures are uniformly distributed on or within the film of the invention (e.g. on top of the polymeric layer), the density within the entire surface of the film is substantially the same. However, if the vertical structures are non-uniformly distributed on or within the film surface (such as forming distinct arrays of vertical structures), the density refers to the number of vertically aligned structures per surface area of the array. [0098] In some embodiments, the vertically aligned structures are characterized by a random geometric form or shape. In some embodiments, the vertically aligned structures are characterized by a predefined geometric form or shape. In some embodiments, the vertically aligned structures are uniformly shaped. In some embodiments, a tri-dimensional shape of at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99% of the vertically aligned structures is the same or different.

[0099] In some embodiments, the vertically aligned structures have a convergent shape, wherein each of plurality vertically aligned convergent structures comprises a base portion in contact with the support, and a top portion distal to the base portion, and wherein the cross-section of the base portion is greater than the cross-section of the top portion. In some embodiments, the cross-section of the base portion is greater than the cross-section of the top portion by at least 2, at least 5, at least 10 times, between 2 and 10 times, between 2 and 100 time, between 10 and 100 times, including any range between.

[0100] In some embodiments, the vertically aligned structures are as described herein and have a pyramidal shape (e.g. having a shape of a tetrahedron, a pentagonal pyramid, a hexagonal pyramid etc.), a conical shape, or are irregularly shaped.

[0101] In some embodiments , a ratio between the height of the vertically aligned pyramidal shaped structures and their base is between 2: 1 and 10: 1, between 2: 1 and 3:1, between 3:1 and 4:1, between 4: 1 and 5: 1, between 2: 1 and 5: 1, including any range therebetween.

[01 2] In some embodiments, the average width dimension of the base portion is between 50 and lOOOp, between 50 and 300p, between 50 and 500p, between 200 and 500p, between 50 and 750p, between 100 and lOOOp, between 100 and 500p, between 100 and 300p, including any range or value therebetween.

[01 3] In some embodiments, the average width dimension of the top portion is between 0.1 and lOp, between 1 and lOOOnm, between 1 and lOnm, between 5 and lOOnm, between 10 and 200 nm, and between 20 and 500nm, including any range or value therebetween.

[0104] In some embodiments, the center-to-center distance between a pair of adjacent vertically aligned structures within the array is between 10 and 2000 pm, between 10-200 pm, between 100 and 500 pm, between 250 and 500 pm, between 500 and 750 pm, between 1000 and 1500 pm and between 1500 and 2000 pm, including any range or value therebetween.

[0105] In some embodiments, a center-to-center distance refers to a distance between adjacent vertically aligned structures within the array.

[0106] As used herein, the term “vertically aligned structures” does not necessarily refer to perpendicular alignment of the structures. In some embodiments, the vertically aligned structures are substantially perpendicular to the plane of the polymeric layer 101, wherein substantially is as described herein. In some embodiments, the vertically aligned structures are lateral to the plane of the polymeric layer. In some embodiments, at least a portion of the vertically aligned structures has an angle of between 70 and 120°, between 70 and 80°, between 80 and 90°, between 90 and 100°, between 100 and 120° relative to the plane of the polymeric layer including any range therebetween.

[0107] In some embodiments, the vertically aligned structures are directly bound to the support (so that there is no additional intermediate layer between the support and the vertically aligned structures). In some embodiments, the base portion of the vertically aligned structures is in direct contact with the support.

[01 8] The number of vertically aligned structures within the article/or within a single array of the article may vary depending on dimensions of the article. In some embodiments, the number of vertically aligned structures within the article/single array is at least 5, at least 10, at least 20, at least 50, at least 100, at least 1000, at least 10.000, at least 50.000, including any range or value between.

[0109] In some embodiments, the article of the invention is biocompatible. In some embodiments, at least 80%, at least 90%, at least 95%, at least 99%, or between 90 and 100% by dry weight of the article is composed of biocompatible constituents.

[0110] In some embodiments, the article of the invention is at least partially biodegradable. In some embodiments, the article of the invention comprises one or more biodegradable and one or more non-biodegradable polymer. In some embodiments, the support and the vertically aligned structures are characterized by the same degradation rate. In some embodiments, the polymeric layer and the vertically aligned structures are characterized by different degradation rates. One skilled in the art will appreciate, that the degradation rate may be predetermined by the chemical composition, and/or by the geometrical shape or dimension of the polymeric layer and of the vertically aligned structures.

[0111] In some embodiments, a w/w concentration of the non-degradable polymer within the article of the invention is between 0.1 and 60%, between 0.1 and 1%, between 1 and 5%, between 5 and 10%, between 0.1 and 10%, between 0.1 and 20%, between 10 and 20%, between 20 and 40%, between 40 and 60%, including any range therebetween.

[0112] In some embodiments, the article of the invention comprises a non -biodegradable polymer. In some embodiments, the terms “non-degradable” and “non-biodegradable” are used herein interchangeably.

[01 13] In some embodiments, the degradation rate of the article is predetermined by the chemical composition of the polymers composing thereof, average molecular weight (MW), and/or the structure (e.g. branched versus linear polymer; block co-polymer versus graft copolymer; number of blocks and/or MW of any one of the blocks) of the polymers composing thereof. In some embodiments, the article thickness of the invention ranges between 100 pm and 10mm, including any range or value therebetween.

[0114] In some embodiments, the article of the invention is configured to be stably bound to an application site. In some embodiments, the article of the invention is substantially retained at the application site for at least Ih, at least 2h, at least O.lh, at least 5h, at least 10 h, at least 24h, at least 48 h, or between 0.1 and 48h, between 1 and 48h, including any range between. In some embodiments, the application site is a predetermined location on or within a biological tissue. In some embodiments, the biological tissue is a soft tissue. In some embodiments, the biological tissue comprises one or more of: a dermal tissue, a mucosal tissue, a muscle tissue, or any combination thereof. In some embodiments, the biological tissue comprises a damaged tissue (i.e. a tissue containing a wound, injury, burn, incision, necrosis, bleeding, or leakage of a biological fluid). [0115] In some embodiments, the retention of the article of the invention at the application site is predetermined by the swelling of the vertical structures. In some embodiments, the vertical structures are characterized by a swellability sufficient for retaining the article at the application site. In some embodiments, sufficient swellability of the vertical structures is as described hereinbelow.

[0116] In some embodiments, the support of the invention is characterized by an inner surface facing and/or in contact with the vertically aligned structures, and by an outer surface facing the ambient. In some embodiments, the inner surface is characterized by adhesiveness to the biological tissue. In some embodiments, the inner surface is devoid of adhesiveness to the biological tissue.

[01 17] In some embodiments, the swellability of the article of the invention ranges between 80% and 1500%, between 50 and 200%, between 200 and 400%, between 400 and 600%, between 700 and 900%, between 800 and 1250%, including any range or value therebetween.

[0118] In some embodiments, the support of the article of the invention is characterized by a swellability ranging between 0% and 10%, between 10 and 20%, between 5 and 15%, between 15 and 50%, between 10 and 15%, between 15 and 20%, between 1 and 50% including any range or value therebetween.

[0119] In some embodiments, the vertical structures of the article of the invention are characterized by a swellability ranging between 100% and 7000%, between 100 and 6000%, between 200 and 5000%, between 500% and 5000%, between 1000 and 5000%, between 2000 and 4500%, between 2000 and 4500%, between 700 and 900%, between 800 and 2000%, between 400 and 600%, between 700 and 900%, between 1000 and 4500%, including any range or value therebetween.

[0120] The term “swellability” refers to the ability of the article of the invention (and/or a portion thereof) to absorb water and as a consequence to increase its volume (i.e. to expand). In some embodiments, water molecules are bonded by physical interactions (e.g. hydrogen bonding, dipol-dipol interactions, electrostatic interactions, etc.) to the one or more polymer(s) composing the vertically aligned structures. In some embodiments, the term “swellability” refers to a weight (or volume) increase of the article due to water absorption, relative to the initial weigh (or volume) of the article.

[0121] In some embodiments, the article of the invention is characterized by a tissue sealing capability. The sealing capability of the article of the invention is sufficient to withstand blood pressure in the blood vessel, which is typically 120 mmHg. In some embodiment, the article of the invention (or the support of the article, as disclosed herein) is characterized by a burst pressure of at least about 100 mmHg, at least 120mmHg, and least 150 mmHg, and between 100 and 500 mmHg, between 200 and 500 mmHg, between 100 and 300 mmHg, between 100 and 700 mmHg, between 100 and 1000 mmHg, including any range or value therebetween (as measured by ASTM F2392-04).

[0122] In some embodiments, the article is characterized by a greater sealing capability as compared to a commercial product (e.g. fibrin glue). In some embodiments, the article is characterized by a greater sealing capability of at least 2 times greater, at least 5, at least 10, at least 15, and at least 20 times more compared to a commercial fibrine glue, including any value therebetween.

[0123] In some embodiments, the article of the invention is characterized by a tensile strength of at least 10 MPa and between 10 and lOOOMPa, between 10 and lOOMPa, between 10 and 500 MPa, between 200 and 700MPa, between 200 and 500 MPa, between 500 and 700 MPa, between 700 and 1000 MPa, including any range between.

[0124] In some embodiments, the adhesive surface of the film is characterized by a water contact angle of at least 10°, at least 30°, at least 50°, at least 70°, at least 80°, including any range therebetween.

[0125] In some embodiments, the article of the invention is characterized by a Young’s modulus of at least 1 MPa, at least 10 MPa, at least 50 MPa, at least 100 MPa, at least IGPa including any range between.

[0126] In some embodiments, the article of the invention is characterized by elongation at break of at least 10% and between 10 and 700%, between 10 and 100%, between 10 and 200%, between 100 and 300%, between 200 and 400%, between 300 and 500%, between 400 and 600% and between 500 and 700%, including any range between.

[0127] In some embodiments, the article of the invention is characterized by adhesive strength (determined as disclose din the Examples section) of at least lOKpa, at least 30Kpa, at least 40Kpa, at least 50Kpa, between 40 and about 60Kpa, between 35 and about 60Kpa, between 40 and about 70Kpa, including any range between.

[0128] In some embodiments, the article is in a form of a patch. In some embodiments, the article is a tissue sealant. In some embodiments, the article is a wound sealant. In some embodiments, the article is a patch for sealing wounds and/or arresting of bleeding.

[0129] In some embodiments, the article further comprises a pharmaceutically active agent (e.g., an anti-inflammatory drug, an antiseptic agent or disinfectant such as iodine or povidone iodine, chlorhexidine, hydrogen peroxide (HP), HP precursor, etc.) incorporated within the article.

[0130] In some embodiments, the article further comprises an additive (e.g. a coloring agent, a taste agent, etc.).

[0131] In some embodiments, the article further comprises an active agent. In some embodiments, the active agent comprises a pharmaceutically active ingredient (e.g. a drug). In some embodiments, the article comprises a pharmaceutically effective amount of the active ingredient. In some embodiments, at least a portion of the article comprises a pharmaceutically active ingredient. In some embodiments, the pharmaceutically active ingredient is in contact with the article. In some embodiments, the pharmaceutically active ingredient is bound to the article. In some embodiments, the pharmaceutically active ingredient is embedded within the film. In some embodiments, the pharmaceutically active ingredient is embedded within the vertically aligned structures. In some embodiments, the pharmaceutically active ingredient is homogenously mixed with the article. In some embodiments, the pharmaceutically active ingredient is homogenously dispersed within the article. In some embodiments, the pharmaceutically active ingredient is substantially located within the entire article. In some embodiments, the pharmaceutically active ingredient is located on top of the article. In some embodiments, the pharmaceutically active ingredient is incorporated on and/or within the article. In some embodiments, the pharmaceutically active ingredient and the article are in a form of a composite. In some embodiments, the pharmaceutically active ingredient is homogenously distributed within the article. In some embodiments, the pharmaceutically active ingredient is non-homogenously distributed within the article.

[0132] In some embodiments, the pharmaceutically active ingredient is in a form of a solid on and/or within the article. In some embodiments, the pharmaceutically active ingredient is in a form of an amorphous solid on and/or within the article. In some embodiments, the pharmaceutically active ingredient is in a form of a crystal on and/or within the article, wherein crystal includes any polymorphous form of the pharmaceutically active ingredient. In some embodiments, the pharmaceutically active ingredient is in a form of a gel within the article.

[0133] In some embodiments, the pharmaceutically active ingredient is substantially located on or within the polymeric layer (support) or the vertically aligned structures comprising the article including any combination thereof.

[0134] In some embodiments, the article of the invention is configured to substantially release the active agent. In some embodiments, the article of the invention is configured to substantially release the active agent under physiological conditions (e.g. a temperature of about 36C, a pH between 5 and 8, exposure to bodily fluids (e.g. saliva), and exposure to a tissue of a subject, such as a wet tissue). In some embodiments, the article of the invention is configured to substantially release the active agent upon contact thereof with a mucosal tissue.

[01 5] In some embodiments, the article is configured to encapsulate the active agent at a w/w concentration between 0.1 and 50%, is between 0.1 and 1%, is between 1 and 5%, is between 5 and 10%, is between 10 and 15%, is between 15 and 20%, is between 20 and 30%, is between 30 and 40%, is between 40 and 50% by total weight of the article, including any range therebetween. [0136] In some embodiments, the article provides for a flexible patch-like substrate configured for external or internal application at the application site. In some embodiments, the article provides for a flexible patch-like substrate configured for external or internal application at the application site. In some embodiments, the article is for the treatment of a damaged biological tissue the subject. In some embodiments, the article is for the delivery of an active agent (e.g. a drug) to the subject.

[0137] In some embodiments, the article of the invention is characterized by a mechanical stability.

[0138] As used herein the term “stability” refers to the capability of the film of the invention to maintain its structural and/or mechanical integrity. In some embodiments, the article is referred to as stable, if the article is characterized by a mechanical integrity sufficient to maintain the position and/or the vertical alignment of any one of the plurality of vertically aligned structures. In some embodiments, the stable film or article substantially maintains its adhesiveness to the biological tissue. In some embodiments, the stable film or article is applicable to a target site of the subject. In some embodiments, the stable film or article maintain its structural and/or mechanical integrity at the target site for at least Ih, at least 2h, at least 3h, at least 5h, including any range between.

[0139] In some embodiments, the stable film or article is rigid under operable conditions. In some embodiments, the stable film is inert to the operable conditions. The operable conditions may be referred to physiological conditions, such as physiological conditions of a mucosal and/or dermal tissue (e.g. oral and/or nasal cavity) such as pH, moisture, enzymatic species, and temperature or any combination thereof).

[0140] In some embodiments, the article of the invention comprising the vertically aligned structures, as described herein is characterized by an enhanced retention at the application site, compared to a control, wherein enhanced is as described herein. In some embodiments, the control is the film having the same composition and being devoid of the vertically aligned structures. [0141] In some embodiments, the article of the invention is for application on top of an injury or damage of a biological tissue (e.g., a wound, an aphtha, a wart, an ulcer). In some embodiments, the biological tissue comprises a mucosal tissue, a dermal tissue, or both.

[0142] In some embodiments, the article is shapeable. In some embodiments, at least one dimension of the article is variable, e.g., by applying stress. In some embodiments, the article is shapeable along at least one dimension, e.g., a length dimension, a width dimension, a radial dimension, a diagonal dimension, and the like. In some embodiments, the article may be shaped and/or elongated, e.g., by a user and/or a medical practitioner, to become elongated, wider, increased in diameter, and/or a combination thereof.

[0143] In some embodiments, the article is foldable. In some embodiments, the article is flexible. In some embodiments, the article is characterized by elasticity. In some embodiments, the article is characterized by elasticity and/or foldability sufficient for application of the article on one or more region of the tissue (e.g. mucous, muscle or dermal tissue).

Uses

[0144] In another aspect, there is provided a method for treating or preventing a condition associated with a tissue damage, the method comprises contacting (or applying) the article of the invention on top of a damaged tissue of a subject, thereby obtaining the article stably bound to the damaged tissue. In some embodiments, stably bound is for a predetermined time period, as described herein. In some embodiments, the method is for sealing or adhering a damaged or injured tissue. In some embodiments, the damaged tissue a damaged mucosal tissue (such as oral and/or nasal cavity). In some embodiments, the method comprises contacting (or applying) the article of the invention on top of a damaged tissue of the subject, thereby adhering the article to the damaged tissue (e.g., to a lesion site). In some embodiment, the article of the innovation for use in the treatment of bleeding, or for wound sealing.

[0145] In some embodiments, the damaged tissue comprises a medical condition associated with a biological tissue damage. In some embodiments, the damaged tissue comprises a medical condition associated with a biological tissue. In some embodiments, the medical condition is a wound. In some embodiments, the damaged tissue comprises an injured tissue (e.g., a physical damage of the mucosal tissue). According to another aspect of some embodiments of the present invention there is provided a method for preventing or treating a medical condition, comprising administering the article of the invention to a subject, thereby preventing, or treating the medical condition.

[0146] In some embodiments, the medical condition is associated with a biological tissue of the subject. In some embodiments, the biological tissue is as described herein. In some embodiments, the biological tissue is a moist tissue. In some embodiments, the biological tissue is a substantially dry tissue (e.g. dermal tissue). In some embodiments, the biological tissue is a mucous, muscle, and/or dermal tissue.

[0147] An active agent to a subject, comprising contacting the article of the invention with a biological tissue of a subject, thereby administering the active agent to the subject. In some embodiments, the article of the invention comprises a pharmaceutically effective amount of the active agent.

[0148] In some embodiments, contacting comprises providing the article and applying the article to a target site of the subject. In some embodiments, applying comprises contacting the adhesive surface with the target site (moist or dry) on or within the biological tissue or organ of the subject. In some embodiments, upon contacting the article with the biological tissue, the vertically aligned structures face or are bound to the biological tissue. In some embodiments, applying comprises pressing the film towards the biological tissue, so as to induce adhesion of the film thereto.

[0149] In some embodiments, the method is for administering a pharmaceutically effective amount of an active agent to the subject (e.g., to a target site on or within the mucosal tissue).

[0150] In some embodiments, the method is for locally administering an active agent. In some embodiments, the method is for topically administering the active agent. In some embodiments. [0151] In some embodiments, the method is for administering the active agent to the target site. In some embodiments, the target site is the application site of the article. In some embodiments, the method is for delivery of the active agent into a mucous or dermal tissue. In some embodiments, the method is for controlled delivery and/or release of the active agent into a mucous or dermal tissue of the subject.

[0152] In some embodiments, the method is for transmucosal and/or transdermal administration of the active agent. In some embodiments, the method is for sustained administration of the active agent. In some embodiments, the method is for sustained release of the active agent to the target site. In some embodiments, the method is for sustained release of the active agent to a biological tissue of the subject. In some embodiments, the biological tissue comprises a mucosal tissue, a dermal tissue, a muscle tissue, and a urinary bladder tissue or any combination thereof.

[0153] According to another aspect of some embodiments of the present invention there is provided a method for preventing or treating a medical condition, comprising administering the article of the invention to a subject, thereby preventing, or treating the medical condition, wherein the article comprises a pharmaceutically effective amount of an active agent. In some embodiments, administering comprising contacting the article with a biological tissue of the subject, as described herein. In some embodiments, the biological tissue comprises a mucosal tissue, a dermal tissue, a muscle tissue, and a urinary bladder tissue or any combination thereof.

[0154] According to an aspect of embodiments of the invention there is provided a medicament comprising one or more articles disclosed herein and a pharmaceutically acceptable carrier.

[0155] According to some embodiments of the invention, the article is packaged in a packaging material and identified in print, in or on the packaging material, for use in the treatment of a medical condition associated with any disease, medical condition, or disorder as described hereinthroughout. [0156] In some embodiments, the disclosed film further comprises a labeling agent. As used herein, the phrase “labeling agent” or “labeling compound” describes a detectable moiety or a probe. The labeling agent may be attached to a portion of the polymer forming the film of the invention, directly or via a spacer. Alternatively, the labeling agent may be encapsulated within the void space within the polymeric film.

[0157] As used herein, the term "subject" refers to any animal (e.g., a mammal), including, but not limited to, humans, non-human primates, rodents, and the like, to which the compositions and methods of the present invention are administered. In some embodiments, the terms "subject" and "patient" are used interchangeably herein in reference to a human subject. In other embodiments, the terms "subject" and "patient" are used interchangeably herein in reference to a non-human subject.

General

[0158] As used herein the term “about” refers to ± 10 %. The terms "comprises", "comprising", "includes", "including", “having” and their conjugates mean "including but not limited to". The term “consisting of means “including and limited to”.

[0159] The term "consisting essentially of" means that the composition, article, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure. The term "consisting essentially of" in conjunction with a chemical composition of a material, article/article portion means that at least 80%, at least 90%, at least 95%, at least 99%, at least 99.9%, between 80 and 99.9%, between 80 and 90%, between 90 and 99.9%, between 90 and 95%, between 80 and 97%, between 80 and 95%, between 80 and 98%, or between 80 and 99.9%, including any range between by weight of the material/article consists of the recited constituents and may optionally include additional ingredients (e.g. excipients, pharmaceutically active agents, etc.) which do not materially alter the basic characteristics of the claimed material/article.

[0160] The word “exemplary” is used herein to mean “serving as an example, instance or illustration”. Any embodiment described as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments and/or to exclude the incorporation of features from other embodiments.

[0161] The word “optionally” is used herein to mean “is provided in some embodiments and not provided in other embodiments”. Any particular embodiment of the invention may include a plurality of “optional” features unless such features conflict.

[0162] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

[0163] As used herein, the term “substantially” refers to at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, including any range or value therebetween.

[0164] Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0165] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging/ranges between” a first indicate number and a second indicate number and “ranging/ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

[0166] As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0167] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

EXAMPLES

[0168] Reference is now made to the following examples which, together with the above descriptions, illustrate the invention in a non-limiting fashion.

Materials and Methods

Materials

[01 9] Following material have been used for the preparation of an exemplary article of the invention and of control articles: crosslinked poly(acrylic acid (Carbomer 941, NF) was purchased from Spectrum (USA), Chitosan (Medium MW), Poly(lactic-co-glycolic acid), with lactide and glycolide at a 50/50, molar ratio (Resomer® RG 503 H), was purchased from Evonik (Germany), acetic acid glacial was purchased from Carlo Erba (Italy), and SYLGARD™ 184 silicone elastomer kits were purchased from Dow Corning (USA). Acetonitrile was purchased from JT Baker (USA), and 2-octyl-CA (Dermabond®), Evicel®, and Surgicel® were purchased from Ethicon Inc. (USA). Duraseal® was purchased from Integra Life-Sciences Corporation (France). Live/dead cell viability assay kits were purchased from Abeam (England), and CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) kits were purchased from Promega (USA). Penicillin-streptomycin, fetal bovine serum, and L-glutamine were purchased from IM Beit HaEmek (Israel). Pre-hydrated collagen membranes were purchased from Vista International Packaging (USA), POLYDINE® was purchased from Dr. Fisher (Israel), and buprenorphine (1.3 mg/mL) was purchased from LogiPharm Ltd. (Israel).

PDMS mold preparation

[0170] A master mold consisting of an array of vertically aligned convergent structures with a total height of 900 pm, was designed using SolidWorks and made with Aluminum and Uddeholm Viking (an oil-air-vacuum-hardening steel). Polydimethylsiloxane (PDMS) solution was poured into the master mold and closed (allowing excess solution to come out of the designated holes) and placed on a standard heated press (Carver Inc.), and pressed at 150°C for 10 minutes. Then, the PDMS mold was separated from the master mold. Figure 1 demonstrates the preparation of the PDMS mold.

Article preparation

[0171 ] Various microneedles (MN) articles were casted using the micro-molding technique. Polymeric solutions were prepared as follows: Carbopol 2%(w/v) in double distilled water (DDW); Pullulan 5% (w/v) in DDW, PLGA 5% (w/v) in acetonitrile; and chitosan 2% (w/v) in 1% (v/v) acetic acid (1% acetic acid in DDW). Then, 500 pL of each of the above-mentioned polymeric solutions were placed in the PDMS mold and pulled into the cavities using a spin coating device (Laurell Technologies, USA; Model WS-650MZ- 23NPPB) at 10,000 rpm for 10 min. The mold was then topped off with polymeric solution (500 pL) followed by a second centrifugation at 10,000 rpm for 10 min. After the centrifugation process, about 5 mL of the polymeric solution was used to fill the mold. Molds were allowed to dry for 48 h at room temperature before MNs were carefully peeled off the mold. The only exception to this procedure was the fabrication of the Carbopol-chitosan MNs where an additional step was carried out: After complete drying of the Carbopol MNs, 5 mL of chitosan solution was poured into the mold (creating the pedestal layer).

[0172] The surface morphology of all polymeric MNs was investigated using a wide-field scanning electron microscope (the Netherlands; Model Quanta 200 FEI ESEM), with an accelerating voltage of 20 kV. Samples were not coated prior to imaging. Figure 2 demonstrates the preparation of the article/control.

PVP-iodine preparation solution

[0173] In a glass cup, put the dried, measured materials (iodine and KI in 1:2 weight ratio) obtaining an iodine solution. Add lOmL of DDW for every gram of KI and mix in a shaker. In a glass cup, mix the dry PVP (M.W. 360kDa) with lOmL of DDW for every gram of PVP obtaining a 10% PVP solution. The two solutions were mixed based on the volumetric ratios presented the table below (table 1). The two solutions were mixed in a shaker at 200[RPM] for at least 4 hours obtaining the PVP-I complex solution.

PVP-iodine micro-needles preparation

[0174] 500pL of a certain PVP-I complex solution was placed in the PDMS master mold and spin coated for 10 minutes at 10,000rpm, this was done additional two times. Then 4 ml of the PVP-I complex solution was added to the PDMS template to cover the template and left to fully dry at room temperature for 2/3 days.

Compression tests and failure morphology

[0175] The vertically aligned structures mechanical behavior was examined to determine their ability to penetrate the skin without breaking. An axial compression load was applied and measured by TAI texture analyzer (Lloyd, AMETEC, USA) equipped with a 50N load cell. The samples were placed on the lower plate using double-sided tape and pressed by the moving upper plate. The pre-test and post-test speeds were 2 mm/min and 1 mm/min respectively, and the trigger force was set at 0. IN. Data were collected until the displacement reached 850 pm, and displayed as force per vertically aligned structure versus extension. After compression, the failure morphology of the compressed MNs patches were characterized by Scanning Electron Microscope (FEIE-SEM Quanta 200, Eindhoven, the Netherlands). Each tested array group contained four samples.

[0176] Given that the polymers are electrically non-conductive, samples were extracted from the obtained film and coated with gold and 5% of palladium to produce a conductive and homogenous 5 nm thin layer on the surface, thus enabling SEM observation. (Exemplary images of the patterned surface before and after the are represented in Figure 1).

Burst pressure

[0177] A standard method for burst strength of surgical sealants of the American Society of Testing and Materials (ASTM F2392-04) was used. In brief, dried collagen membranes (Vista International Packaging, USA) were punctured using a 3 mm puncher and sealed with the tested MN patches. Membranes were then clamped down and pressure was applied by using a Master Dual Pump (Braintree Scientific Inc., USA) to infuse PBS buffer (pH 7.4) into the burst system at a constant rate of 1 mL/min. Pressure in the membrane was monitored throughout the experiment using a pressure gauge (Lutron, USA; Model PS- 9302). Burst pressure was defined as the pressure at which the sealant failed and the membrane started leaking. In all of the burst pressure tests, failure took place around the puncture site, confirming that the internal burst pressure of the collagen membranes was higher than that of the tested materials. The tissue adhesion strength of the MNs was investigated using an EZ50 tensile testing machine (Lloyd Instruments, Denmark) and 1 x 1 cm collagen membrane (Vista International Packaging, USA) immersed in PBS (pH 7.4) for 5 min before the test. Each MN was glued to the stationary crimp using ethyl cyanoacrylate adhesive (3 M, MN, USA). Then, MN were inserted into the wet collagen membranes and a contact force of 150 g was maintained for 4 h, after which the probe was withdrawn from the membrane at a rate of 30 mm/min and the adhesive strength (kPa) was recorded. Adhesive strength was determined as the pressure at which the maximal point, before a sharp decrease, attributed to the disconnection between the tablet and the membrane, was observed. Results are presented as averages ± SD and compared with Surgicel® (n = 5).

Swelling and erosion The swelling and erosion of the different MN patches were measured by incubating each patch in 30 mL of PBS (pH 7.4) at 37 _C with constant shaking (60 rpm). MN patches were weighed at predetermined times after careful blotting with filter paper and returned immediately to the incubator to continue the experiment. The total percent of material weight was calculated by dividing the wet weight of the swollen sample by its initial dry weight prior to incubation (Wd = 40 mg at t = 0). The results are reported as means ± SD (n = 4).

Cell toxicity

[0178] NIH 3T3 fibroblast cells were grown at 37°C in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Gibco-Invitrogen Corp., Grand Island, NY). Cultures were maintained in a 95% air/5% carbon dioxide atmosphere, at 95% relative humidity. MN’s patches were placed inside 24 wells plate and sterilized with a UV lamp for 30 min. The cytotoxicity of the MN’s patches was studied by exposing NIH 3T3 fibroblast cell lines to each material pf the patches. Cytotoxicity was assessed 48h after cells were added, using the MTS kit⁵, and by using live/dead fluorescence viability kit (Abeam, England). The results are reported as means ± SD (n= 4).

Acute in vivo sealing model

[0179] All animal experiments were performed in strict accordance with the Guide for the Care and Use of Laboratory Animals and were approved by the Council for Animal Experiments, Israel Ministry of Health (Animal Ethics Approval No. IL-151-10-2022). Eighteen female Sprague Dawley rats weighing 190-220 g were purchased from Envigo, Israel. All animals were maintained in sterilized cages on a 12-h light/12-h dark cycle with food and water provided. Rats were anesthetized using isoflurane and injected with 0.05 mL of a 1.3 mg/mL long-acting buprenorphine solution.

[0180] Rat abdomens were shaved and sterilized with iodine solution (Polydine, Dr. Fischer, Israel) followed by 70% ethanol. A liver bleeding model was then created44 by exposing the rat liver, placing filter paper on top, and making a 2 mm deep incision at the center of the lobe using a 3 mm puncher. A two-layered MN patch (Carbopol/chitosan) was gently pressed over the bleeding area for 5 seconds. Blood loss was measured by weighing the filter paper around the bleeding area before and at 5-min post-procedure. For comparison, one group of rats was treated with Surgicel® and a third group remained untreated. The results are reported as means ± SD (n = 4).

EXAMPLE 1

MN FABRICATION AND MORPHOLOGY

[0181] Polymeric MNs were fabricated using the micromolding technique. First, a master mold was designed, made of aluminum and Uddeholm Viking using SolidWorks. A poly dimethylsiloxane (PDMS) mold of needles with a total height of 900 pm and base edge of 350 pm was fabricated. Various polymeric solutions were then cast twice in the PDMS mold, spun, and dried for 48 h at room temperature. MNs were configured in four 7 x 7 two- dimensional arrays with an array-to-array spacing of about 1.5 mm (Figure 3c). Scanning electron microscopy (SEM) confirmed a well-structured pyramid-shaped MN configuration with a height and base length of approximately 900 pm and 350 pm, respectively, matching the dimensions of the mold (Figure 3c). Chitosan patches, however, had slightly crooked tips, apparently distorted during removal from the PDMS mold due to the needles' fragile structure.

COMPRESSION TESTS AND FAILURE MORPHOLOGY

[0182] Insertion force, defined as the force required to puncture the top layer of the skin, is an important parameter to estimate the strength of vertically aligned structures (e.g. microneedles (MNs)). The strength of the MNs was determined as the force at which the surface broke and interpreted as the maximum force applied immediately before the force suddenly decrease.

[0183] The polymeric MN patches showed different force-extension curves until the needle broke (Figure 3A) and the strength of the MNs was determined as the force at which the surface break and interpreted as the maximum force applied immediately before the force suddenly decrease (Figure 3B). The results are reported as means ± SD (n = 4). Failure forces for the tested patches ranged between 0.57 N/needle (for pullulan) and 1 N/needle for the PLGA and Carbopol/chitosan MNs (exemplary article of the invention showed the greatest needle strength among the tested samples). SEM analysis was performed to evaluate the effect of compression tests on the morphology and integrity of the needles. Before fracture tests, all needles displayed a complete pyramidal shape integrated into the base substrate. The base and surface of the needles were smooth without the presence of any cracks or fractures, indicating the structural integrity of the patch. After fracture tests, all MNs except for the PLGA showed clear signs of deformation limited to the tips of the needle (Figure 3C). PLGA MNs, on the other hand, broke at the interface between the needles and the base but showed no major deformation or cracks at the tip.

SWELLING AND EROSION

[0184] Swelling and erosion of the mentioned above samples were examined results are demonstrated in Figure 4. While the pullulan MNs dissolved within a few minutes, Carbopol patches swelled by more than 4000% in the first hour and formed a hydrogel-like substance that eroded within the following hour (Figure 4). Chitosan MNs also swelled considerably, around 900% within 2 h, followed by a plateau for 24 h and gradual erosion lasting 22 days.

[0185] The hydrophobic PLGA MNs exhibited minimal swelling, yet lost their initial shape within 7 days and fully degraded within 60 days. Carbopol/chitosan patches showed high swelling degree within 4 h, followed by a plateau for additional 20 h. Then, patches eroded gradually until fully dissolved in 36 days.

[0186] This experiment was also performed on a commercial sealant, fibrin glue, which did not swell and fully degraded in 3 days.

BURST PRESSURE

[0187] To serve as tissue sealants and prevent leakage, MNs must withstand pressure after administration. The burst pressure of thew different MNs were examined. The burst pressure strengths of the single-material MNs were 0 mmHg for PLGA and chitosan, 6 mmHg for Pullulan, and 26 mmHg for pure Carbopol (Figure 3D). Evicel® presented a burst strength of around 22 mmHg and DuraSeal® presented a strength of 100 mmHg (Figure 3D). On the other hand, MNs composed of Carbopol/chitosan exhibited significant higher strength compared with all other MNs tested, with values around 480 mmHg. Since surgical sealants are required to withstand physiological blood pressures ranging from 25 mmHg for capillary bleeding to 120 mmHg for larger blood vessels under normal physiologic conditions, it is clear that only the Carbopol/chitosan MN patches (exemplary article of the invention) possess superior mechanical properties that meet clinical requirements.

[0188] The adhesive strength of the MNs on collagen membrane was further quantified after being stretched at a rate of 30 mm/min. The peak detachment force (kPa) was calculated from the force (N) versus extension (mm) diagram. The pure Carbopol MNs presented the highest adhesive strength, around 90 kPa, followed by the Carbopol/chitosan and the PLGA MNs, both of which had an adhesive strength of 60 kPa, and the Pullulan MNs with 35 kPa. The pure chitosan MNs, as well as the Surgicel® patches exhibited significantly lower strength, around 3 kPa.

CELL TOXICITY

[0189] Viability was tested using the MTS and live and dead assays on fibroblast NIH 3T3 cells. The tested MNs showed minimal toxicity in vitro, which was corroborated with a high percentage of viability. Cells exposed to Chitosan, PLGA and Pullulan patches showed around 100% viability, compared with 50% viability exhibited by the Carbopol patch (both relative to unexposed cells). In all the polymeric patches, live cells were observed around and below the polymeric array. The live/dead assay (Abeam, Cambridge, England) generally mirrored the results of the MTS assay: the majority of cells exposed to Chitosan, PLGA, Pullulan and Carbopol patches were alive. In contrast, almost no living cells were observed in the Dermabond® and Surgicel® groups (Figures 5A-5B).

IN VIVO EFFICACY ASSESSMENT

[0190] MN patches made of chitosan and Carbopol were chosen for this assay since this material combination yielded the most promising results on the burst pressure and swelling and erosion tests. A rat liver bleeding model was used to study the potential of the adhesive MNs as surgical sealants. The model was created by making an incision in a rat liver lobe, after which the lobe began bleeding, and a MN patch was applied directly on the incision area (Figures 6A-6B). The highest blood loss, 132.7 ± 13.8 mg, was observed for the nontreated livers followed by livers treated with Surgicel®, which lost 45.1 ± 12.6 mg, and livers treated with the double-layered MN (Carbopol/chitosan), which lost 13.1 ± 10.4 mg of blood (Figure 6C). Thus, in both treated groups bleeding was significantly reduced. The results also elucidate the superiority of our MN patch over the oxidized cellulose, as bleeding was restricted to the first 5 s, and blood loss was significantly lower than using the commercial hemostat (p < 0.05).

REMARKS

[0191 ] To this end, the inventors prepared and tested varies patches including difffrent polymers. Although the pure Carbopol patches presented the highest adhesive strength (90 kPa) and the second highest average burst pressure values (30 mmHg), pure Carbopol patches swelled excessively and eroded within an hour, making them clinically irrelevant. The patch fabricated from Carbopol and chitosan (i.e. the exemplary article of the invention), that is, Carbopol-based needles and a chitosan-based support layer, overall presented the most promising properties of the various patches. MNs made of this combination exhibited the highest needle failure point (1 N/needle) without compromising elasticity required to maintain the integrity of the needles during penetration.

[0192] The Carbopol/chitosan MNs also had the highest burst strength, 10-fold higher than that of the pure Carbopol MN. The excellent performance of the double-layered MNs can thus be attributed to the role of each layer: the stiff Carbopol needles allow easy and comfortable insertion into wet tissue (fixing the patch via physical and electrostatic interlocking) while the elastic chitosan layer absorbs the interfacial water and becomes a soft hydrogel. The elasticity of chitosan patches and their ability to quickly absorbed water was evident in this study. Thus, a strong reinforcing base layer is crucial for appropriate sealing, as was evident from the burst pressure and the erosion studies. This was also evident by the pure chitosan patch that barely adhered to the collagen membrane and, therefore, failed to withstand any fluid pressure. The fact that these patches degraded within 3 weeks under physiological conditions demonstrated the notion that only by combining the two polymers (i.e. a tissue adhesive polymer such as chitosan and the crosslinked polymer such as Carbopol), a long-lasting adhesive MN patch can be obtained.

Claims

CLAIMS What is claimed is:

1. An article comprising a support and a plurality of vertically aligned structures bound to said support, wherein: said article is composed essentially of biocompatible constituents and comprises (i) a crosslinked polymer, and (ii) a tissue-adhesive polymer; said article is characterized by a retention on or within a biological tissue; the plurality of vertically aligned structures is characterized by (i) an average force at failure of at least 0.5 N per each structure, and (ii) swellability of at least 100%; and an average length of the plurality of vertically aligned structures is between 0.1 and 2 mm; and wherein said support and said plurality of vertically aligned structures are composed of different polymers.

2. The article of claim 1, wherein said support consists essentially of said tissue adhesive polymer.

3. The article of claim 1, wherein each of said plurality of vertically aligned structures is in a form of convergent structure; and wherein said plurality of vertically aligned structures are arranged in an array on top of the support.

4. The article of any one of claims 1 to 3, wherein said plurality of vertically aligned structures consists essentially of said crosslinked polymer.

5. The article of claim 1 or 4, wherein a density of said plurality of vertically aligned structures within said array is between 20 and 1000 units/cm².

6. The article of any one of claims 3 to 5, wherein a center-to-center distance between a pair of adjacent vertically aligned structures within said array is between 10 and 2000 pm.

7. The article of any one of claims 1 to 6, wherein said average force at failure is at least 0.2N force/needle.

8. The article of any one of claims 1 to 7, wherein said array is characterized by an average force at failure of between 4 and 200N.

9. The article of any one of claims 1 to 8, wherein said article is characterized by sealing capability of at least 120 mmHg.

10. The article of claims 1 to 9, wherein each of the plurality of vertically aligned structures has a pyramidal shape; and wherein each of the plurality of vertically aligned structures comprises a base portion in contact with the support, and a top portion distal to the base portion.

11. The article of claim 10, wherein an average width dimension of said base portion is between 50 and 1000 pm.

12. The article of claim 10 or 11, wherein an average width dimension of said top portion is between 0.1 and 10 pm.

13. The article of any one of claims 1 to 12, wherein said crosslinked polymer comprises a crosslinked acrylate -based polymer, and wherein said crosslinked polymer is crosslinked via a covalent crosslinker.

14. The article of claim 13, wherein the covalent crosslinker is a hydroxy alkyl comprising at least two hydroxy groups.

15. The article of any one of claims 1 to 14, wherein said tissue-adhesive polymer comprises alginate, chitosan, pectin, hyaluronic acid, derivates of PEG, cellulose, cellulose derivate, Collagen, Fibrinogen, and non-polyacrylate, including any salt, any derivative, any copolymer, or any combination thereof.

16. The article of any one of claims 1 to 15, wherein said tissue adhesive polymer is chitosan, and wherein said crosslinked acrylate -based polymer is a crosslinked polyacrylate.

17. The article of any one of claims 1 to 16, further comprising a pharmaceutically active ingredient.

18. A method for preventing or treating a medical condition in a subject in need thereof, comprising administering the article of any one of claims 1 to 17 to a subject, thereby preventing or treating said medical condition.

19. The method of claim 18, wherein said medical condition comprises a damage of a biological tissue of said subject.

20. The method of claim 19, wherein said administering comprises contacting the article with the biological tissue of said subject.

21. The method of claim 19 or 20, wherein said biological tissue comprises a mucosal tissue, a dermal tissue, a muscle tissue, or any combination thereof; and wherein said biological tissue is a damaged tissue.