CN112358628B - A method for demolding hydrogels and biomaterials using sacrificial materials - Google Patents

Abstract

The invention discloses a method for demoulding by using a sacrificial material as hydrogel and a biological material, which is based on the reaction of polyethylene glycol diacrylate (PEGDA), Dithiothreitol (DTT) and borax to generate the sacrificial hydrogel which can be dissolved in a high-sugar environment and has good biocompatibility, wherein before the mould is used, the mould is firstly soaked in a mixed solution of 0.28mol/LPEGDA and DTT for 5s, and then is taken out and then is soaked in a 0.1mol/L borax solution for 5s, so that a layer of gel film is quickly formed on the surface of the mould, and then pouring the biological material to be molded, soaking the biological material into a high-sugar culture medium for 10-20 minutes after the material is crosslinked, and enabling the mold to fall off automatically to leave the molded material.

Description

A method for demolding hydrogels and biomaterials using sacrificial materials

technical field

The invention relates to the field of biological materials, in particular to a method for demolding hydrogel and biological materials using sacrificial materials.

Background technique

When using the mold method to prepare hydrogels or other biological materials (need to inoculate cells), the material adheres to the surface of the mold during the stripping process, which is easy to tear, while the existing anti-adhesion agent is easy to adhere to the material and is not easy to wash out , which will affect subsequent biological experiments.

In order to solve this problem, the general solution is to pre-coat a dense anti-stick coating on the mold to facilitate the release of the hydrogel and other biological material contents, but the dense anti-stick coating such as Teflon is coated Layers are toxic to biological materials and can cause material contamination.

To solve this problem, we need a sacrificial hydrogel that is soluble in high sugar environment and has good biocompatibility as a sacrificial sacrificial material to isolate the hydrogel and biomaterials and molds. The gel substrate (Hydrogel) has a three-dimensional network structure, which can quickly swell and not dissolve in the dispersion medium. Due to the existence of the cross-linked network, the gel substrate can swell and retain a large amount of dispersion medium, and the absorption of the dispersion medium is proportional to the coagulation. The degree of crosslinking of the adhesive substrate is closely related. In general, the higher the degree of crosslinking of the gel substrate, the lower the amount of dispersion medium retained. The aggregate state of the gel substrate is neither completely solid nor completely liquid. The behavior of the solid state is that it can maintain a certain shape and volume under certain conditions and has a certain mechanical strength. The behavior of the liquid state is that the solute can be removed from the gel lining. Diffusion or penetration in the bottom. Based on the above-mentioned characteristics of the gel substrate and its unique properties such as flexibility and biocompatibility, it has been widely studied and applied in many fields such as electrical devices, sensors, and biomedicine. However, when the existing gel substrate is used as a sacrificial material in the biomaterial preparation process, it cannot achieve stable coating on the mold due to its amorphous phase state. The processing method in the prior art is to use thick coating. Processing, due to the excessive immersion of the biological material by the thick coating itself, will inevitably interfere with the purity of the biological material in the mold.

SUMMARY OF THE INVENTION

Purpose of the invention: the purpose of the present invention is to improve and innovate for the many defects and deficiencies existing in the above-mentioned background technology or the prior art, and the purpose is to provide a kind of hydrogel and biological material products using sacrificial materials to quickly remove The method of the mould, in order to solve the above-mentioned problems and achieve the above-mentioned purpose of the invention, the present invention is realized through the following design structures and technical solutions:

A method for demolding a hydrogel and a biomaterial using a sacrificial material, comprising the following steps:

S1. Take the mole fraction as 1 part of coupling agent and 1.7 parts of deionized water, and 1 part of coupling agent includes 0.95 part of hydroxymethyl cellulose and 0.05 part of chitosan. Mix the coupling agent into deionized water and stir well to make the coupling agent disperse evenly in the deionized water to obtain the coupling gel auxiliary agent;

S2. Add 0.2 part of guar gum to the coupling gelling aid, heat the mixture to 35°C and stir at a speed of 60r/min for 4min;

S3. Take 12 parts of polyethylene glycol diacrylate and 36 parts of deionized water, mix polyethylene glycol diacrylate into deionized water, heat to 40° C. and stir at 120 r/min for 5 minutes to obtain polyethylene glycol diacrylate. Acrylate aqueous solution, take 16 parts of dithiothreitol and 36 parts of deionized water, mix dithiothreitol into deionized water, and stir at room temperature for 5 minutes at a speed of 300 r/min to obtain an aqueous solution of dithiothreitol;

S4, the polyethylene glycol diacrylate aqueous solution and the dithiothreitol aqueous solution are mixed at room temperature, and the mixed solution obtained in S2 is added at a constant speed while continuously stirring at a rotational speed of 60 r/min;

S5, take 10 parts of borax, 1 part of sodium hydroxide and 89 parts of deionized water, put the borax and sodium hydroxide into the deionized water, and stir at a speed of 600r/min for 2min to obtain a uniform borax solution;

S6. Fully immerse the mold for preparing hydrogels and biomaterials into the mixed solution obtained in S4 and rotate it in the solution at a speed of 12r/min for 5s and then take it out;

S7. Put the mold taken out in S6 into the borax solution, completely immerse it and let it stand for 5s before taking it out;

S8, let stand in the mold taken out in S7 until a gel film is formed on the surface of the mold, and then inject the hydrogel or biological material to be formed, and continue to stand until the cross-linking of the material is completed;

S9, prepare high-glucose medium;

S10. The mold with the cross-linked hydrogel or biomaterial in S8 is completely immersed in the high-sugar medium for 10-20 minutes, and then the hydrogel or biomaterial in the mold is taken out to complete demoulding.

Preferably, the high-glucose medium in S9 includes 4500mg/L glucose, 250mg/L mixed amino acids, 116.6mg/L anhydrous calcium chloride, 0.0013mg/L copper sulfate pentahydrate, 0.05mg/L nine Water ferric nitrate, 0.417mg/L ferrous sulfate heptahydrate, 48.84mg/L anhydrous magnesium sulfate and 6999.5mg/L sodium chloride.

Preferably, the coupling agent in S1 can also be 0.7 part of 2-hydroxyethyl acrylate, 0.03 part of acrylate microgel emulsion, 0.12 part of chitosan and 0.15 part of hydroxyethyl methacrylate.

Compared with the prior art, the present invention has the following beneficial effects: based on the reaction of polyethylene glycol diacrylate (PEGDA), dithiothreitol (DTT) and borax, a kind of bio-phase soluble in high sugar environment is generated. A sacrificial hydrogel with good capacitance is used as a cushion between the biomaterial and the mold. After the biomaterial is prepared, the mold can fall off by itself, leaving a well-shaped material. This method It is convenient and quick, without any modification to the material that needs to be shaped, without adhesion, demoulding in a cell-friendly environment, and can be used in the shaping process of cell-material composites.

Description of drawings

1 is a process flow diagram of a method for demolding a hydrogel and a biomaterial using a sacrificial material according to the present invention.

Detailed ways

The present invention will be further described below in conjunction with the accompanying drawings, but the present invention is not limited to the scope of the described embodiments.

Embodiment 1: a method for demolding a hydrogel and a biomaterial using a sacrificial material, comprising the following steps:

S1. Take the mole fraction of 1 part of coupling agent and 1.7 parts of deionized water, mix the coupling agent into deionized water and stir well to make the coupling agent evenly dispersed in deionized water to obtain coupling gel auxiliary agent, wherein the coupling agent The agent includes 0.7 part of 2-hydroxyethyl acrylate, 0.03 part of acrylate microgel emulsion, 0.12 part of chitosan and 0.15 part of hydroxyethyl methacrylate;

S2. Add 0.2 part of guar gum to the coupling gelling aid, heat the mixture to 35°C and stir at a speed of 60r/min for 4min;

S3. Take 12 parts of polyethylene glycol diacrylate and 36 parts of deionized water, mix polyethylene glycol diacrylate into deionized water, heat to 40° C. and stir at 120 r/min for 5 minutes to obtain polyethylene glycol diacrylate. Acrylate aqueous solution, take 16 parts of dithiothreitol and 36 parts of deionized water, mix dithiothreitol into deionized water, and stir at room temperature for 5 minutes at a speed of 300 r/min to obtain an aqueous solution of dithiothreitol;

S4, the polyethylene glycol diacrylate aqueous solution and the dithiothreitol aqueous solution are mixed at room temperature, and the mixed solution obtained in S2 is added at a constant speed while continuously stirring at a rotational speed of 60 r/min;

S5, take 10 parts of borax, 1 part of sodium hydroxide and 89 parts of deionized water, put the borax and sodium hydroxide into the deionized water, and stir at a speed of 600r/min for 2min to obtain a uniform borax solution;

S6. Fully immerse the mold for preparing hydrogels and biomaterials into the mixed solution obtained in S4 and rotate it in the solution at a speed of 12r/min for 5s and then take it out;

S7. Put the mold taken out in S6 into the borax solution, completely immerse it and let it stand for 5s before taking it out;

S8, let stand in the mold taken out in S7 until a gel film is formed on the surface of the mold, and then inject the hydrogel or biological material to be formed, and continue to stand until the cross-linking of the material is completed;

S9. Prepare a high-glucose medium, which includes 4500mg/L glucose, 250mg/L mixed amino acids, 116.6mg/L anhydrous calcium chloride, 0.0013mg/L copper sulfate pentahydrate, 0.05mg/L L of ferric nitrate nonahydrate, 0.417mg/L of ferrous sulfate heptahydrate, 48.84mg/L of anhydrous magnesium sulfate and 6999.5mg/L of sodium chloride;

S10. The mold with the cross-linked hydrogel or biomaterial in S8 is completely immersed in the high-sugar medium for 10-20 minutes, and then the hydrogel or biomaterial in the mold is taken out to complete demoulding.

Embodiment 2: a method for demolding a hydrogel and a biomaterial using a sacrificial material, comprising the following steps:

S1. Take the mole fraction of 1 part of coupling agent and 1.7 parts of deionized water, mix the coupling agent into deionized water and stir well to make the coupling agent evenly dispersed in deionized water to obtain coupling gel auxiliary agent, wherein the coupling agent The agent includes 0.95 part of hydroxymethyl cellulose and 0.05 part of chitosan;

S2. Add 0.2 part of guar gum to the coupling gelling aid, heat the mixture to 35°C and stir at a speed of 60r/min for 4min;

S3. Take 12 parts of polyethylene glycol diacrylate and 36 parts of deionized water, mix polyethylene glycol diacrylate into deionized water, heat to 40° C. and stir at 120 r/min for 5 minutes to obtain polyethylene glycol diacrylate. Acrylate aqueous solution, take 16 parts of dithiothreitol and 36 parts of deionized water, mix dithiothreitol into deionized water, and stir at room temperature for 5 minutes at a speed of 300 r/min to obtain an aqueous solution of dithiothreitol;

S4, the polyethylene glycol diacrylate aqueous solution and the dithiothreitol aqueous solution are mixed at room temperature, and the mixed solution obtained in S2 is added at a constant speed while continuously stirring at a rotational speed of 60 r/min;

S5, take 10 parts of borax, 1 part of sodium hydroxide and 89 parts of deionized water, put the borax and sodium hydroxide into the deionized water, and stir at a speed of 600r/min for 2min to obtain a uniform borax solution;

S6. Fully immerse the mold for preparing hydrogels and biomaterials into the mixed solution obtained in S4 and rotate it in the solution at a speed of 12r/min for 5s and then take it out;

S7. Put the mold taken out in S6 into the borax solution, completely immerse it and let it stand for 5s before taking it out;

S8, let stand in the mold taken out in S7 until a gel film is formed on the surface of the mold, and then inject the hydrogel or biological material to be formed, and continue to stand until the cross-linking of the material is completed;

S9. Prepare a high-glucose medium, which includes 4500mg/L glucose, 250mg/L mixed amino acids, 116.6mg/L anhydrous calcium chloride, 0.0013mg/L copper sulfate pentahydrate, 0.05mg/L L of ferric nitrate nonahydrate, 0.417mg/L of ferrous sulfate heptahydrate, 48.84mg/L of anhydrous magnesium sulfate and 6999.5mg/L of sodium chloride;

S10. The mold with the cross-linked hydrogel or biomaterial in S8 is completely immersed in the high-sugar medium for 10-20 minutes, and then the hydrogel or biomaterial in the mold is taken out to complete demoulding.

Embodiment 3: A method for demolding hydrogel and biomaterial using traditional anti-adhesion agent, comprising the following steps:

S1, flatly coating the inner wall of the mold to be used with synthetic silica powder to form an insulating cushion layer of 0.01-0.028 mm;

S2. Coat 0.015mm of hydrogel on the insulating cushion and wait for it to cure;

S3. Inject the hydrogel or biological material to be formed into the mold processed in S2, and continue to stand until the cross-linking of the material is completed.

The demolded hydrogels or biomaterials in Example 1 and Example 2 were analyzed as a whole, and the interface integrity of the contact surface with the mold and the contamination degree of impurities at a depth of 0.5 mm were tested. The parameters obtained are as follows.

interface integrity Impurity contamination Example 1 99.28% 0.015% Example 2 99.71% 0.072% Example 3 98.33% 0.285%

It can be seen from the above data that 0.7 part of 2-hydroxyethyl acrylate, 0.03 part of acrylate microgel emulsion, 0.12 part of chitosan and 0.15 part of hydroxyethyl methacrylate are used as coupling agents. The prepared sacrificial material The fusion compatibility with biomaterials is better, which can effectively reduce the degree of contamination of the surface interface of biomaterials, but the resulting sacrificial materials have insufficient cross-linking rigidity, and the interface integrity of biomaterials will be damaged to a certain extent when demolding; 0.95 part of methyl cellulose and 0.05 part of chitosan are used as coupling agents, the prepared sacrificial material has better cross-linking rigidity, and the interface integrity is higher when the biomaterial is demolded, but the fusion compatibility with the biomaterial is poor. The degree of contamination on the surface and interface of the material will increase, but compared with the traditional anti-adhesion agent, the hydrogel and biomaterial obtained by using sacrificial materials as hydrogels and biomaterials by demoulding have better interface integrity and better quality. The impurity contamination degree has been significantly optimized, and the optimized ratio is above 300%.

The advantage of this technical solution is that it is based on the reaction of polyethylene glycol diacrylate (PEGDA), dithiothreitol (DTT) and borax to generate a sacrificial product that is soluble in high sugar environments and has good biocompatibility. Hydrogel, using this sacrificial hydrogel as the cushion between the biomaterial and the mold, the mold can fall off by itself after the biomaterial is prepared, leaving the shaped material. The material that needs to be shaped has any modification, does not stick, is demolded in a cell-friendly environment, and can be used in the shaping process of cell-material composites.

The preferred embodiments of the present invention have been described above in detail. It should be understood that those skilled in the art can make many modifications and changes according to the concept of the present invention without creative efforts. Therefore, any technical solutions that can be obtained by those skilled in the art through logical analysis, reasoning or limited experiments on the basis of the prior art according to the concept of the present invention shall fall within the protection scope determined by the claims.

Claims (3)

1. a kind of method that uses sacrificial material to carry out demoulding for hydrogel and biological material, it is characterized in that: comprise the following steps: S1. Take the mole fraction of 1 part of coupling agent and 1.7 parts of deionized water, 1 part of coupling agent includes 0.95 part of hydroxymethyl cellulose and 0.05 part of chitosan, mix the coupling agent into deionized water and stir well to make it The coupling agent is uniformly dispersed in deionized water to obtain coupling gel auxiliary; S2. Add 0.2 part of guar gum to the coupling gelling aid, heat the mixture to 35°C and stir at a speed of 60r/min for 4min; S3. Take 12 parts of polyethylene glycol diacrylate and 36 parts of deionized water, mix polyethylene glycol diacrylate into deionized water, heat to 40° C. and stir at 120 r/min for 5 minutes to obtain polyethylene glycol diacrylate. Acrylate aqueous solution, take 16 parts of dithiothreitol and 36 parts of deionized water, mix dithiothreitol into deionized water, and stir at room temperature for 5 minutes at a speed of 300 r/min to obtain an aqueous solution of dithiothreitol; S4, the polyethylene glycol diacrylate aqueous solution and the dithiothreitol aqueous solution are mixed at room temperature, and the mixed solution obtained in S2 is added at a constant speed while continuously stirring at a rotational speed of 60 r/min; S5, take 10 parts of borax, 1 part of sodium hydroxide and 89 parts of deionized water, put the borax and sodium hydroxide into the deionized water, and stir at a speed of 600r/min for 2min to obtain a uniform borax solution; S6. Fully immerse the mold for preparing hydrogels and biomaterials into the mixed solution obtained in S4 and rotate it in the solution at a speed of 12r/min for 5s and then take it out; S7. Put the mold taken out in S6 into the borax solution, completely immerse it and let it stand for 5s before taking it out; S8, let stand in the mold taken out in S7 until a gel film is formed on the surface of the mold, and then inject the hydrogel or biological material to be formed, and continue to stand until the cross-linking of the material is completed; S9, prepare high-glucose medium; S10. The mold with the cross-linked hydrogel or biomaterial in S8 is completely immersed in the high-sugar medium for 10-20 minutes, and then the hydrogel or biomaterial in the mold is taken out to complete demoulding. 2. a kind of method that uses sacrificial material according to claim 1 to be hydrogel and biological material to carry out demoulding, it is characterized in that: high glucose medium in S9 comprises the glucose of 4500mg/L, the mixed amino acid of 250mg/L, 116.6mg/L of calcium chloride anhydrous, 0.0013mg/L of copper sulfate pentahydrate, 0.05mg/L of ferric nitrate nonahydrate, 0.417mg/L of ferrous sulfate heptahydrate, 48.84mg/L of anhydrous sulfuric acid Magnesium and 6999.5mg/L of sodium chloride. 3. a kind of method that uses sacrificial material to be hydrogel and biological material to carry out demoulding according to claim 1, it is characterized in that: coupling agent in S1 can also be 0.7 part of acrylic acid-2-hydroxyethyl ester, Acrylate microgel emulsion 0.03 part, chitosan 0.12 part and hydroxyethyl methacrylate 0.15 part.

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