阅读说明：本技术 一种用于面部皮肤支架的技术方法 (Technical method for facial skin support ) 是由 王心营 欧阳则军 于 2019-10-25 设计创作，主要内容包括：本发明公开了一种用于面部皮肤支架的技术方法,第一步,制备生物打印墨水；第二步,使用计算机辅助软件SolidWorks设计面部皮肤支架数据模型；第三步,低温条件下进行3D生物打印,形成面部皮肤支架初型；第四步,将第三步得到的面部皮肤支架初型进行冷冻干燥成型；本发明的用于面部皮肤支架的技术方法,结合低温3D打印与冷冻干燥技术制备面部皮肤支架,可在保持支架材料本身性能的基础上,还可以实现支架材料结构的定制。(The invention discloses a technical method for a facial skin bracket, which comprises the following steps of preparing biological printing ink; secondly, designing a facial skin bracket data model by using computer aided software SolidWorks; thirdly, performing 3D biological printing under a low-temperature condition to form a facial skin support prototype; fourthly, freeze-drying and forming the primary shape of the facial skin support obtained in the third step; the technical method for the facial skin stent of the invention combines the low-temperature 3D printing and freeze drying technology to prepare the facial skin stent, and can realize the customization of the stent material structure on the basis of keeping the performance of the stent material.)

1. A technical method for facial skin scaffolding, comprising the steps of:

firstly, preparing biological printing ink;

secondly, designing a facial skin bracket data model by using computer aided software SolidWorks;

thirdly, performing 3D biological printing under a low-temperature condition to form a facial skin support prototype;

and fourthly, freeze-drying and molding the facial skin support prototype obtained in the third step.

2. A technical method for facial skin scaffolds according to claim 1, wherein the bioprinting ink consists of silk fibroin and carboxymethyl chitosan, and the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5.

3. The technical method for facial skin scaffolds according to claim 1, wherein the bio-printing ink is composed of bioactive glass, silk fibroin and carboxymethyl chitosan, wherein the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5, and the mass ratio of bioactive glass to the mixture of silk fibroin and chitosan is 1: 10.

4. The technical method for facial skin scaffold as claimed in claim 2, wherein the silk fibroin solution is prepared by the following method,

step one, Na with the mass fraction of 0.05 percent is used₂CO₃Boiling the water solution for 4 ~ 6 times, each time for 0.8 ~ 1 hr, at 100 ~ 110 deg.C and bath ratio of 1: 100;

step two, fully washing with distilled water, and drying at 45 ~ 55 ℃ to obtain pure silk cellulose;

step three, using a mixture of a molar ratio of 1: 2: 8 CaCl₂、CH₃CH₂OH and H₂Dissolving the O ternary solvent, wherein the mass ratio of the solution to the raw silk is 10: 1, and stirring and dissolving the solution for 1 ~ 2 hours at 42 ℃ of ~ 65 ℃ to obtain a mixed solution;

step four, injecting the cooled solution into a cellulose dialysis membrane, intercepting the solution with the molecular weight of 1 ten thousand ~ 1.2 ten thousand, and dialyzing the solution in running water for 3 days;

and step five, air-drying and concentrating the dialyzed solution to obtain a silk fibroin concentrated aqueous solution, and storing at 4 ℃ for later use.

5. A technical process for facial skin scaffolds, according to claim 3, characterized in that the bioactive glass is prepared as follows:

step one, adding ethyl orthosilicate, triethyl phosphate and calcium nitrate tetrahydrate into a mixed solution of 2mo1/L hydrochloric acid solution and a certain volume of deionized water in sequence every 30min, and uniformly stirring by magnetic force to obtain a uniform and transparent bioactive glass sol solution;

step two, preparing wet gel, namely standing and aging the bioactive glass sol solution prepared in the step one at room temperature, and fully hydrolyzing and polycondensing to form wet gel;

step three, preparing dry gel, drying the wet gel in a constant-temperature drying oven at the temperature of 110 ~ 128 ℃ for 3 days, and evaporating the solvent to obtain the dry gel;

step four, preparing sol-gel bioactive glass particles, and carrying out heat treatment on the xerogel by using a box-type electric furnace at 670 ℃ to obtain granular sol-gel bioactive glass;

step five, preparing bioactive glass particle powder, and performing wet ball milling on bioactive glass particles by using alcohol as a dispersing agent; drying, grading on a vibrating screen, and taking bioactive glass particles with the particle size of less than 88 mu m for later use.

6. The technical method for facial skin scaffolds as claimed in claim 2, wherein the preparation method of the biological printing ink consisting of silk fibroin and carboxymethyl chitosan comprises mixing the purified carboxymethyl chitosan with silk fibroin solution using purified water, stirring uniformly, and vacuum defoaming to obtain chitosan/silk fibroin mixed solution, wherein the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5.

7. A technical method for a facial skin scaffold as claimed in claim 3, wherein the preparation method of the bio-printing ink consisting of bioactive glass, silk fibroin and carboxymethyl chitosan comprises mixing the purified carboxymethyl chitosan and silk fibroin solution with purified water, stirring uniformly, and vacuum-defoaming to obtain chitosan/silk fibroin mixed solution, wherein the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5, mixing the bioactive glass particles and the chitosan/silk fibroin mixed solution according to the mass ratio of bioactive glass to chitosan/silk fibroin mixed solution being 1:10, stirring uniformly, and vacuum-defoaming to obtain bioactive glass/chitosan/silk fibroin mixed solution.

8. The technical method for forming the facial skin scaffold as claimed in claim 1, wherein the freeze-drying molding in the fourth step comprises the following specific operation steps of firstly, standing the facial skin scaffold in a refrigerator at-20 ℃ overnight, then placing the facial skin scaffold in a freeze dryer, carrying out vacuum freeze drying at-80 ℃ ~ -60 ℃ for 24 hours to obtain a three-dimensional porous composite scaffold, then, soaking the obtained composite scaffold material in an aqueous methanol solution with the volume fraction of 90% for 30 minutes to obtain a water-insoluble three-dimensional porous composite scaffold, and then, drying in an oven at 37 ℃ for 4 hours to obtain the water-insoluble bioactive glass/chitosan/silk fibroin composite scaffold.

Technical Field

The invention relates to a technical method for a facial skin bracket, and belongs to the technical field of medical cosmetology.

Background

Currently, in the field of artificial tissue engineering skin scaffolds, materials for skin scaffold preparation are roughly classified into two types: one is artificially synthesized biological high molecular material, mainly comprising polyglycolide, polycaprolactone, polyhydroxyalkanoate, polycarbonate and other polyester materials; the other is natural biological derived material, such as collagen, chitosan, hyaluronic acid, carboxymethyl chitosan, silk fibroin, etc. Most skin scaffolds are fabricated using biomaterials by electrospinning techniques or cross-linking; although the nanofiber obtained by electrostatic spinning provides a proper surface morphology for the adhesion and growth of cells, which is beneficial to the adhesion and growth of cells on the scaffold, the skin scaffold prepared by electrostatic spinning is not beneficial to the migration and proliferation of cells in the depth direction due to small aperture; the porous skin scaffold prepared by crosslinking has uncontrollable pore size and porosity and poor mechanical strength, and can also hinder cell growth, which limits the application of the artificial skin scaffold in the field of medical treatment of skin injury. The principle of the freeze-drying method is thermally induced phase separation, which means that polymer solution, emulsion or hydrogel formed by a high polymer and a dispersion system is pre-frozen and shaped at a certain temperature, a continuous interpenetration structure is formed in the freezing process due to the growth of ice crystal grains, and when the ice crystals are dried and sublimated in a vacuum environment, holes left by sublimation of the ice crystals are maintained, so that a sponge-type porous scaffold is formed; the scaffold prepared by the method can retain the performance of raw materials to the maximum extent, the specific surface area of the pores is large, the operation is simple, but the pore distribution in the scaffold is difficult to control, the size of the pore diameter is small, and the mechanical stability is low. The invention combines the freeze drying method with the 3D printing technology, and can accurately regulate and control the aperture, the porosity, the connectivity and the specific surface area of the bracket.

Disclosure of Invention

In order to solve the problems, the invention provides a technical method for a facial skin stent, the facial skin stent is prepared by combining low-temperature 3D printing and freeze drying technologies, and the customization of the structure of the stent material can be realized on the basis of keeping the performance of the stent material.

The technical method for the facial skin support comprises the following steps:

firstly, preparing biological printing ink;

secondly, designing a facial skin bracket data model by using computer aided software SolidWorks;

thirdly, performing 3D biological printing under a low-temperature condition to form a facial skin support prototype;

and fourthly, freeze-drying and molding the facial skin support prototype obtained in the third step.

As a preferred embodiment, the bioprinting ink is composed of silk fibroin and carboxymethyl chitosan, and the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5.

As a preferred embodiment, the bio-printing ink is composed of bioactive glass, silk fibroin and carboxymethyl chitosan, wherein the mass ratio of the chitosan to the silk fibroin is 1: 9 ~ 5: 5, and the mass ratio of the bioactive glass to the mixture of the silk fibroin and chitosan is 1: 10.

Further, the preparation method of the silk fibroin solution is as follows,

step one, Na with the mass fraction of 0.05 percent is used₂CO₃Boiling the water solution for 4 ~ 6 times, each time for 0.8 ~ 1 hr, at 100 ~ 110 deg.C and bath ratio of 1: 100;

step two, fully washing with distilled water, and drying at 45 ~ 55 ℃ to obtain pure silk cellulose;

step three, using a mixture of a molar ratio of 1: 2: 8 CaCl₂、CH₃CH₂OH and H₂Dissolving the O ternary solvent, wherein the mass ratio of the solution to the raw silk is 10: 1, and stirring and dissolving the solution for 1 ~ 2 hours at 42 ℃ of ~ 65 ℃ to obtain a mixed solution;

step four, injecting the cooled solution into a cellulose dialysis membrane, intercepting the solution with the molecular weight of 1 ten thousand ~ 1.2 ten thousand, and dialyzing the solution in running water for 3 days;

and step five, air-drying and concentrating the dialyzed solution to obtain a silk fibroin concentrated aqueous solution, and storing at 4 ℃ for later use.

Further, the preparation method of the bioactive glass comprises the following steps:

step one, adding ethyl orthosilicate, triethyl phosphate and calcium nitrate tetrahydrate into a mixed solution of 2mo1/L hydrochloric acid solution and a certain volume of deionized water in sequence every 30min, and uniformly stirring by magnetic force to obtain a uniform and transparent bioactive glass sol solution;

step two, preparing wet gel, namely standing and aging the bioactive glass sol solution prepared in the step one at room temperature, and fully hydrolyzing and polycondensing to form wet gel;

step three, preparing dry gel, drying the wet gel in a constant-temperature drying oven at the temperature of 110 ~ 128 ℃ for 3 days, and evaporating the solvent to obtain the dry gel;

step four, preparing sol-gel bioactive glass particles, and carrying out heat treatment on the xerogel by using a box-type electric furnace at 670 ℃ to obtain granular sol-gel bioactive glass;

step five, preparing bioactive glass particle powder, and performing wet ball milling on bioactive glass particles by using alcohol as a dispersing agent; drying, grading on a vibrating screen, and taking bioactive glass particles with the particle size of less than 88 mu m for later use.

Further, the preparation method of the biological printing ink consisting of silk fibroin and carboxymethyl chitosan comprises the following steps of mixing purified carboxymethyl chitosan and silk fibroin solution by using purified water, uniformly stirring, and defoaming in vacuum to obtain a chitosan/silk fibroin mixed solution, wherein the mass ratio of chitosan to silk fibroin is 1: 9 ~ 5: 5.

The preparation method of the biological printing ink composed of the bioactive glass, the silk fibroin and the carboxymethyl chitosan comprises the following steps of firstly mixing the purified carboxymethyl chitosan and the silk fibroin solution by using purified water, uniformly stirring, and carrying out vacuum defoaming to obtain a chitosan/silk fibroin mixed solution, wherein the mass ratio of the chitosan to the silk fibroin is 1: 9 ~ 5: 5, then mixing the bioactive glass particles and the chitosan/silk fibroin mixed solution according to the mass ratio of the bioactive glass to the chitosan/silk fibroin mixed solution being 1:10, uniformly stirring, and carrying out vacuum defoaming to obtain the bioactive glass/chitosan/silk fibroin mixed solution.

Further, the freeze-drying molding in the fourth step comprises the specific operation steps of firstly, standing the facial skin scaffold prototype in a refrigerator at the temperature of 20 ℃ below zero overnight, then placing the facial skin scaffold prototype in a freeze dryer, carrying out vacuum freeze drying for 24 hours at the temperature of 80 ℃ below zero ~ ℃ to 60 ℃ to obtain a three-dimensional porous composite scaffold, then, soaking the obtained composite scaffold material in an aqueous methanol solution with the volume fraction of 90% for 30 minutes to obtain a water-insoluble three-dimensional porous composite scaffold, and then, drying the water-insoluble three-dimensional porous composite scaffold in an oven at the temperature of 37 ℃ for 4 hours to obtain the water-insoluble bioactive glass/chitosan/silk fibroin composite scaffold.

Compared with the prior art, the technical method for preparing the facial skin scaffold combines the low-temperature 3D printing technology and the freeze drying technology to prepare the facial skin scaffold, is simple and convenient to operate, and can accurately regulate and control the pore diameter, porosity, connectivity and specific surface area of the scaffold; the requirements of the laser curing molding technology on the photosensitive material can be avoided, the influence of the high-temperature environment of the high-temperature melting process on the biological activity of the material can be avoided, the problem that the biological activity of the raw material is kept in 3D printing is solved, and the personalized customized manufacturing of the bracket can be realized.

Detailed Description