CN103976805B - The manufacture method of hydrogel/high molecular polymer thin film muscular tissue support - Google Patents
The manufacture method of hydrogel/high molecular polymer thin film muscular tissue support Download PDFInfo
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- CN103976805B CN103976805B CN201410235459.XA CN201410235459A CN103976805B CN 103976805 B CN103976805 B CN 103976805B CN 201410235459 A CN201410235459 A CN 201410235459A CN 103976805 B CN103976805 B CN 103976805B
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Abstract
The invention discloses a kind of hydrogel/high molecular polymer thin film muscular tissue support manufacturing method, obtain three dimensional scaffold structure by the mode be layering.First blood vessel network structure and directed runner two kinds of moulds are designed and manufactured according to the architectural feature of muscle, polymer directional thin film is produced at use electrostatic spinning technique, to first prepare support single layer planar structure, then described scaffold three-dimensional structure is being added to by stacked for other each layer shelf layer, acquisition is wrapped up, hydrogel/high molecular polymer film frame again with the high molecular polymer thin membrance mirror support that outermost layer adheres to.Hydrogel manufactures blood vessel network and directed runner, the oriented growth of cell can be realized by directed runner, blood vessel network pipeline can realize the supply of nutrition and oxygen, the thin polymer film of peripheral parcel has directive construction, also the oriented growth of peripheral cell can be realized, the thin film extended, realizes the fixing of support by binding, stitching etc.
Description
Technical field
The present invention relates to a kind of preparation method of muscular tissue support, be specifically related to the manufacture method of a kind of hydrogel/high molecular polymer thin film muscular tissue support.
Background technology
Muscle comprises cardiac muscle, smooth muscle, skeletal muscle, and the identical point of three has sarcostyle, blood vessel and nerve, and myocyte becomes to align.Difference is distribution, the form of tissue in three, the difference to some extent such as sarcostyle quantity form, function.Skeletal muscle is soft tissue relatively large in human body, and account for 40% ~ 45% of body weight, it is connected with bone photo by tendon, when receiving stimulation, by contraction and the diastole of skeletal muscle, realizes the function of body movement.Normal muscle regeneration is mainly completed by muscle satellite cell (satellitecell, SC), but SC quantity is few, and increases with the age and decline.For serious or that area is larger muscle injury, SC is not enough to the object reaching reparation, and wound surface is finally replaced by fibrous tissue and forms cicatrix, and cause wound difficulty more, body loses motor function.At present for large-area muscle injury, treatment means the most general is clinically musculocutaneous flap grafting, the method is to larger for district's damage, and limited source, muscular tissue support provides possibility for overcoming these shortcomings, but the preparation process of existing muscular tissue support is more difficult, the muscular tissue scaffold three-dimensional with blood vessel structure builds difficulty, and muscular tissue support is fixing inconvenient.
Summary of the invention
The object of the invention is to the shortcoming overcoming above-mentioned prior art, provide the manufacture method of a kind of hydrogel/high molecular polymer thin film muscular tissue support, the muscular tissue scaffold three-dimensional that the method is convenient to blood vessel structure builds and fixes.
For achieving the above object, the manufacture method of hydrogel of the present invention/high molecular polymer thin film muscular tissue support comprises the following steps:
1) the inner tubular structure model of hydrogel/high molecular polymer thin film muscular tissue support is drawn, then machine-processed for N number of resin piece by photocureable rapid shaping according to described inner tubular structure model;
2) silica gel solution is prepared, then by step 1) N number of resin piece of obtaining is separately fixed on Silicon moulds, then the silica gel solution that will prepare injects Silicon moulds, and then remove the bubble in silica gel solution by evacuation, leave standstill curing and demolding, obtain N shell silica gel mould;
3) prepare high molecular polymer PLGA solution, then electrostatic spinning is carried out to the high molecular polymer PLGA solution prepared, obtain thin polymer film;
4) to step 2) hydrophilic treated must be carried out by N shell silica gel mould;
5) prepare gelatin solution, then by the gelatin solution for preparing by evacuation de-soak;
6) glutamine transaminage solution is prepared, then glutamine transaminage solution is divided into N part, simultaneously by step 5) gelatin solution after de-soak that obtains is divided into N part, by first part of glutamine transaminage solution and first part of gelatin solution mix homogeneously after de-soak, obtain first part of mixed solution A, then first part of mixed solution A is injected in ground floor silica gel mould, and by step 3) thin polymer film that obtains is bonded in the bottom surface of ground floor silica gel mould, the gel phase that thin polymer film and first part of mixed solution A are formed bonds, and then through solidification and the demoulding, obtain support single layer planar structure,
7) by second part of glutamine transaminage solution and second part of gelatin solution mix homogeneously after de-soak, obtain second part of mixed solution A, then second part of mixed solution A is injected in second layer silica gel mould, and the bottom of the gel formed in the top of support single layer planar structure and second part of mixed solution A is bonding, and then carry out the demoulding;
8) the gelatin solution mix homogeneously after de-soak by X part glutamine transaminage solution and X part, obtain X part mixed solution A, then X part mixed solution A is injected in X layer silica gel mould, and the bottom of the gel formed in the top of the gel of X-1 part mixed solution A formation and X part mixed solution A is bonding, and then carry out the demoulding, wherein, 3≤X≤N;
9) step 8 is repeated), obtain 3-D solid structure, then by step 3) thin polymer film that obtains is wrapping to the surface of 3-D solid structure, after insulation, obtains hydrogel/high molecular polymer thin film muscular tissue support.
Step 2) in preparation silica gel solution concrete steps be: silica gel and the firming agent of getting certain volume, be then added in silica gel by firming agent, obtain silica gel solution, and wherein, the volume ratio of silica gel and firming agent is 100: 1.5-2;
Step 2) in resin piece be fixed on Silicon moulds by double faced adhesive tape, place 24-28h solidification.
Step 3) in solvent in PLGA macromolecule polymer solution be deionized water, the mass concentration of PLGA macromolecule polymer solution is 15 ~ 20%;
Step 3) in carry out voltage in the process of electrostatic spinning to the high molecular polymer PLGA solution prepared be 10-15kV, flow velocity is 0.1 ~ 0.3ml/min, and roller rotating speed is 2800 ~ 3600r/min.
Step 4) in the concrete operations that described N shell silica gel mould carries out hydrophilic treated be: described N shell silica gel mould is placed into plasma cleaning 2 ~ 5min in plasma cleaner.
Step 5) in preparation gelatin solution detailed process be: take a certain amount of gelatin, gelatin is poured in the phosphate buffer of certain volume, be melt 30-40min under the condition of 60-70 DEG C in temperature, stir, obtain gelatin solution, in gelatin solution, the mass concentration of gelatin is 10% ~ 20%;
The mass concentration of mixed solution A GLN transaminase is 1 ~ 1.2%;
Step 6) in solidification hardening time be 4 ~ 6min;
Step 6) in step 3) to pass in and out direction consistent for the orientation direction of thin polymer film and the blood vessel that obtain.
The time that each part mixed solution A forms gel is 4 ~ 6min.
Step 9) in insulation concrete operations be: 3-D solid structure is placed in 37 DEG C of calorstats be incubated 4 ~ 5h.
The present invention has following beneficial effect:
The manufacture method of hydrogel of the present invention/high molecular polymer thin film muscular tissue support is in preparation process, first prepare N shell silica gel mould, again described N shell silica gel mould is obtained 3-D solid structure by the method be layering, then thin polymer film is wrapped on described 3-D solid structure, thus obtain hydrogel/high molecular polymer thin film muscular tissue support, preparation process is simple, is easy to operation.The three-dimensional that the present invention adopts the method be layering to realize hydrogel/high molecular polymer film frame builds, hydrogel/high molecular polymer thin film muscular tissue the support of preparation has blood vessel network structure and directive construction, blood vessel network can be cells with nutrient in hydrogel/high molecular polymer film frame and oxygen, directive construction can realize the oriented growth of cell, thin polymer film is wrapped up described 3-D solid structure, thin polymer film self has directive construction on the one hand, for cell directional growth provides topological structure, thin polymer film periphery parcel can increase the intensity of hydrogel on the other hand, hydrogel/high molecular polymer film frame fixed function can be realized simultaneously, thus the three-dimensional blood vessel network solving muscular tissue support builds, the problem that cell directional growth and support are fixed.
Accompanying drawing explanation
Fig. 1 is the structural representation of hydrogel/high molecular polymer thin film muscular tissue support prepared by the present invention.
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is described in further detail:
With reference to figure 1, the manufacture method of hydrogel of the present invention/high molecular polymer thin film muscular tissue support comprises the following steps:
1) the inner tubular structure model of hydrogel/high molecular polymer thin film muscular tissue support is drawn, then machine-processed for N number of resin piece by photocureable rapid shaping according to described inner tubular structure model;
2) silica gel solution is prepared, then by step 1) N number of resin piece of obtaining is separately fixed on Silicon moulds, then the silica gel solution that will prepare injects Silicon moulds, and then remove the bubble in silica gel solution by evacuation, leave standstill curing and demolding, obtain N shell silica gel mould;
3) prepare high molecular polymer PLGA solution, then electrostatic spinning is carried out to the high molecular polymer PLGA solution prepared, obtain thin polymer film;
4) to step 2) hydrophilic treated must be carried out by N shell silica gel mould;
5) prepare gelatin solution, then by the gelatin solution for preparing by evacuation de-soak;
6) glutamine transaminage solution is prepared, then glutamine transaminage solution is divided into N part, simultaneously by step 5) gelatin solution after de-soak that obtains is divided into N part, by first part of glutamine transaminage solution and first part of gelatin solution mix homogeneously after de-soak, obtain first part of mixed solution A, then first part of mixed solution A is injected in ground floor silica gel mould, and by step 3) thin polymer film that obtains is bonded in the bottom surface of ground floor silica gel mould, the gel phase that thin polymer film and first part of mixed solution A are formed bonds, and then through solidification and the demoulding, obtain support single layer planar structure,
7) by second part of glutamine transaminage solution and second part of gelatin solution mix homogeneously after de-soak, obtain second part of mixed solution A, then second part of mixed solution A is injected in second layer silica gel mould, and the bottom of the gel formed in the top of support single layer planar structure and second part of mixed solution A is bonding, and then carry out the demoulding;
8) the gelatin solution mix homogeneously after de-soak by X part glutamine transaminage solution and X part, obtain X part mixed solution A, then X part mixed solution A is injected in X layer silica gel mould, and the bottom of the gel formed in the top of the gel of X-1 part mixed solution A formation and X part mixed solution A is bonding, and then carry out the demoulding, wherein, 3≤X≤N;
9) step 8 is repeated), obtain 3-D solid structure, then by step 3) thin polymer film that obtains is wrapping to the surface of 3-D solid structure, after insulation, obtains hydrogel/high molecular polymer thin film muscular tissue support.
Step 2) in preparation silica gel solution concrete steps be: silica gel and the firming agent of getting certain volume, be then added in silica gel by firming agent, obtain silica gel solution, and wherein, the volume ratio of silica gel and firming agent is 100: 1.5-2;
Step 2) in resin piece be fixed on Silicon moulds by double faced adhesive tape, place 24-28h solidification.
Step 3) in solvent in PLGA macromolecule polymer solution be deionized water, the mass concentration of PLGA macromolecule polymer solution is 15 ~ 20%;
Step 3) in carry out voltage in the process of electrostatic spinning to the high molecular polymer PLGA solution prepared be 10-15kV, flow velocity is 0.1 ~ 0.3ml/min, and roller rotating speed is 2800 ~ 3600r/min.
Step 4) in the concrete operations that described N shell silica gel mould carries out hydrophilic treated be: described N shell silica gel mould is placed into plasma cleaning 2 ~ 5min in plasma cleaner.
Step 5) in preparation gelatin solution detailed process be: take a certain amount of gelatin, gelatin is poured in the phosphate buffer of certain volume, be melt 30-40min under the condition of 60-70 DEG C in temperature, stir, obtain gelatin solution, in gelatin solution, the mass concentration of gelatin is 10% ~ 20%;
The mass concentration of mixed solution A GLN transaminase is 1 ~ 1.2%;
Step 6) in solidification hardening time be 4 ~ 6min;
Step 6) in step 3) to pass in and out direction consistent for the orientation direction of thin polymer film and the blood vessel that obtain.
The time that each part mixed solution A forms gel is 4 ~ 6min.
Step 9) in insulation concrete operations be: 3-D solid structure is placed in 37 DEG C of calorstats be incubated 4 ~ 5h.
Claims (7)
1. a manufacture method for hydrogel/high molecular polymer thin film muscular tissue support, is characterized in that, comprise the following steps:
1) the inner tubular structure model of hydrogel/high molecular polymer thin film muscular tissue support is drawn, then machine-processed for N number of resin piece by photocureable rapid shaping according to described inner tubular structure model;
2) silica gel solution is prepared, then by step 1) N number of resin piece of obtaining is separately fixed on Silicon moulds, then the silica gel solution that will prepare injects Silicon moulds, and then remove the bubble in silica gel solution by evacuation, leave standstill curing and demolding, obtain N shell silica gel mould;
3) prepare high molecular polymer PLGA solution, then electrostatic spinning is carried out to the high molecular polymer PLGA solution prepared, obtain thin polymer film;
4) to step 2) hydrophilic treated must be carried out by N shell silica gel mould;
5) prepare gelatin solution, then by the gelatin solution for preparing by evacuation de-soak;
6) glutamine transaminage solution is prepared, then glutamine transaminage solution is divided into N part, simultaneously by step 5) gelatin solution after de-soak that obtains is divided into N part, by first part of glutamine transaminage solution and first part of gelatin solution mix homogeneously after de-soak, obtain first part of mixed solution A, then first part of mixed solution A is injected in ground floor silica gel mould, and by step 3) thin polymer film that obtains is bonded in the bottom surface of ground floor silica gel mould, the gel phase that thin polymer film and first part of mixed solution A are formed bonds, and then through solidification and the demoulding, obtain support single layer planar structure,
7) by second part of glutamine transaminage solution and second part of gelatin solution mix homogeneously after de-soak, obtain second part of mixed solution A, then second part of mixed solution A is injected in second layer silica gel mould, and the bottom of the gel formed in the top of support single layer planar structure and second part of mixed solution A is bonding, and then carry out the demoulding;
8) the gelatin solution mix homogeneously after de-soak by X part glutamine transaminage solution and X part, obtain X part mixed solution A, then X part mixed solution A is injected in X layer silica gel mould, and the bottom of the gel formed in the top of the gel of X-1 part mixed solution A formation and X part mixed solution A is bonding, and then carry out the demoulding, wherein, 3≤X≤N;
9) step 8 is repeated), obtain 3-D solid structure, then by step 3) thin polymer film that obtains is wrapping to the surface of 3-D solid structure, after insulation, obtains hydrogel/high molecular polymer thin film muscular tissue support.
2. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, is characterized in that,
Step 2) in preparation silica gel solution concrete steps be: silica gel and the firming agent of getting certain volume, be then added in silica gel by firming agent, obtain silica gel solution, and wherein, the volume ratio of silica gel and firming agent is 100:1.5-2;
Step 2) in resin piece be fixed on Silicon moulds by double faced adhesive tape, place 24-28h solidification.
3. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, it is characterized in that, step 3) in solvent in PLGA macromolecule polymer solution be deionized water, the mass concentration of PLGA macromolecule polymer solution is 15 ~ 20%;
Step 3) in carry out voltage in the process of electrostatic spinning to the high molecular polymer PLGA solution prepared be 10-15kV, flow velocity is 0.1 ~ 0.3ml/min, and roller rotating speed is 2800 ~ 3600r/min.
4. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, is characterized in that,
Step 4) in the concrete operations that described N shell silica gel mould carries out hydrophilic treated be: described N shell silica gel mould is placed into plasma cleaning 2 ~ 5min in plasma cleaner.
5. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, it is characterized in that, step 5) in preparation gelatin solution detailed process be: take a certain amount of gelatin, gelatin is poured in the phosphate buffer of certain volume, be melt 30-40min under the condition of 60-70 DEG C in temperature, stir, obtain gelatin solution, in gelatin solution, the mass concentration of gelatin is 10% ~ 20%.
6. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, is characterized in that, the time that each part mixed solution A forms gel is 4 ~ 6min.
7. the manufacture method of hydrogel/high molecular polymer thin film muscular tissue support according to claim 1, is characterized in that, step 9) in the concrete operations of insulation be: 3-D solid structure is placed in 37 DEG C of calorstats and is incubated 4 ~ 5h.
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| CN104959988B (en) * | 2015-05-29 | 2017-05-10 | 浙江大学 | Muscle driving module based on temperature responsiveness hydrogel |
| CN107050521B (en) * | 2017-04-27 | 2020-10-09 | 天新福（北京）医疗器材股份有限公司 | Double-layer collagen dermal scaffold and preparation method thereof |
| CN107296669B (en) * | 2017-05-18 | 2019-11-26 | 西安交通大学 | A kind of outer hanging bracket of degradable tracheae and its indirect 3D printing method |
| CN107151646B (en) * | 2017-05-18 | 2021-07-09 | 西安交通大学 | Active biological battery construction method based on generating cells |
| CN110344151B (en) * | 2019-07-25 | 2024-04-16 | 东华大学 | Bionic scaffold simulating natural tendon tissue fiber hierarchical structure and preparation method thereof |
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