CN104147645A - Injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction and preparation method thereof - Google Patents
Injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction and preparation method thereof Download PDFInfo
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- CN104147645A CN104147645A CN201410210634.XA CN201410210634A CN104147645A CN 104147645 A CN104147645 A CN 104147645A CN 201410210634 A CN201410210634 A CN 201410210634A CN 104147645 A CN104147645 A CN 104147645A
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Abstract
The invention provides an injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction and a preparation method thereof. The preparation method of the polypeptide injectable porous microcarrier material is characterized in that the polypeptide injectable porous microcarrier is prepared by an emulsion method under a condition of no pore-foaming agent or adding the pore-foaming agent, wherein the concentration of the polypeptide is 0.1-1.0 percent by weight, the pore diameter is 20-60mu m, and the porosity is 80-90 percent. The invention firstly provides the porous injectable microcarrier constructed by synthesizing polypeptide materials. The polypeptide injectable microcarrier is applicable in the field of tissue engineering, a relatively good environment can be provided to cell adhesion and proliferation, and in-vivo tissue regeneration and reconstruction can be realized.
Description
Technical field
The present invention relates to a kind of injectable porous microcarrier material and preparation method thereof, particularly a kind of injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction and preparation method thereof.
Background technology
First Microcarrier Culture Techniques is proposed in 1967 by Van Wezel, is to generally acknowledge in the world at present to carry out the tool of the mass cell cultivation cell culture technology of development prospect.In recent years, microcarrier also receives much attention in field of tissue engineering technology.Microcarrier has a series of advantages as injectable type support for organizational project, comprises and maintains cell differentiation phenotype, exempts surgical procedures, Reduction surgery wound and alleviate patient's misery etc.The material of preparing microcarrier has natural material and synthetic material.Natural material has sodium alginate, agarose, gelatin, hyaluronic acid, chitosan and collagen etc.Chen etc. with emulsion method prepared biocompatibility chitosan microcarrier as biomaterial for organizational project cell carrier, the shortcomings such as but natural material has, and source is batch unstable, extraction, purge process may be subject to polluting, spread disease, bad mechanical property, physicochemical property are uncontrollable, have limited its application in organizational project.Conventional synthetic material has polylactide and its copolymer, polycaprolactone etc.Newman etc. prepare finishing LN and load simultaneously the polylactic acid microcarrier of RA, have studied embryo cells P19 in microcarrier superficial growth, but this type of material degradation product is generally acid, easily cause in body local temperature chaotic and cause inflammatory reaction.
In view of the above problems, the present invention adopts similar human body protein in structure, and on composition, the poly-polypeptide material of similar extracellular matrix is as the novel biomaterial of preparing microcarrier.Poly-polypeptide material is the polymer that aminoacid is formed by connecting by peptide chain, and it can be degraded into single amino acid by protease in vivo, and catabolite is needed by human material, safety non-toxic, and can be absorbed by body.Poly-polypeptide material is owing to having the features such as good biological activity, biocompatibility, biodegradability, high-hydrophilic, low antigenicity or immunogenicity, having become a class new bio medical material with broad prospect of application.Have no report about this class microcarrier and in the research of field of tissue engineering technology application at present, the present invention attempts the porous injectable microcarrier of preparation containing poly-polypeptide, and regenerate for in-vivo tissue, experimental result shows, cell adhesion, multiplication capacity can be significantly improved containing poly-polypeptide microcarrier, and in-vivo tissue regeneration can be promoted.
Summary of the invention
One of object of the present invention is to provide a kind of injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction.
Another object of the present invention is to provide the preparation method of this microcarrier material.
The preparation method of the poly-polypeptide injectable porous microcarrier material that the present invention proposes, is under the condition without porogen or interpolation porogen, prepares the porous injectable microcarrier of poly-polypeptide by emulsion method.Wherein poly-peptide concentration is 0.1-1.0wt%, and aperture is 20-60 μ m, and porosity is 80-90%.
In the present invention, poly-polypeptide is synthetic polyamino acid Ester.
The preparation method of the poly-polypeptide injectable porous microcarrier material proposing in the present invention, concrete steps are as follows:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 0.1-1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
opre-cooling;
2. poly-polypeptide being dissolved in to compound concentration in dichloromethane is the solution of 0.1-1 wt%, is stirred to poly-polypeptide and dissolves completely under room temperature, obtains colourless transparent solution;
3. step 1 gained 100 mL pre-cooling PVA solution are placed in to ice-water bath, under stirring, poly-step 2 gained 20 mL polypeptide polymer solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
The present invention obtains the injectable microcarrier for in-vivo tissue regeneration and reconstruction by above-mentioned preparation method, and this microcarrier material comprises polyamino acid methyl ester, polyamino acid ethyl ester and polyamino acid benzyl ester material; Described poly-polypeptide material passes through cold-emulsion legal system for the compound microcarrier of porous; The compound microcarrier of described porous can be used as cell microcarrier for in-vivo tissue regeneration and rebuilds.
Major advantage of the present invention is: propose first to synthesize poly-polypeptide class material construction porous injectable microcarrier.Microcarrier has loose structure, and a large amount of cells of energy load, have good cell compatibility and histocompatibility.To gather polypeptide injectable microcarrier for field of tissue engineering technology, and can provide better environment for cell adhesion and propagation, and can realize in-vivo tissue regeneration and rebuild.
Brief description of the drawings
Fig. 1 is the electron scanning micrograph of poly-polypeptide injectable porous microcarrier.Microcarrier has good spherical structure and loose and porous structure as shown in the figure, and hole connects continuously, and the scope of aperture and porosity is also at 20-60 μ m and 80-90%.
Fig. 2 is the fluorescence co-focusing figure growing on poly-polypeptide injectable porous microcarrier.Green fluorescence point shows chondrocyte condition of production on microcarrier.Left figure is the fluorescence co-focusing figure of Growth of Cells 1 d, and cell is attached on microcarrier surface at first; Right figure is the fluorescence co-focusing figure of Growth of Cells 7 d, as shown in the figure cells fill microcarrier surface and inner.Along with inoculated and cultured time lengthening, green fluorescence is counted out and is increased, and shows that chondrocyte is along with incubation time extends and breeds.
Fig. 3 is the digital photo of poly-polypeptide injectable porous microcarrier for regenerating bone or cartilage.As seen from the figure, through mouse bare subcutaneous injection culturing in vivo after 8 weeks, formed there is translucent white, smooth surface and elastic type cartilaginous tissue, tissue surface is not found bulk band vascular tissue and undesired tissue.
Detailed description of the invention
In order further to understand the present invention, below in conjunction with embodiment, injectable porous microcarrier material for in-vivo tissue regeneration and reconstruction provided by the invention and preparation method thereof is described, protection scope of the present invention is not limited by the following examples.
embodiment 1:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid methyl ester being dissolved in to compound concentration in dichloromethane is the solution of 1 wt%, is stirred to polyamino acid methyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid methyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 2:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid methyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.5 wt%, is stirred to polyamino acid methyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid methyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 3:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid methyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.1 wt%, is stirred to polyamino acid methyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid methyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 4:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid ethyl ester being dissolved in to compound concentration in dichloromethane is the solution of 1 wt%, is stirred to polyamino acid ethyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid ethyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 5:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid ethyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.5 wt%, is stirred to polyamino acid ethyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid ethyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 6:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid ethyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.1 wt%, is stirred to polyamino acid ethyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100 mL pre-cooling PVA solution are placed in to ice-water bath, under stirring, 20mL polyamino acid ethyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 7:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid benzyl ester being dissolved in to compound concentration in dichloromethane is the solution of 1 wt%, is stirred to polyamino acid benzyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid benzyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 8:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid benzyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.5 wt%, is stirred to polyamino acid benzyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid benzyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 9:
1. polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 1 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling.
2. polyamino acid benzyl ester being dissolved in to compound concentration in dichloromethane is the solution of 0.1 wt%, is stirred to polyamino acid benzyl ester and dissolves completely under room temperature, obtains colourless transparent solution; Step 1 gained 100mL pre-cooling PVA solution is placed in to ice-water bath, under stirring, 20mL polyamino acid benzyl ester solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
embodiment 10:in 100 mL culture dishs, 5 × 10
6individual chondrocyte mixes with 1 mL microsphere density, and adds 20% FBS containing 10 mL DMEM.Then, in incubator, with 75 rpm rotating speed jolting culture dish 6 h, then add 20 mL media transfer to 37
oc, 5% CO
2, long-term cultivation in 100% saturated humidity incubator, the next day of later, change culture fluid.
embodiment 11:porous microcarrier is mixed with chondrocyte suspension, and be expelled to 7 d nude mice by subcutaneous in age.No. 23 needle injection 2 times for both sides, every nude mice back contain respectively 2 × 10 at every turn
6the cell microcarrier complex of individual cell (0.5 mL) and 1 mg microcarrier, injects 18 mices altogether, after eight weeks, cartilaginous tissue newly-generated nude mice by subcutaneous is used respectively to Toluidine blue staining, and immunohistochemical staining is for biochemical analysis.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (3)
1. a preparation method for poly-polypeptide injectable porous microcarrier material, is characterized in that, under the condition without porogen or interpolation porogen, preparing the porous injectable microcarrier of poly-polypeptide by emulsion method; Wherein poly-peptide concentration is 0.1-1.0wt%, and aperture is 20-60 μ m, and porosity is 80-90%.
2. the poly-polypeptide injectable porous microcarrier material of stating according to claim 1, is characterized in that described poly-polypeptide is synthetic polyamino acid Ester, as polyglutamic acid methyl ester, ethyl ester, benzyl ester etc.; Polylysine methyl ester, ethyl ester, benzyl ester etc.; Poly arginine methyl ester, ethyl ester, benzyl ester etc. and other amino acid esters of birdsing of the same feather flock together.
3. prepare the preparation method of porous microcarrier according to the poly-polypeptide polymer of stating according to claim 1, it is characterized in that concrete steps are as follows:
(1). polyvinyl alcohol (PVA) is dissolved in to deionized water and is mixed with the solution that concentration is 0.1-1.0 wt%, 90-95
oafter being stirred to PVA under C and dissolving completely, naturally cool to room temperature, 4
oc pre-cooling;
(2). it is the solution of 0.1-1.0 wt% that poly-polypeptide is dissolved in to compound concentration in dichloromethane, is stirred to poly-polypeptide and dissolves completely under room temperature, obtains colourless transparent solution;
(3). step 1 gained 100 mL pre-cooling PVA solution are placed in to ice-water bath, under stirring, poly-step 2 gained 20mL polypeptide polymer solution is joined in PVA solution fast, mechanical agitation is volatilized completely down to dichloromethane, filters, deionized water wash is removed after PVA, obtains porous microcarrier.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826161A (en) * | 2015-04-28 | 2015-08-12 | 上海大学 | Poly(amino acid) based porous microgel material for tissue engineering and preparation method thereof |
| CN106479967A (en) * | 2016-12-01 | 2017-03-08 | 中国科学院长春应用化学研究所 | A kind of cell microcarrier and preparation method thereof |
| CN108659605A (en) * | 2018-05-31 | 2018-10-16 | 句容市东山纸品厂 | A kind of preparation method and applications of ink anti-pollution modifying agent |
| CN110639058A (en) * | 2019-10-30 | 2020-01-03 | 上海大学 | Acicular HA/PBLG porous composite microcarrier material for bone tissue engineering and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250390A (en) * | 2011-05-25 | 2011-11-23 | 天津大学 | Alginate hydrogel microcarrier and preparation method thereof |
| CN102558601A (en) * | 2012-01-04 | 2012-07-11 | 上海理工大学 | Preparation method of high-strength porous chitosan microcarrier |
| US20130156820A1 (en) * | 2011-12-16 | 2013-06-20 | Gc Corporation | Production method of tissue regeneration material and tissue regeneration material |
-
2014
- 2014-05-19 CN CN201410210634.XA patent/CN104147645A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250390A (en) * | 2011-05-25 | 2011-11-23 | 天津大学 | Alginate hydrogel microcarrier and preparation method thereof |
| US20130156820A1 (en) * | 2011-12-16 | 2013-06-20 | Gc Corporation | Production method of tissue regeneration material and tissue regeneration material |
| CN102558601A (en) * | 2012-01-04 | 2012-07-11 | 上海理工大学 | Preparation method of high-strength porous chitosan microcarrier |
Non-Patent Citations (4)
| Title |
|---|
| 周燕等: "微载体培养技术的研究与进展", 《中国组织工程研究与临床康复》 * |
| 张凤菊等: "聚合物载体及其在细胞培养方面的应用", 《北京生物医学工程》 * |
| 方建军: "基于PLGA和PBLG的可注射微载体制备及其组织工程应用", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 方建军: "基于PLGA和PBLG的可注射微载体制备及其组织工程应用", 《中国博士学位论文全文数据库 医药卫生科技辑》, 15 May 2014 (2014-05-15) * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826161A (en) * | 2015-04-28 | 2015-08-12 | 上海大学 | Poly(amino acid) based porous microgel material for tissue engineering and preparation method thereof |
| CN106479967A (en) * | 2016-12-01 | 2017-03-08 | 中国科学院长春应用化学研究所 | A kind of cell microcarrier and preparation method thereof |
| CN108659605A (en) * | 2018-05-31 | 2018-10-16 | 句容市东山纸品厂 | A kind of preparation method and applications of ink anti-pollution modifying agent |
| CN110639058A (en) * | 2019-10-30 | 2020-01-03 | 上海大学 | Acicular HA/PBLG porous composite microcarrier material for bone tissue engineering and preparation method thereof |
| CN110639058B (en) * | 2019-10-30 | 2022-02-08 | 上海大学 | Acicular HA/PBLG porous composite microcarrier material for bone tissue engineering and preparation method thereof |
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Application publication date: 20141119 |