CN1314460C - Method for compounding aquo-gel to porous tissue engineering rack - Google Patents
Method for compounding aquo-gel to porous tissue engineering rack Download PDFInfo
- Publication number
- CN1314460C CN1314460C CNB2004100529844A CN200410052984A CN1314460C CN 1314460 C CN1314460 C CN 1314460C CN B2004100529844 A CNB2004100529844 A CN B2004100529844A CN 200410052984 A CN200410052984 A CN 200410052984A CN 1314460 C CN1314460 C CN 1314460C
- Authority
- CN
- China
- Prior art keywords
- hydrogel
- porous
- support
- colloidal sol
- porous support
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 33
- 238000013329 compounding Methods 0.000 title abstract 2
- 239000000017 hydrogel Substances 0.000 claims abstract description 86
- 230000010261 cell growth Effects 0.000 claims abstract description 18
- 239000003102 growth factor Substances 0.000 claims abstract description 13
- 229920000747 poly(lactic acid) Polymers 0.000 claims description 59
- 239000004626 polylactic acid Substances 0.000 claims description 59
- 229920001817 Agar Polymers 0.000 claims description 48
- 239000008272 agar Substances 0.000 claims description 46
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 41
- 239000000661 sodium alginate Substances 0.000 claims description 41
- 235000010413 sodium alginate Nutrition 0.000 claims description 41
- 229940005550 sodium alginate Drugs 0.000 claims description 41
- 150000001875 compounds Chemical class 0.000 claims description 34
- 210000004027 cell Anatomy 0.000 claims description 32
- 210000001612 chondrocyte Anatomy 0.000 claims description 25
- 210000001519 tissue Anatomy 0.000 claims description 25
- 108010010803 Gelatin Proteins 0.000 claims description 18
- 229920000159 gelatin Polymers 0.000 claims description 18
- 239000008273 gelatin Substances 0.000 claims description 18
- 235000019322 gelatine Nutrition 0.000 claims description 18
- 235000011852 gelatine desserts Nutrition 0.000 claims description 18
- 239000008363 phosphate buffer Substances 0.000 claims description 14
- 102000008186 Collagen Human genes 0.000 claims description 13
- 108010035532 Collagen Proteins 0.000 claims description 13
- 229920001436 collagen Polymers 0.000 claims description 13
- 230000005484 gravity Effects 0.000 claims description 13
- 102000018233 Fibroblast Growth Factor Human genes 0.000 claims description 12
- 108050007372 Fibroblast Growth Factor Proteins 0.000 claims description 12
- 229940126864 fibroblast growth factor Drugs 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 claims description 7
- 239000001527 calcium lactate Substances 0.000 claims description 7
- 229960002401 calcium lactate Drugs 0.000 claims description 7
- 235000011086 calcium lactate Nutrition 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 210000000963 osteoblast Anatomy 0.000 claims description 7
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 229920001661 Chitosan Polymers 0.000 claims description 5
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 claims description 4
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 claims description 4
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 229940112869 bone morphogenetic protein Drugs 0.000 claims description 4
- 239000001506 calcium phosphate Substances 0.000 claims description 4
- 229920001577 copolymer Polymers 0.000 claims description 4
- 210000002950 fibroblast Anatomy 0.000 claims description 4
- 210000003494 hepatocyte Anatomy 0.000 claims description 4
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims description 4
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims description 4
- 229920001610 polycaprolactone Polymers 0.000 claims description 4
- 239000004632 polycaprolactone Substances 0.000 claims description 4
- 229920000642 polymer Polymers 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 229920002635 polyurethane Polymers 0.000 claims description 4
- 239000004814 polyurethane Substances 0.000 claims description 4
- 210000000329 smooth muscle myocyte Anatomy 0.000 claims description 4
- 230000001131 transforming effect Effects 0.000 claims description 4
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims description 4
- 229940078499 tricalcium phosphate Drugs 0.000 claims description 4
- 229910000391 tricalcium phosphate Inorganic materials 0.000 claims description 4
- 235000019731 tricalcium phosphate Nutrition 0.000 claims description 4
- -1 phosphoglyceride disodium salt Chemical class 0.000 claims description 2
- 229920002503 polyoxyethylene-polyoxypropylene Polymers 0.000 claims description 2
- 239000011833 salt mixture Substances 0.000 claims description 2
- 239000000499 gel Substances 0.000 abstract description 6
- 238000002360 preparation method Methods 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000024245 cell differentiation Effects 0.000 abstract 1
- 239000008279 sol Substances 0.000 description 47
- 230000001476 alcoholic effect Effects 0.000 description 24
- 230000008676 import Effects 0.000 description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 16
- 230000000694 effects Effects 0.000 description 12
- PMMYEEVYMWASQN-DMTCNVIQSA-N Hydroxyproline Chemical compound O[C@H]1CN[C@H](C(O)=O)C1 PMMYEEVYMWASQN-DMTCNVIQSA-N 0.000 description 10
- 239000011148 porous material Substances 0.000 description 10
- PMMYEEVYMWASQN-UHFFFAOYSA-N dl-hydroxyproline Natural products OC1C[NH2+]C(C([O-])=O)C1 PMMYEEVYMWASQN-UHFFFAOYSA-N 0.000 description 9
- 229960002591 hydroxyproline Drugs 0.000 description 9
- FGMPLJWBKKVCDB-UHFFFAOYSA-N trans-L-hydroxy-proline Natural products ON1CCCC1C(O)=O FGMPLJWBKKVCDB-UHFFFAOYSA-N 0.000 description 9
- 230000008859 change Effects 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 239000012153 distilled water Substances 0.000 description 7
- 238000012545 processing Methods 0.000 description 7
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 6
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 6
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 6
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 6
- 210000002744 extracellular matrix Anatomy 0.000 description 6
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 5
- 229910001424 calcium ion Inorganic materials 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 4
- 229940020947 fluorescein sodium Drugs 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 239000002131 composite material Substances 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 230000017423 tissue regeneration Effects 0.000 description 3
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 description 2
- 102100031706 Fibroblast growth factor 1 Human genes 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 229920002994 synthetic fiber Polymers 0.000 description 2
- QGZCUOLOTMJILH-UHFFFAOYSA-N 2h-tetrazol-2-ium;bromide Chemical compound [Br-].C1=N[NH+]=NN1 QGZCUOLOTMJILH-UHFFFAOYSA-N 0.000 description 1
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 1
- 229920001287 Chondroitin sulfate Polymers 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 229940059329 chondroitin sulfate Drugs 0.000 description 1
- 230000030944 contact inhibition Effects 0.000 description 1
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 150000002327 glycerophospholipids Chemical class 0.000 description 1
- 210000003709 heart valve Anatomy 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000003361 porogen Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000002145 thermally induced phase separation Methods 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
Images
Landscapes
- Materials For Medical Uses (AREA)
Abstract
The present invention discloses a method for compounding hydrogel to a porous tissue engineering rack, which comprises the following steps: under the state of sol, hydrogel is led into a porous rack, and then an external condition is altered to convert the sol into gel to obtain the porous rack with compounded hydrogel. As required, cell growth factors for cell growth and differentiation can be added into the hydrogel sol. The rack prepared by the method has the excellent mechanical performance of the porous rack and the excellent biocompatible performance of a hydrogel rack simultaneously. The present invention has a wide application range and is suitable for various structures, various porous racks composed of various structures, hydrogel and various cells. The present invention has the advantages of simple preparation process and good repeatability. The prepared rack can be widely applied to the field of tissue engineering.
Description
Technical field
The present invention relates to hydrogel is compound to method in the porous tissue engineering scaffold.
Background technology
Utilize natural or synthetic material analog cell epimatrix, it is combined with cell, so promotion and inducing cell growth formative tissue or organ, repair or the damaged purpose of reconstruction human organ to reach, become the method for widely admitting in the Tissue Engineering Study.Cytoskeletal preparation and combine with intercellular is the major issue that organizational project need solve.
According to the morphosis of tissue regeneration support, a large amount of at present supports that use are divided into two classes: porous support and hydrogel support.Porous support is made up of the gap of material and storeroom, and under the dry state situation, this gap is generally air; In cell culture environment, this gap is generally culture medium.At present, developed the porous support that several different methods prepares various structures, as porogen method, thermally induced phase separation, fibrage method, emulsion lyophilization, microsphere method of piling, 3 dimension-print processes etc.Porous support has the degradation rate of mechanical strength and easy-regulating preferably; Easy macroscopical plastotype also can keep good macroshape during tissue regeneration.But also there is significant disadvantages in this class support.Under condition of in vitro culture, most somatic cells have the adherent growth characteristic, but the growth of being surrounded by substrate under the adherent growth of cell and its interior state has tangible difference, easier the dedifferenting of cell and lose biological function.Cell owing to the restriction that is subjected to the growing space comes in contact inhibition, can't continue division after long-time growth division on the hole wall, cause the cell number that can adhere on the porous support little than the tissue of native state, and the inequality that distributes.In addition, because the biologically inert of most synthetic materials also must carry out the improvement of biocompatibility to support, increased preparation technology's complexity.The hydrogel support also is to be made of material and gap, and this material is generally natural or synthetic hydrophilic macromolecule.Polymeric material forms very thin gel network, contains a large amount of water therebetween.But these water are not freely, are in highly bonded state because of being subjected to the high molecular constraint of aquation on every side.Therefore, hydrogel class support and extracellular matrix have similar structure and performance, and especially the natural macromolecular hydrogel material has good biocompatibility, can more approach intravital growing environment for the growth of cell and differentiation provide.Simultaneously, cell is dispersed in the hydrogel, has bigger growing space.There is significant disadvantages equally in this class support, and promptly mechanical strength is generally relatively poor, and macroscopical plastotype difficulty also can't keep good macroshape during tissue regeneration, therefore can not effectively control the shape of regeneration organ; Simultaneously, operability is relatively poor relatively.
Summary of the invention
The purpose of this invention is to provide a kind of operating process method in the porous tissue engineering scaffold that hydrogel is compound to simple, applied widely, to obtain to have simultaneously the tissue engineering bracket of good mechanical properties and cell compatibility.
The method that hydrogel is compound in the porous tissue engineering scaffold of the present invention is under collosol state hydrogel to be imported in the porous support, changes external condition then and makes collosol and gelization, obtains the porous support of composite aquogel.
Of the present invention hydrogel is compound to method in the porous tissue engineering scaffold, three kinds of technical solutions are arranged.
Scheme one, hydrogel is compound to method in the porous tissue engineering scaffold, specifically may further comprise the steps:
1) under 4 ℃～120 ℃ agar is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the colloidal sol bulking value concentration that obtains is 0.5～5%;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the hydrogel colloidal sol, hydrogel is adsorbed onto porous support inside;
4) support that will be compounded with colloidal sol was placed 10 minutes down at 15 ℃～40 ℃ at least, the porous support of hydrogel that promptly obtained load.
Usually in order to improve the biology performance of this compound rest, can in agar, sneak into gelatin or/and cell growth factor, the ratio of gelatin consumption and agar consumption is 1: 1～1: 100, the ratio of cell growth factor consumption and agar consumption is 1: 10,000～1: 1,000,000.Said cell growth factor can be that alkalescence facilitates fibroblast growth factor, acidity to facilitate fibroblast growth factor, bone morphogenetic protein(BMP) or transforming factor β.
Scheme two, hydrogel is compound to method in the porous tissue engineering scaffold, specifically may further comprise the steps:
1) under 4 ℃～120 ℃ sodium alginate is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the colloidal sol bulking value concentration that obtains is 0.5～5%;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the hydrogel colloidal sol, hydrogel is adsorbed onto porous support inside;
4) will be compounded be placed in 0.1%～2% calcium lactate solution at least 5 minutes of colloidal sol, the porous support of hydrogel that promptly obtained load.
Usually in order to improve the biology performance of this compound rest, can in sodium alginate, sneak into gelatin or/and cell growth factor, the ratio of gelatin consumption and sodium alginate consumption is 1: 1～1: 100, the ratio of cell growth factor consumption and sodium alginate consumption is 1: 10,000～1: 1,000,000.Said cell growth factor can be that alkalescence facilitates fibroblast growth factor, acidity to facilitate fibroblast growth factor, bone morphogenetic protein(BMP) or transforming factor β.
Scheme three, hydrogel is compound to method in the porous tissue engineering scaffold, specifically may further comprise the steps:
1) under 4 ℃～120 ℃ with the phosphoglyceride disodium salt mixture of chitosan and 1～5 times of weight, or polyoxyethylene-polyoxypropylene copolymer, or PNIPAM is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the bulking value concentration that obtains colloidal sol is 0.5～5% again;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the hydrogel colloidal sol, hydrogel is adsorbed onto porous support inside;
4) will be compounded be placed under 30 ℃～40 ℃ conditions at least 5 minutes of colloidal sol, the porous support of hydrogel that promptly obtained load.
Said porous support can be the polymer class porous support in above-mentioned three kinds of technical solutions of the present invention, comprise polylactic acid, collagen, lactic acid-ethanol copolymer, polycaprolactone, polyurethane or polystyrene holder, can be the ceramic-like porous support also, comprise porous hydroxyapatite, tricalcium phosphate or bio-vitric support.
Said cell includes but not limited to chondrocyte, osteoblast, hepatocyte, fibroblast or smooth muscle cell in above-mentioned three kinds of technical solutions of the present invention.
The present invention combines the advantage of porous support and hydrogel support, both keep the favorable mechanical performance of porous support, macroscopical plastotype performance and macroshape hold facility, had the good imitative extracellular matrix structure and the bio-compatible performance of hydrogel support again.By changing the component of colloidal sol, for different cells, can add various promoting growth of cell and differentiation factor targetedly, thereby further improve the biology performance of support, method is simply effective.The present invention is applied widely, is applicable to the porous support and the hydrogel of multiple structure and composition, and various kinds of cell; Preparation technology is simple, good reproducibility, and prepared support can be widely used in field of tissue engineering technology.The present invention provides a kind of practicable method for the best combination of cell in the Tissue Engineering Study and support.Porous support by the present invention preparation can be widely used in the reconstruction and the regeneration of multiple tissue such as skin, cartilage, bone, liver, blood vessel, trachea, esophagus, tendon, cardiac valve or organ.
Description of drawings
Fig. 1 is the distribution situation of chondrocyte in the compound polylactic acid bracket of sodium alginate gel;
Fig. 2 is the distribution situation of chondrocyte in the compound polylactic acid bracket of agar gel;
Fig. 3 is the distribution situation of chondrocyte in agar and the compound polylactic acid bracket of gelatin gel;
Fig. 4 is the distribution situation of chondrocyte in basic fibroblast growth factor and the compound polylactic acid bracket of agar gel;
Fig. 5 is the distribution situation of osteoblast in acid fibroblast growth factor and sodium alginate gel composite collagen support;
Fig. 6 a is the laser confocal microscope photo that 100 microns polylactic acid porous scaffolds (black part) are filled sodium alginate hydrogel (white portion);
Fig. 6 b is the laser confocal microscope photo that 300 microns polylactic acid porous scaffolds (black part) are filled sodium alginate hydrogel (white portion);
Fig. 7 a is the laser confocal microscope photo (rhodamine dyeing) of collagen porous support;
Fig. 7 b is the laser confocal microscope photo (fluorescein sodium dyeing) that is distributed in the sodium alginate in the collagen porous support;
Fig. 7 c is the composite diagram of Fig. 6 a and Fig. 6 b.
The specific embodiment
Following example further specifies the present invention, but these examples are not used for limiting the present invention.
Configuration bulking value concentration is 2% sodium alginate colloidal sol, puts into 120 ℃ of high temperature sterilizes of autoclave 30 minutes, be cooled to room temperature after, sodium alginate is mixed with the chondrocyte suspension, the concentration that makes final sodium alginate is 1%.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.The polylactic acid bracket surface after processing with colloidal sol and cell mixture dropping.Under gravity and capillarity, colloidal sol enters internal stent.To prop up then and be placed in 0.5% the calcium lactate solution 15 minutes, by the crosslinked sodium alginate gelization that makes of calcium ion.At last, compound rest is placed 5%CO
2, hatched 7 days in 37 ℃ of homothermic incubators, the survival of test cell and extracellular matrix secretion situation, and compare with the blank polylactic acid bracket that does not import the sodium alginate hydrogel see Table 1.The form of cell and distribution situation are seen Fig. 1 in the support.As seen chondrocyte can be evenly distributed in the polylactic acid bracket that is compounded with the sodium alginate hydrogel, and cell activity is higher.
Import the sodium alginate hydrogel in table 1 polylactic acid porous scaffold to the chondrocyte Effect on Performance
| Support | The MTT activity | Hydroxyproline (mg/g) | GAG(mg/g) |
| Blank polylactic acid bracket | 100 | 6.22 | 11.47 |
| The compound polylactic acid bracket of sodium alginate | 146.9 | 6.60 | 11.11 |
MTT refers to that (cytoactive of measurement is 100 to carry out normalized with blank polylactic acid bracket for 3-(4, the 5-dimethylthiazole)-2,5-diphenyl tetrazole bromine salt (tetrazolium bromide).GAG represents chondroitin sulfate.The unit of hydroxyproline and GAG is the milligram number of emiocytosis in every gram support.
Embodiment 2 polylactic acid porous scaffolds import the agar hydrogel that contains chondrocyte
Configuration bulking value concentration is 2% agar colloidal sol, puts into 120 ℃ of high temperature sterilizes of autoclave 30 minutes, be cooled to about 45 ℃ after, agar colloidal sol is mixed with the chondrocyte suspension, the concentration that makes final agar is 1%.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.The polylactic acid bracket surface of mixture dropping after processing with colloidal sol and cell.Under gravity and capillarity, colloidal sol enters internal stent.Then support is placed down at 25 ℃ and made agar gelization in 15 minutes.At last, compound rest is placed 5%CO
2, hatched 7 days in 37 ℃ of homothermic incubators, the survival of test cell and extracellular matrix secretion situation, and compare with the blank polylactic acid bracket that does not import the agar hydrogel see Table 2.The form of cell and distribution situation are seen Fig. 2 in the support.As seen chondrocyte can be evenly distributed in the polylactic acid bracket that is compounded with the agar hydrogel, and cell activity is higher, and the secretory volume of hydroxyproline and GAG is bigger.
Import the agar hydrogel in table 2 polylactic acid porous scaffold to the chondrocyte Effect on Performance
| Support | The MTT activity | Hydroxyproline (mg/g) | GAG(mg/g) |
| Blank polylactic acid bracket | 100 | 6.22 | 11.47 |
| The compound polylactic acid bracket of agar | 201.5 | 9.79 | 67.25 |
Embodiment 3 polylactic acid porous scaffolds import agar and the gelatin hydrogel that contains chondrocyte
Configuration bulking value concentration respectively is 2% agar, gelatin mixed sols, put into 120 ℃ of high temperature sterilizes of autoclave 30 minutes, after being cooled to about 45 ℃, agar colloidal sol, gelatin colloidal sol are mixed with the chondrocyte suspension, make that the concentration of final agar and gelatin respectively is 1%.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.The polylactic acid bracket surface of mixture dropping after processing with colloidal sol and cell.Under gravity and capillarity, colloidal sol enters internal stent.Then support is placed down at 25 ℃ and made agar gelization in 15 minutes.At last, compound rest is placed 5%CO
2, hatched 7 days in 37 ℃ of homothermic incubators, the survival of test cell and extracellular matrix secretion situation, and compare with blank polylactic acid bracket that does not import the agar hydrogel and the simple polylactic acid bracket that imports agar see Table 3.The form of cell and distribution situation are seen Fig. 3 in the support.As seen chondrocyte can be evenly distributed in the polylactic acid bracket that is compounded with agar and gelatin hydrogel, and cell activity is higher, and the secretory volume of hydroxyproline and GAG is bigger.
Import agar and gelatin hydrogel in table 3 polylactic acid porous scaffold to the chondrocyte Effect on Performance
| Support | The MTT activity | Hydroxyproline (mg/g) | GAG(mg/g) |
| Blank polylactic acid bracket | 100 | 6.22 | 11.47 |
| The compound polylactic acid bracket of agar | 201.5 | 9.79 | 67.25 |
| Agar and gelatin-compounded polylactic acid bracket | 264.5 | -- | 77.62 |
Because gelatin molecule contains a large amount of hydroxyprolines, thus add can't distinguish behind the gelatin hydroxyproline be emiocytosis or gelatin in.
Embodiment 4 polylactic acid porous scaffolds import and contain chondrocyte and the alkaline agar hydrogel of facilitating fibroblast growth factor
Configuration bulking value concentration is 2% agar colloidal sol, put into 120 ℃ of high temperature sterilizes of autoclave 30 minutes, after being cooled to about 45 ℃, agar colloidal sol and basic fibroblast growth factor (bFGF) are mixed with the chondrocyte suspension, the concentration that makes final agar is 1%, and bFGF concentration is 100 μ g/mL.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.The polylactic acid bracket surface of mixture dropping after processing with colloidal sol and cell.Under gravity and capillarity, colloidal sol enters internal stent.Then support is placed down at 25 ℃ and made agar gelization in 15 minutes.At last, compound rest is placed 5%CO
2, hatched 7 days in 37 ℃ of homothermic incubators, the survival of test cell and extracellular matrix secretion situation, and compare with blank polylactic acid bracket and the simple polylactic acid bracket that imports the agar hydrogel see Table 4.The form of cell and distribution situation are seen Fig. 4 in the support.As seen chondrocyte can be evenly distributed in the polylactic acid bracket that is compounded with the agar hydrogel, and cell activity has the raising of highly significant, and the secretory volume of hydroxyproline and GAG is filled with simple agar does not have significantly difference.
Import bFGF and agar water-setting chamber in table 4 polylactic acid porous scaffold to the chondrocyte Effect on Performance
| Support | The MTT activity | Hydroxyproline (mg/g) | GAG(mg/g) |
| Blank polylactic acid bracket | 100 | 6.22 | 11.47 |
| The compound polylactic acid bracket of agar | 201.5 | 9.79 | 67.25 |
| The compound polylactic acid bracket of agar and bFGF | 370.1 | 9.53 | 55.90 |
Embodiment 5 collagen porous supports import and contain osteoblast and the acid agar hydrogel of facilitating fibroblast growth factor
Configuration bulking value concentration is 2% sodium alginate colloidal sol, put into 120 ℃ of high temperature sterilizes of autoclave 30 minutes, after being cooled to room temperature, facilitate fibroblast growth factor mixing (aFGF) to mix sodium alginate and acidity with the osteoblast suspension, the concentration that makes final sodium alginate is 1%, and aFGF concentration is 30 μ g/mL.Average pore size is about 100 microns, and porosity is 99.9%, and thick about 2 millimeters collagen thin slice support places tri-distilled water to soak, and makes the support swelling.Afterwards support is immersed in the mixed liquor of colloidal sol and cell and shake half an hour, make sodium alginate and chondrocyte enter collagen scaffold inside.To prop up then and be placed in 0.1% the calcium lactate solution 10 minutes, by the crosslinked sodium alginate gelization that makes of calcium ion.At last, compound rest is placed 5%CO
2, hatched 7 days in 37 ℃ of homothermic incubators, the form of cell and distribution situation are seen Fig. 5 in the support.
Embodiment 6 polylactic acid porous scaffolds import the sodium alginate hydrogel
At room temperature pre-configured bulking value concentration is 1%, 2% sodium alginate colloidal sol.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.With the polylactic acid bracket surface of sodium alginate colloidal sol dropping after processing.Under gravity and capillarity, colloidal sol enters internal stent.To prop up then and be placed in 0.5% the calcium lactate solution 60 minutes, by the crosslinked sodium alginate gelization that makes of calcium ion.After rack surface cleaned with tri-distilled water, drying was weighed.Utilize following formula to calculate the importing rate:
Importing rate=actual bracket weightening finish/Theoretical Calculation support weightening finish
Found that in sodium alginate hydrogel concentration to be respectively 1% and at 2% o'clock, the importing rate is respectively 92% and 94%.
Embodiment 7 polylactic acid porous scaffolds import the agar hydrogel
At room temperature pre-configured bulking value concentration is 1%, 2% agar colloidal sol.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.With concentration is that 1% or 2% agar colloidal sol drips the polylactic acid bracket surface after processing.Under gravity and capillarity, colloidal sol enters internal stent.Then support is placed down at 10 ℃ and made agar gelization in 60 minutes.After the rack surface of importing hydrogel cleaned with tri-distilled water, drying was weighed, and calculates the importing rate.Found that in agar hydrogel concentration to be respectively 1% and at 2% o'clock, the importing rate is respectively 100% and 83%.
Embodiment 8 polylactic acid porous scaffolds import aquagel
200mg chitosan (removing the acetyl degree is 91%) is dissolved in the 9ml 0.1M hydrochloric acid solution.560mg phosphoglyceride disodium is dissolved in the 1ml deionized water, and solution dropwise adds in the chitosan solution, forms homogeneous solution.With average pore size is 300 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.The support of handling is dipped in the aquagel, handled 5 minutes at 37 ℃ after 20 minutes, make chitosan gel rubberization.After the rack surface of importing hydrogel cleaned with tri-distilled water, drying was weighed, and recording the importing rate is 84.6%.
Implement of the influence of 9 polylactic acid porous scaffold apertures to hydrogel importing rate
At room temperature pre-configured bulking value concentration is 2% sodium alginate colloidal sol.With average pore size is 300 microns and 100 microns, and thickness is the polylactic acid bracket thin slice of 2-4 millimeter, places 75% alcoholic solution to soak, and by the way of evacuation, the air in the support is removed, and makes alcoholic solution soak into support fully.Soak in alcoholic solution after one day, it is that 7.4 phosphate buffer soaked 1 day that support is put into pH, during change liquid 3 times, to displace the ethanol in the support fully.Sodium alginate colloidal sol is mixed with fluorescein sodium, and the concentration that makes final sodium alginate is 1%, and the concentration of fluorescein sodium is 0.01%, and drips the polylactic acid bracket surface after processing.Under gravity and capillarity, colloidal sol enters internal stent.To prop up then and be placed in 2% the calcium lactate solution 5 minutes, by the crosslinked sodium alginate gelization that makes of calcium ion.Adopt laser confocal microscope to observe hydrogel distribution situation therein, see Fig. 6 a, Fig. 6 b.After rack surface cleaned with tri-distilled water, drying was weighed.Found that in the aperture of polylactic acid porous scaffold the importing rate is respectively 82% and 92% when being 100 microns and 300 microns.Illustrate that the large aperture helps the importing of hydrogel.
Embodiment 10 collagen porous supports import the sodium alginate hydrogel
Average pore size is about 100 microns, and porosity is 99.9%, and thick about 2 millimeters rhodamine labelling collagen thin slice support places tri-distilled water to soak, and makes the support swelling.Again support is immersed the concentration be dissolved with 0.01% fluorescein sodium and be in 1% or 2% the sodium alginate colloidal sol and shake half an hour, make sodium alginate colloidal sol enter collagen scaffold inside.To prop up then and be placed in 0.1% the calcium lactate solution 50 minutes, by the crosslinked sodium alginate gelization that makes of calcium ion.The employing laser confocal microscope is observed, and sees Fig. 7 a, Fig. 7 b, Fig. 7 c.After rack surface cleaned with tri-distilled water, drying was weighed.Found that in sodium alginate hydrogel concentration to be respectively 1% and at 2% o'clock, the importing rate is respectively 100% and 84%.
Claims (10)
1. hydrogel is compound to the method in the porous tissue engineering scaffold, this method may further comprise the steps:
1) under 4 ℃～120 ℃ agar is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the colloidal sol bulking value concentration that obtains is 0.5～5%;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the hydrogel colloidal sol, hydrogel is adsorbed onto porous support inside;
4) support that will be compounded with colloidal sol was placed 10 minutes under 15C～40C at least, the porous support of hydrogel that promptly obtained load.
2. according to claim 1 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that in step 1), in agar, sneak into gelatin or/and cell growth factor, the ratio of gelatin weight and agar weight is 1: 1～1: 100, the ratio of cell growth factor weight and agar weight is 1: 10,000～1: 1,000,000.
3 according to claim 2ly are compound to method in the porous tissue engineering scaffold with hydrogel, it is characterized in that said cell growth factor is that alkalescence facilitates fibroblast growth factor, acidity to facilitate fibroblast growth factor, bone morphogenetic protein(BMP) or transforming factor β.
4. according to claim 1 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that said porous support is the polymer class porous support, comprise polylactic acid, collagen, lactic acid-ethanol copolymer, polycaprolactone, polyurethane or polystyrene holder, or be the ceramic-like porous support, comprise porous hydroxyapatite, tricalcium phosphate or bio-vitric support; Said cell is chondrocyte, osteoblast, hepatocyte, fibroblast or smooth muscle cell.
5. hydrogel is compound to the method in the porous tissue engineering scaffold, this method may further comprise the steps:
1) under 4 ℃～120 ℃ sodium alginate is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the colloidal sol bulking value concentration that obtains is 0.5～5%;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the hydrogel colloidal sol, hydrogel is adsorbed onto porous support inside;
4) will be compounded be placed in 0.1%～2% calcium lactate solution at least 5 minutes of colloidal sol, the porous support of hydrogel that promptly obtained load.
6. according to claim 5 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that in step 1), in sodium alginate, sneak into gelatin or/and cell growth factor, the ratio of gelatin weight and sodium alginate weight is 1: 1～1: 100, the ratio of cell growth factor weight and sodium alginate weight is 1: 10,000～1: 1,000,000.
7. according to claim 6 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that described cell growth factor is that alkalescence facilitates fibroblast growth factor, acidity to facilitate fibroblast growth factor, bone morphogenetic protein(BMP) or transforming factor β.
8. according to claim 5 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that said porous support is the polymer class porous support, comprise polylactic acid, collagen, lactic acid-ethanol copolymer, polycaprolactone, polyurethane or polystyrene holder, or be the ceramic-like porous support, comprise porous hydroxyapatite, tricalcium phosphate or bio-vitric support; Said cell is chondrocyte, osteoblast, hepatocyte, fibroblast or smooth muscle cell.
9. hydrogel is compound to the method in the porous tissue engineering scaffold, this method may further comprise the steps:
1) under 4 ℃～120 ℃ with the phosphoglyceride disodium salt mixture of chitosan and 1～5 times of weight, or polyoxyethylene-polyoxypropylene copolymer, or PNIPAM is dissolved in the water and is prepared into the colloidal sol of hydrogel, mix with isopyknic celliferous culture medium solution, the bulking value concentration that obtains colloidal sol is 0.5～5% again;
2) porous support is carried out sterilization treatment, the moistening in advance of reuse phosphate buffer;
3) colloidal sol is added drop-wise to the surface of porous support, under gravity and capillarity, colloidal sol is imported internal stent; Or support is immersed in the colloidal sol of hydrogel, hydrogel is adsorbed onto porous support inside;
4) will be compounded be placed under 30 ℃～40 ℃ conditions at least 5 minutes of colloidal sol, the porous support of hydrogel that promptly obtained load.
10. according to claim 9 hydrogel is compound to method in the porous tissue engineering scaffold, it is characterized in that said porous support is the polymer class porous support, comprise polylactic acid, collagen, lactic acid-ethanol copolymer, polycaprolactone, polyurethane or polystyrene holder, or be the ceramic-like porous support, comprise porous hydroxyapatite, tricalcium phosphate or bio-vitric support; Said cell is chondrocyte, osteoblast, hepatocyte, fibroblast or smooth muscle cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100529844A CN1314460C (en) | 2004-07-15 | 2004-07-15 | Method for compounding aquo-gel to porous tissue engineering rack |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100529844A CN1314460C (en) | 2004-07-15 | 2004-07-15 | Method for compounding aquo-gel to porous tissue engineering rack |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1586636A CN1586636A (en) | 2005-03-02 |
| CN1314460C true CN1314460C (en) | 2007-05-09 |
Family
ID=34602687
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100529844A Expired - Fee Related CN1314460C (en) | 2004-07-15 | 2004-07-15 | Method for compounding aquo-gel to porous tissue engineering rack |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1314460C (en) |
Families Citing this family (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100431624C (en) * | 2005-12-07 | 2008-11-12 | 浙江大学 | Method for preparing injectable polyletic acid micro-carrier/chitosan hydrogel composite scaffold |
| WO2007092043A2 (en) * | 2006-02-02 | 2007-08-16 | Medlogics Device Corporation | Bioactive material delivery systems comprising sol-gel compositions |
| CN101396569B (en) * | 2007-09-27 | 2013-04-10 | 北京师范大学 | Composite hydrogel using organism tissue as bracket, preparation method and use thereof |
| CN103203210A (en) * | 2008-03-27 | 2013-07-17 | 哈佛学院院长等 | Paper-based Cellular Arrays |
| CN101829361B (en) * | 2009-03-10 | 2013-08-21 | 广州迈普再生医学科技有限公司 | Nano-bionic material for tissue repair and preparation method thereof |
| CN101574543B (en) * | 2009-06-09 | 2014-10-29 | 广州迈普再生医学科技有限公司 | Artificial articular cartilage based on autologous cells and preparation method thereof |
| CN101716382B (en) * | 2009-12-02 | 2015-07-15 | 浙江大学 | Preparation method of trinary composite stent of plasmid DNA / fibrin gel / polymer |
| CN102198022B (en) * | 2011-05-23 | 2013-04-17 | 西安交通大学 | Solid forming method of active cell-hydrogel organ structure |
| CN102440852A (en) * | 2011-12-07 | 2012-05-09 | 上海交通大学 | Mixed porous structure interbody fusion cage and preparation method thereof |
| CN102526810B (en) * | 2012-02-22 | 2014-04-09 | 四川大学 | Artificial skin alternative material and preparation method thereof |
| CN103893818B (en) * | 2014-03-14 | 2015-07-01 | 华南理工大学 | Osteochondral three-dimensional stent with regular interpenetrating network structure and preparation method thereof |
| CN104667357B (en) * | 2015-02-05 | 2017-12-22 | 中国科学院兰州化学物理研究所 | A kind of composite lubricated material of structured hydrogel and preparation method thereof |
| CN104940996A (en) * | 2015-06-04 | 2015-09-30 | 浙江大学 | Preparation method of radial collagen scaffold in combination with stromal cell derived factor and application of radial collagen scaffold in combination with stromal cell derived factor to osteochondral defect repair |
| CN105288737B (en) * | 2015-09-30 | 2018-08-07 | 中国人民解放军总医院 | A kind of tissue engineering bone/cartilage compound rest and preparation method thereof |
| CN106730026B (en) * | 2017-03-01 | 2022-05-27 | 北京大学第三医院 | Tissue engineering cartilage composite scaffold and preparation method thereof |
| CN106668948B (en) * | 2017-03-01 | 2018-02-06 | 北京大学第三医院 | A kind of tissue engineering bracket and preparation method based on low temperature rapid shaping |
| WO2018183998A1 (en) * | 2017-03-31 | 2018-10-04 | Cellum Biomedical, Inc. | Biocompatible conditioned cell medium compositions and uses thereof |
| CN107617126A (en) * | 2017-08-11 | 2018-01-23 | 天津大学 | A kind of surface modification method of porous material and application |
| CN107715175B (en) * | 2017-09-30 | 2018-10-23 | 南京师范大学 | A kind of compound porous bone tissue engineering scaffold of PLGA/ amphoteric ions and preparation method thereof |
| CN108452388A (en) * | 2018-01-15 | 2018-08-28 | 朱向情 | A kind of cell-scaffold composite material and preparation method thereof and its application for bone defect healing and reparation |
| CN108853578A (en) * | 2018-06-15 | 2018-11-23 | 南京冬尚生物科技有限公司 | A kind of 3D printing Ti- hydrogel-osteoblast bone tissue engineering scaffold and preparation method thereof |
| CN109045352A (en) * | 2018-09-03 | 2018-12-21 | 南京冬尚生物科技有限公司 | A kind of 3D printing Ti- hydrogel-vancomycin Anti-infective bone bracket and preparation method and application |
| CN110101903B (en) * | 2019-04-09 | 2021-11-02 | 温州医科大学 | BG composite scaffold capable of inducing hypoxia and application thereof |
| CN111558086A (en) * | 2020-04-29 | 2020-08-21 | 广州市红十字会医院（暨南大学医学院附属广州红十字会医院） | Cartilage repair material containing translation regulation tumor protein and preparation method and application thereof |
| CN113349988A (en) * | 2021-05-31 | 2021-09-07 | 浙江大学 | Tissue engineering bone for repairing jaw cleft palate defect and preparation method thereof |
| CN114457018A (en) * | 2022-02-17 | 2022-05-10 | 安徽骆华生物科技有限公司 | Three-dimensional breast cancer organoid model and culture method and application thereof |
| CN115105630B (en) * | 2022-08-08 | 2023-08-18 | 池州学院 | 3D printing material embedded with chitosan/gelatin composite hydrogel and preparation method thereof |
| CN115369065A (en) * | 2022-08-09 | 2022-11-22 | 华南理工大学 | Three-dimensional cell culture scaffold based on hydrogel and preparation method thereof |
| CN115252891A (en) * | 2022-08-26 | 2022-11-01 | 四川大学 | Preparation method of calcium phosphate ceramic for drug loading and drug loading method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310027A (en) * | 2001-02-06 | 2001-08-29 | 中国人民解放军第三军医大学第三附属医院 | Preparation of compound artificial skin |
| CN1392253A (en) * | 2002-07-04 | 2003-01-22 | 中国人民解放军第三军医大学第一附属医院 | Method of utilizing three-dimensional rack seed cell double phase inoculation to construct engineered tissue |
| WO2003043674A1 (en) * | 2001-11-16 | 2003-05-30 | Children's Medical Center Corporation ¨ | Augmentation of organ function |
| CN1422672A (en) * | 2002-11-21 | 2003-06-11 | 上海第二医科大学附属第九人民医院 | Absorbable calcined-bone preparation method |
-
2004
- 2004-07-15 CN CNB2004100529844A patent/CN1314460C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310027A (en) * | 2001-02-06 | 2001-08-29 | 中国人民解放军第三军医大学第三附属医院 | Preparation of compound artificial skin |
| WO2003043674A1 (en) * | 2001-11-16 | 2003-05-30 | Children's Medical Center Corporation ¨ | Augmentation of organ function |
| CN1392253A (en) * | 2002-07-04 | 2003-01-22 | 中国人民解放军第三军医大学第一附属医院 | Method of utilizing three-dimensional rack seed cell double phase inoculation to construct engineered tissue |
| CN1422672A (en) * | 2002-11-21 | 2003-06-11 | 上海第二医科大学附属第九人民医院 | Absorbable calcined-bone preparation method |
Non-Patent Citations (1)
| Title |
|---|
| 组织工程人工血管支架的预构 潘勇等，中华整形外科杂志，第19卷第1期 2003 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1586636A (en) | 2005-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1314460C (en) | Method for compounding aquo-gel to porous tissue engineering rack | |
| CN108084461B (en) | Controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and preparation method and application thereof | |
| CN102688525B (en) | Bio-macromolecular hydrogel and preparation method thereof | |
| US7871638B2 (en) | Composite material containing a calcium phosphate gradient | |
| CN105688274B (en) | A kind of preparation process of polycaprolactone/gelatin electrospinning compound rest | |
| CN108310467A (en) | A kind of packaging cell-derived extracellular matrix membrane composite bone repairing material and its preparation method and application | |
| CN107205809A (en) | The biological mixture regenerated for nerve tract | |
| CN111658820A (en) | Injectable hydrogel for promoting bone regeneration and preparation method thereof | |
| CN114712566B (en) | PCL/GelMA biological porous scaffold based on Pickering emulsion and preparation method thereof | |
| CN1411471A (en) | Macroporous chitosan beads and preparation method thereof | |
| Fu et al. | Sericin/nano-hydroxyapatite hydrogels based on graphene oxide for effective bone regeneration via immunomodulation and osteoinduction | |
| CN112972776B (en) | Multi-stage hole bone repair biological scaffold material with photothermal effect and preparation method thereof | |
| CN114392391A (en) | Additive manufactured porous polylactic acid scaffold, and biological activity improvement method and application thereof | |
| CN112076350B (en) | Biomimetic mineralized hydrogel with nano-micron composite structure and high mineral density as well as preparation method and application thereof | |
| WO2023179544A1 (en) | 3d-printed bone defect repair scaffold loaded with mesenchymal stem cell extracellular matrixes and preparation method therefor | |
| Chaochai et al. | Preparation of chitosan-gelatin based sponge cross-linked with GlcNAc for bone tissue engineering | |
| CN112370571A (en) | Bone tissue engineering scaffold material for treating jaw and face bone defect and synthesis method thereof | |
| CN107412860A (en) | A kind of preparation method based on hydroxyapatite compound support frame material | |
| KR20210105804A (en) | A composition for tissue engineering comprising Alginate, Gelatin and Cellulose Nanocrystals and use of the same | |
| CN112354016A (en) | Bionic artificial bone material and production method thereof | |
| KR20200112256A (en) | Method for manufacturing bone-induction bilayer membrane containing electrospun polycaprolactone/gelatin/β-TCP mat and alginate/gelatin hydrogel | |
| CN102671238A (en) | High-emulation tissue-engineered nerve repair material | |
| CN114870071B (en) | Silicon-based bioactive ink, natural inorganic silicon-based material flexible three-dimensional porous support and application | |
| RU2819701C1 (en) | Hydrogel biocomposite based on bacterial polysaccharides for use in tissue engineering | |
| CN114870082B (en) | High-strength composite collagen membrane and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070509 Termination date: 20140715 |
|
| EXPY | Termination of patent right or utility model |