CN1463755A - Method for preparation of growth factor carrier for bone tissue conversion - Google Patents
Method for preparation of growth factor carrier for bone tissue conversion Download PDFInfo
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- CN1463755A CN1463755A CN 02120967 CN02120967A CN1463755A CN 1463755 A CN1463755 A CN 1463755A CN 02120967 CN02120967 CN 02120967 CN 02120967 A CN02120967 A CN 02120967A CN 1463755 A CN1463755 A CN 1463755A
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Abstract
The preparation process of bone tissue converting growth factor carrier includes making hollow cylinder as the carrier, mixing bone morphogenetic protein, colloidal carrier, etc in certain proportion to form mixture, setting the mixture into the holes of the cylinder carrier and drying to form. The present invention has the advantage that bone morphogenetic protein is released from the center of the carrier and bone formation begins from the center to avoid the excessive release and degradation of bone morphogenetic protein and this is favorable to bone formation. The complete bone formation both outside and inside the carrier is favorable to the degradation of the carrier.
Description
Affiliated technical field:
The present invention relates to medical science bone implantation technique field, particularly relate to a kind of osseous tissue that is used for and transform the preparation method of giving birth to factor carrier.
Background technology:
In bone implantation technique field, the osseous tissue growth situation that implants is by the decision of its transforming growth factor that has, and shows bone morphogenetic protein(BMP) (BMP) in transforming growth factor (TGF-β) family etc. in clinical research, has the osteogenesis of inducing effect.After but simple bone morphogenetic protein(BMP) etc. implanted, very fast diffusion or diluted and enzymolysis absorption were difficult to play one's part to the full in new osteoplastic overall process; Therefore in addition, damaged for bigger bone, no support effect such as bone morphogenetic protein(BMP) needs suitable carriers, and bone morphogenetic protein(BMP) etc. is slowly discharged in the effect of performance support.
At present the carriers of using such as bone morphogenetic protein(BMP) have two kinds of physical aspects: 1. colloidal carrier, as collagen, fibrin clot (FC), polypyrrole alkane ketone etc.; 2. solid-state carrier, common as: hydroxyapatite (hydroxyapatite, HA), polylactic acid (polylactic acid, PLA), macrogol (polyethyleneglycol, PEG), polyglycolic acid (polygiycolic acid, PGA) and their copolymer; Sintering bone, allogeneic decalcification bone or demineralized bone matrix, freeze drying bone, deproteinization bone, reconstituted bone xenograft and Corallium Japonicum Kishinouye bone etc.; 3. be to mix by two kinds of form carriers to use in addition.
The complex method of colloidal carrier and bone morphogenetic protein(BMP) etc. is: directly with bone morphogenetic protein(BMP) etc. and carrier mixing; The complex method of solid-state carrier and bone morphogenetic protein(BMP) etc. is: carrier is dipped in the solution that contains bone morphogenetic protein(BMP) etc., by negative pressure-pumping bone morphogenetic protein(BMP) is waited until in the carrier; Combination carrier and the complex method of bone morphogenetic protein(BMP) then are earlier bone morphogenetic protein(BMP) etc. to be mixed with colloidal carrier, this liquid are adsorbed onto in the solid-state carrier by negative pressure-pumping again.
These several bearing modes have some unfavorable: one, bone morphogenetic protein(BMP) etc. distributes unevenly, and it is many and inner few that the carrier outside contains bone morphogenetic protein(BMP) etc.The release imbalance that causes bone morphogenetic protein(BMP) etc. discharges manyly in early days, is organized the too fast degraded of protease, and that the later stage discharges is few, does not reach the inductive valid density of bone.Two, because the skewness of bone morphogenetic protein(BMP) etc. causes skeletonization mainly on the surface of carrier, and inner skeletonization is few, thereby skeletonization is not exclusively with slow.Three, the appointed condition requirement is high, production process is complicated.
Summary of the invention:
In order to solve the deficiency that exists in above-mentioned several bearing mode, the invention provides a kind of preparation method that is used for osseous tissue transforming growth factor carrier, this method is based in the carrier bone morphogenetic protein(BMP) by the center slow release, and skeletonization is by setting up in theory of beginning of center.Its advantage is: 1. bone morphogenetic protein(BMP) etc. discharges towards periphery from carrier center, has avoided bone morphogenetic protein(BMP) etc. to be discharged in a large number in the tissue and is degraded, and forms a lasting bone morphogenetic protein(BMP) isoconcentration gradient at carrier and periphery and helps skeletonization.The inside and outside skeletonization of carrier is complete, and then helps the degraded of carrier.2. equipment requirements is simple, and is simple and easy to do.
Concrete technical scheme of the present invention is; A kind ofly be used for the preparation method that osseous tissue transform to be given birth to factor carrier, be characterized in: all kinds of solid carriers are processed into the cylinder of hollow, and bone morphogenetic protein(BMP) etc. are inserted in the solid carrier mesopore drying and moulding after pressing the abundant mixing of special ratios with colloid sample carrier.
Concrete steps are:
1. allograph bone is made diameter 6-10mm, length 3-6mm, hollow, the cylinder of internal diameter 2-4mm.
With the cylinder for preparing with methanol/chloroform (1: 1) supersound washing 6h, 0.6 equivalent hydrochloric acid, decalcification in 15 minutes, 3% hydrogen peroxide 6h deproteinization, the antigen bone is removed in preparation.
3. collagen is mixed respectively with 1: 1,1: 2,1: 4,1: 8,1: 16 ratio with gelatin and add an amount of distilled water, the abundant mixing of vortex oscillator concussion 24h is regulated pH value 7.0, lyophilizing, preparation collagen gelatin mixture.
4. go antigen allograph bone and collagen gelatin complex to use oxirane disinfection standby respectively.
5. under the sterile working, quantitative collagen gelatin complex is mixed with bone morphogenetic protein(BMP), add a small amount of sterile distilled water and be mixed into glue, place antigen bone mesopore, drying.
After this kind complex implanted, because stopping of outer solid carrier arranged, the enzyme in the body fluid can not touch bone morphogenetic protein(BMP) at once, thereby has effectively preserved bone morphogenetic protein(BMP); Along with body fluid penetrates into mesopore, bone morphogenetic protein(BMP) is slowly discharged around carrier by the center gradually, forms bone morphogenetic protein(BMP) gradient concentration from high to low from the mesopore to the periphery in solid carrier, and then osteogenetic process is carried out towards periphery by the center.
Description of drawings:
Fig. 1 is packed into the solid carrier sketch map of colloid fiber complex of the present invention
The specific embodiment:
Present embodiment is with allograph bone (solid carrier), and the combination process of collagen and gelatin (colloidal carrier) and bone morphogenetic protein(BMP) is the example processor of light staining center release vehicle and bone morphogenetic protein compounded specifically.
Concrete steps are:
1. allograph bone is made diameter 8mm, length 5mm, hollow, the cylinder of internal diameter 3mm.
With the cylinder for preparing with methanol/chloroform (1: 1) supersound washing 6h, 0.6 equivalent hydrochloric acid, decalcification in 15 minutes, 3% hydrogen peroxide 6h deproteinization, the antigen bone is removed in preparation.
3. collagen is mixed respectively with 1: 1,1: 2,1: 4,1: 8,1: 16 ratio with gelatin and add an amount of distilled water, the abundant mixing of vortex oscillator concussion 24h is regulated pH value 7.0, lyophilizing, preparation collagen gelatin mixture.
4. go antigen allograph bone and collagen gelatin complex to use oxirane disinfection standby respectively.
5. under the sterile working, quantitative collagen gelatin complex is mixed with bone morphogenetic protein(BMP), add a small amount of sterile distilled water and be mixed into glue, place antigen bone mesopore, drying.
Above step is referring to accompanying drawing 1, and 1 is space property solid carrier among the figure; 2 for containing the colloid fiber complex of medicine.
This method is in cell culture experiments in vitro, and it is slow to find that the hollow bone discharges the more solid bone of bone morphogenetic protein(BMP), illustrates that the hollow structure of the present invention's design helps the reservation of bone morphogenetic protein(BMP).
In zoopery, with the bone morphogenetic protein(BMP)/collagen gelatin complex/allograph bone complex for preparing, allograph bone is not had hollow structure, and other contain complex with sample ingredient in contrast with allograph bone (solid carrier), each 5 in collagen and gelatin (colloidal carrier) and bone morphogenetic protein(BMP), implant rabbit spinal column both sides together, respectively at putting to death 2 all 4 samples, 4 all 6 samples 2,4 weeks.The result:
2 when week solid construction to discharge bone morphogenetic protein(BMP) more hollow fast, ALP is active high, but during to 4 weeks, hollow-core construction still slowly discharges bone morphogenetic protein(BMP), the activity of ALP is higher than solid construction, and the active suppression ratio of ALP is milder, and the release that bone morphogenetic protein(BMP) is described is slowly to continue.Do not resemble solid construction and discharge bone morphogenetic protein(BMP), in early days too, the later stage then is short of.
HE cut sections for microscopic examination 2 acellular growing in visible solid construction implant during week, and in the hollow-core construction implant a small amount of cartilage is arranged, the osteocyte growth.
Claims (2)
1. one kind is used for the preparation method that osseous tissue transform to be given birth to factor carrier, it is characterized in that: all kinds of solid carriers are processed into the cylinder of hollow, and bone morphogenetic protein(BMP) etc. are inserted in the solid carrier mesopore drying and moulding after pressing the abundant mixing of special ratios with colloid sample carrier.
Concrete steps are:
(1) allograph bone is made diameter 6-10mm, length 3-6mm, hollow, the cylinder of internal diameter 2-4mm.
(2) with cylinder methanol/chloroform (1: 1) the supersound washing 6h for preparing, 0.6 equivalent salt, decalcification in 15 minutes, 3% hydrogen peroxide 6h deproteinization, the antigen bone is removed in preparation.
(3) collagen is mixed the adding distilled water with gelatin respectively with 1: 1,1: 2,1: 4,1: 8,1: 16 ratio, the abundant mixing of vortex oscillator concussion 24h is regulated pH value 7.0, lyophilizing, preparation collagen gelatin mixture.
(4) go antigen allograph bone and collagen gelatin complex to use oxirane disinfection standby respectively.
(5) under the sterile working, quantitative collagen gelatin complex is mixed with bone morphogenetic protein(BMP), add a small amount of sterile distilled water and be mixed into glue, place antigen bone mesopore, drying.
2. preparation method according to claim 1, its feature with in: described all kinds of carriers comprise: hydroxyapatite (hydroxyapatite, HA), polylactic acid (polylactic acid, PLA), macrogol (polyethylene glycol, PEG), polyglycolic acid (polygiycolic acid, PGA) and their copolymer; Sintering bone, allogeneic decalcification bone or demineralized bone matrix, freeze drying bone, deproteinization bone, reconstituted bone xenograft and Corallium Japonicum Kishinouye bone etc.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02120967 CN1463755A (en) | 2002-06-06 | 2002-06-06 | Method for preparation of growth factor carrier for bone tissue conversion |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02120967 CN1463755A (en) | 2002-06-06 | 2002-06-06 | Method for preparation of growth factor carrier for bone tissue conversion |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101507831B (en) * | 2009-03-25 | 2012-12-26 | 北京大清生物技术有限公司 | Material for promoting operation wound heal and preparation method thereof |
| CN106413767A (en) * | 2014-04-25 | 2017-02-15 | 生物银行公司 | Method for producing a bone paste |
| CN106822997A (en) * | 2017-03-15 | 2017-06-13 | 杭州易文赛科拓干细胞技术研究有限公司 | A kind of preparation method for removing antigen bone holder material |
| WO2020207296A1 (en) * | 2019-04-11 | 2020-10-15 | 中南大学湘雅医院 | Apparatus for releasing animal intraosseous foreign particle |
-
2002
- 2002-06-06 CN CN 02120967 patent/CN1463755A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101507831B (en) * | 2009-03-25 | 2012-12-26 | 北京大清生物技术有限公司 | Material for promoting operation wound heal and preparation method thereof |
| CN106413767A (en) * | 2014-04-25 | 2017-02-15 | 生物银行公司 | Method for producing a bone paste |
| CN106413767B (en) * | 2014-04-25 | 2019-06-04 | 生物银行公司 | The method for producing bone mud |
| CN106822997A (en) * | 2017-03-15 | 2017-06-13 | 杭州易文赛科拓干细胞技术研究有限公司 | A kind of preparation method for removing antigen bone holder material |
| WO2020207296A1 (en) * | 2019-04-11 | 2020-10-15 | 中南大学湘雅医院 | Apparatus for releasing animal intraosseous foreign particle |
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