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Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere

A morphogenetic protein and fibrin glue technology, applied in the field of medical materials, can solve the problems of inconvenient operation, loss, difficult to grasp the coagulation time, etc., and achieve the effect of reducing trauma and accelerating the healing process.

Inactive Publication Date: 2012-10-31
姚琦 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in clinical operations, there are often the following problems: when dissolving with normal saline, the active ingredient is easy to absorb and lose, and cannot maintain a high concentration in the local area; while using venous blood or bone marrow blood to dissolve, it is difficult to control the coagulation time. The blood has coagulated in vitro, loses injectability, and is inconvenient to operate, so a slow-release carrier that can control the coagulation time is also needed

Method used

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  • Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere
  • Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere
  • Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere

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Experimental program
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Effect test

specific Embodiment 1

[0038] 10 mg of rhBMP-2 dry powder was dissolved in 0.5 ml of 4M guanidine hydrochloride solution, filtered, and then the obtained guanidine hydrochloride solution was diluted into 10 mM pbs phosphate buffer solution to form a solution of about 10 mg / ml. And add 1% bovine serum albumin 1mg as protective agent, mix well to form inner water phase.

[0039] 100mg of PLGA was dissolved in 0.7ml of dichloromethane containing 2.5% Span-20 to form an oily phase,

[0040] 1ml of the inner water phase was added to 0.7ml of the oil phase for ultrasonic emulsification in an ice bath to form colostrum.

[0041] Add 1.7ml of colostrum to 10ml of the external aqueous phase containing 1% polyvinyl alcohol PVA, 0.5% Tween-20,

[0042] Stir at 800r / min for 4h to evaporate the solvent, filter, wash with distilled water, vacuum freeze-dry, and store at 4-8°C.

[0043] Scanning electron microscope observation shows that the microspheres are regular and uniform in shape, approximately round, wit...

specific Embodiment 2

[0046] From figure 2It can be seen from the figure that the release of rhBMP-2 from various materials is biphasic, and the rhBMP-2 / PLGA microspheres burst into release in the first 4 days, and the drug release exceeds 27.005%, mainly due to the rhBMP-2 adsorbed on the surface of the microspheres. The amount of rhBMP-2 released in this period was 3.65%, 5.17%, 8.95%, and 11.76% in 2h, 8h, 24h, and 2d, respectively. The subsequent 3-42d showed a sustained and slow release, and the amount of rhBMP-2 released reached 91.75% in 42 days.

[0047] rhBMP-2 / PLGA microspheres / fibrin glue released 1.15%, 1.75%, and 6.68% of BMP at 2h, 8h, and 24h, respectively. At the same time point were lower than rhBMP-2 / PLGA microsphere group. The release of 2dBMP reached 16.76%, and the subsequent 3-42d also showed a sustained and slow release, but the release was less than that of the rhBMP-2 / PLGA microsphere group, and the release of rhBMP-2 reached 76.75% in 42 days. This method is conducive ...

specific Embodiment 3

[0048] Such as image 3 Shown: the carbon element accounts for 71.26% in the PLGA microsphere, the oxygen element accounts for 28.74%, and no nitrogen element is found. Most of the polypeptide chains of fibrin glue are composed of C, N, and O elements, of which carbon elements account for 67.68%, nitrogen elements account for 8.34%, and oxygen elements account for 23.98%. After fibrin glue is compounded with microspheres, the carbon element increases to 68.54 %, oxygen element 27.02%, however nitrogen element decreased to 4.44%. Non-polar groups (C-C and C-H) in fibrin glue are 41.88%, wherein O=C-NH group is 36.21%, without C-NH2 and COOH (possibly two kinds of groups are less in composition in fibrin glue ), the polar group is 21.91%. After the microspheres were mixed into the fibrin glue, the groups changed C-C / C-H (52.61%), C-OH (33.39%) and O=C-NH (14.00%). Through energy spectrum analysis, we found that the microspheres were mixed into the fibrin The release process i...

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Abstract

The invention relates to a preparation method and application of a fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere. The preparation method comprises the steps of: preparing a slow release microsphere with a proper particle diameter; and then constructing an rhBMP-2 / PLGA microsphere / fibrin glue composite material. The rhBMP-2 / PLGA microsphere fibrin glue composite material can be used through local injection, surgical trauma can be reduced, a healing process of bone fracture and nonunion is accelerated by continuously supplementing the local bone morphogenetic protein, thus the fibrin glue composite recombinant human bone morphogenetic protein-2 is a bone repair material with excellent degradability and osteogenic activity.

Description

technical field [0001] The patent of the invention belongs to medical materials, mainly through local injection application, reducing surgical trauma, and continuously replenishing local bone morphogenetic protein 2 to promote osteogenesis and accelerate the healing process of fractures and nonunions. Background technique [0002] Fracture healing is an extremely complicated process of bone tissue regeneration and repair. Many factors can affect fracture healing, leading to delayed union or nonunion of fractures (nonunion). Researchers as well as clinicians have been working on treatments to promote fracture healing and reduce delayed union or nonunion of fractures. [0003] In 1965, American doctor Urist (Urist MR. Bone: formation byautoinduction. Science, 1965, 150 (698): 893-899) found that decalcified bone mesenchyme had ectopic osteogenesis, and the newly discovered ability to induce new bone The formed protein is named bone morphogenetic protein (Bone Morphogenetic pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/54A61K38/18A61K47/34A61K47/42A61P19/08
Inventor 姚琦姜华唐佩福
Owner 姚琦
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