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Injected tissue engineering bone and its constrction and application

A tissue-engineered bone and injection-based technology, which is applied in bone diseases, drug combinations, medical preparations containing active ingredients, etc., can solve the problems of complex cell operation, expensive exogenous cytokines, etc., and achieve easy compounding and promotion. The effect of fracture healing and simple operation

Inactive Publication Date: 2006-10-18
NAN FANG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the above-mentioned prior art situation, in order to overcome the deficiencies of existing tissue-engineered bone such as difficulty in shaping, complicated operation of composite cells, and expensive exogenous cytokines, the present invention provides an injection-type tissue-engineered bone. The tissue-engineered bone is The injectable material is loaded with autologous PRP and then compounded with autologous bone marrow stromal cells. It can be shaped arbitrarily and the composite cells are simple, and autologous cell growth factors are used, so there is no immune rejection problem

Method used

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  • Injected tissue engineering bone and its constrction and application
  • Injected tissue engineering bone and its constrction and application
  • Injected tissue engineering bone and its constrction and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Culture and induction of differentiation of bone marrow stromal stem cells.

[0038] New Zealand white rabbits, provided by the Animal Institute of Nanfang Hospital, Southern Medical University (formerly First Military Medical University), were anesthetized by intramuscular injection of Sumianxin, ketamine and atropine compound solution. Extract 3ml of red bone marrow from bilateral iliac crests, anticoagulate with heparin solution, mix into DMEM medium, centrifuge at 1000r / min for 10min after mixing, discard the suspended fat cells and part of the supernatant, pass through a 100-mesh filter, and add DMEM complete medium (containing 15% fetal bovine serum) inoculation. 37°C, 5% CO 2 Culture in an incubator, change the full amount of medium after 3 days, and change the medium once in half of the next 2 to 3 days. After the cells cover 80% of the bottom of the bottle, digest with 0.25% trypsin, and subculture with DMEM conditioned medium (containing 10% fetal ...

Embodiment 2

[0039] Embodiment two: the extraction of PRP.

[0040] Under aseptic conditions, 5ml of central artery blood was extracted from the dorsal ear of rabbits, and anticoagulated with 0.5ml of 10% sodium citrate. Centrifuge at 3000r / min for the first time at 20°C for 10 minutes, and absorb the supernatant and 3 mm components below the buffy coat. Centrifuge for the second time at the same temperature at 3600r / min for 15 minutes, discard the upper 3 / 4 supernatant, and the remaining part is PRP. Vortex gently to mix. The platelet content was counted under a high power microscope and compared with whole blood. The number of platelets in the PRP constructed by this method is more than 4 times higher than that in whole blood. Make about 0.6ml of PRP per 5ml of whole blood, and store it in a -70°C refrigerator for later use.

Embodiment 3

[0041] Example 3: Construction of injection-type tissue engineered bone.

[0042] Injectable tissue engineered bone consists of solution A and solution B. Solution A is to dissolve fibrinogen with aprotinin solution and mix it with PRP at a ratio of 3 to 6:1, and add bone marrow stromal stem cells induced to differentiate and proliferate in vitro according to the method described in Example 1, and the cell density is 5× 10 6 -1×10 8 / ml. The concentration of aprotinin is 2000~6000U / ml. Solution B is calcium chloride and thrombin solution, the concentration of calcium chloride is 40mmol / ml, and the concentration of thrombin is 400U / ml. When in use, draw solution A and solution B separately with a double syringe. Mix and inject solutions A and B at a ratio of 9:1 to 1:1. It forms a jelly-like gel within 20-30 seconds when injected into the body, and also forms a gel in vitro.

[0043] Such as figure 1 As shown in the scanning electron microscope, the gel is a fibrous gri...

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Abstract

The present invention discloses one kind of injected tissue engineering bone and its construction and application. The injected tissue engineering bone consists of solution I and solution II, the solution I is the mixture of platelet rich plasma (PRP) and 80-100 mg / ml concentration fibrinogen solution in the ratio of 1 to 3-6, and contains marrow matrix stem cell in the concentration of 5*10<>6< / >-10*10<>7< / > / ml; and the solution II is solution mixture of calcium chloride in 40 mmol / ml concentration and thrombin in 400-1000 U / ml concentration. When the injected tissue engineering bone is used, solution I and solution II in the volume ratio of 1-9 are sucked with two syringes and injected into body to form gel with high adhesive force, with PRP accounting for 10-30 vol%. The injected tissue engineering bone is used in preparing implant for treating bone defect and bone nonunion and promoting fracture healing.

Description

technical field [0001] The invention belongs to the technical field of constructing artificial organs by tissue engineering method in biomedical engineering, and in particular relates to an injection type tissue engineering bone and its construction and application. Background technique [0002] At present, tissue engineered bone is mostly constructed as a model in which the scaffold material is loaded with cell growth factors and then compounded with seed cells. The selection of scaffold materials and growth factors is the focus of tissue engineering research. Scaffold materials, as seed cell carriers, have the functions of loading cells and slowing the release of growth factors. At present, most commonly used scaffold materials are solid materials, which have achieved remarkable results in the construction of tissue engineered bone. However, there are also defects such as difficulty in shaping, complex operation of loading growth factors and seed cells, low loading rate, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/19A61K47/42A61L27/22A61P19/08
Inventor 裴国献黄爱文金丹曾宪利胡稷杰陈书军张元平
Owner NAN FANG HOSPITAL
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