Method for detecting and evaluating in-vitro cell morphology and osteogenic function of surface of bone substitute implant

A technology for bone implant materials and cell morphology, which is applied in material inspection products, analytical materials, material excitation analysis, etc., can solve problems such as insufficient comprehensiveness, shallow evaluation, and large limitations, achieve accurate measurement methods, and improve detection standards. , the effect of high detection level

Inactive Publication Date: 2015-12-02
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

Existing methods for detecting cell morphology and osteogenic function in vitro of bone implant materials are single, poor in accuracy, shallow in evaluation, not comprehensive enough, and have great limitations

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  • Method for detecting and evaluating in-vitro cell morphology and osteogenic function of surface of bone substitute implant
  • Method for detecting and evaluating in-vitro cell morphology and osteogenic function of surface of bone substitute implant
  • Method for detecting and evaluating in-vitro cell morphology and osteogenic function of surface of bone substitute implant

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Embodiment Construction

[0051] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0052] 1. Main materials and instruments

[0053] Improved α-MEM medium (Hyclone, USA); fetal bovine serum (Sijiqing, Zhejiang Tianhang Biotechnology Co., Ltd.); trypsin (Sigma, USA); penicillin / streptomycin (Sigma, USA); Glutamine; CCK-8 Kit (Wuhan Boster Bioengineering Co., Ltd., China); DMSO (Sigma, USA); Alkaline Phosphatase Detection Kit (Shanghai Biyuntian Biotechnology Co., Ltd., China); 1% Sirius Red / Picric Acid Staining Reagent (Shanghai Beyotime Biotechnology Co., Ltd., China); Alizarin Red Reagent (Sigma, USA); 2.5% Glutaraldehyde; 75% Ethanol (Tianjin Fuyu Fine Chemical Co., Ltd.); Rhodamine Labeling - Phalloidin (Sigma, U.S.); E.Z.N.A.TMTotalRNAKitI total RNA extraction kit (Omega, U.S.); PrimeScriptTMRTreagentkit reverse transcription kit (Takara, China); SYBRPremixExTaqIITM fl...

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Abstract

The invention discloses a method for detecting and evaluating in-vitro cell morphology and osteogenic function of the surface of a bone substitute implant, and belongs to the technical field of biomaterial detection. The method is characterized by comprising the following steps: taking the bone substitute implant to be detected and evaluated, firstly, observing the cell morphology, cytoskeleton morphology and skeleton protein distribution condition, then detecting cell activity, cell adhesion on the surface of the material and alkaline phosphatase activity in cells, then detecting collagen secretion and the external parts of the cells, and remembering the mineralization degree, and finally, determining the cell morphology as well as the expression condition of framework related genes and osteogenesis related genes. According to the method, biological reference is better provided for optimal design of the surface of the bone substitute implant, and on the basis of the prior art, lots of detecting methods are added, and part detecting methods are improved, so that the method is comprehensive and systematical, and passes the cytology and molecule biology cross comprehensive evaluation, and important guiding significances are provided for optimal research of the surface of the bone substitute implant.

Description

technical field [0001] The invention belongs to the technical field of biomaterial detection, and in particular relates to a method for detecting and evaluating cell morphology and osteogenic function on the surface of a bone implant material in vitro. Background technique [0002] A large number of studies have shown that the surface morphology and surface chemical properties of bone implant materials have a decisive impact on the osseointegration ability of implant materials, and their surface physical and chemical characteristics are crucial to the impact of osseointegration. Therefore, more and more surface treatment technologies focus on changing the surface characteristics of bone implant materials, and strive to improve the efficiency of implant materials and bone integration. At present, the surface activation treatment methods of bone implant materials include chemical methods, physical methods, physicochemical methods, and biochemical methods. method, calcium phos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12Q1/06C12Q1/68C12Q1/42G01N33/68G01N21/84G01N21/31G01N21/64
Inventor 李广文张玉梅王薇宋艳艳常蓓宋文刘茜赵领洲闫钧张力马千里方科达丁鑫
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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