Determining in vitro osteogenic activites of bone substitute biomaterials
- VernacularTitle:生物骨替代材料成骨活性的体外测定
- Author:
Jinbu ZUO
;
Mingxue SUN
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2008;12(32):6385-6388
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Conventjonally,the osteoinduction of bone substitute materials is detected in vivo,which is unsatisfactory regarding the reliability,chronergy and precision,especially for a large amount of substitute materials.OBJECTIVE:To search an in vitro assay for determining in vitro osteogenic activities of bone graft substitutes.DESIGN:Controlled cytological trials.TIME AND SETTING:The experiments were carried out in the Institute of Orthopaedics.Chinese PLA General Hospital(Beijing,China)from August 2006 to May 2007.MATERIALS:C2C 12 cells was offered by the Cell Center of Peking Union Medical College;Human decalcified bone matrix and bone protein were offered by the Institute of Orthopaedics in Chinese PLA General Hospital;Type Ⅰ collagen extracted from bovine tendon was purchased from Beijing Yierkang Bioengineering Development Center;Recombinant human bone morphogenetic protein 2 was purchased from Hangzhou Gene Technology of Huadong Medicine Group.METHODS:By means of dialysis,a composite material was prepared with the bone protein extracted from human,human decalcified bone matrix and type Ⅰ collagen of bovine tendon.The samples of decalcified bone matrix.composite material and recombinant human bone morphogenetic protein 2 were respectively co-incubated with C2C12cells for 72 hours.Negative control group comprised pure cells without materials.Then C2C 1 2 cells were lysed and the lysate were assayed for the absorbance of alkaline phosphatase(ALP)and total protein by chromatometry.ALP is (the specific mark of osteoblastic phenotype.The relative ratio of absorbance value between ALP and total protein could represent ALP activity in the unit quantitative C2C12 cells.MAIN OUTCOME MEASURES:The ratio of absorbance value between ALP and total protein.RESULTS:The ALP activity was the highest in the recombinant human bone morphogenetic protein 2 group,then in the decalcified bone matrix group and composite material group,and the lowest in the negative control group.There were significant differences in the ALP activity between the three trial groups and the negative control group(P＜0.05).CONCLUSION:The assay in vitro is effective to detect the ALP activity and it can be used to determine the osteoinduction of bone graft substitutes.
