Effect of calcitonin gene-related peptide on osteoporotic fracture healing in vitro
10.3760/cma.j.issn.0253-2352.2017.06.005
- VernacularTitle:降钙素基因相关肽对骨质疏松性骨折愈合影响的体外研究
- Author:
Wei LIANG
;
Tiancheng LYU
;
Li LI
;
Bing LI
- Keywords:
Calcitonin gene-related peptide;
Osteoporosis;
Fractures,bone
- From:
Chinese Journal of Orthopaedics
2017;37(6):353-359
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of calcitonin gene-related peptide (CGRP) on proliferation of vascular endothelial cells (VECs) and osteogenic differentiation of bone mesenchymal stem cells (BMSCs) of osteoporotic rats in vitro,and cell/ molecular mechanism of promoting osteoporotic fracture healing.Methods 10 3-month-old SD rats were employed to make osteoporosis model by taking bilateral ovariectomy.Micro-CT analysis was used to determine the success of modeling at 6 months of postoperation.BMSCs and VECs were isolated from osteoporotic rats by blood adherence mcthod.10-7-10-10 mol/L CGRP were added to the culture medium.CCK-8 method was used to examine the cell proliferation.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR.Resuits BMSCs and VECs of osteoporotic rats were isolated successfully.The optical density (OD) values of CCK-8 method increased in a dose-dependent manner,for BMSCs at 7 d,the control group of OD value was 0.58±0.02,the CGRP group 10-7-10-10 mo]/L was to 0.80±0.03,0.79±0.03,0.74±0.03,0.69±0.03,difference was significant;for VECs,the control group of OD value was 2.20±0.01,the CGRP group 10-7-10-10 mol/L was to 4.12±0.04,3.90±0.02,3.56±0.04,3.12±0.04,difference was significant.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR,difference between the control group and CGRP group were all significant.Conclusion 10-7 mol/L,10-10 mol/L of CGRP can directly promote the proliferation of the BMSCs and VECs of osteoporotic rats,promote the osteogenic differentiation of BMSCs,improve the expression of VEGF gene and protein secretion,promote angiogenesis.
