J-C Xu1, G-H Wu, L-L Zhou, X-J Yang, J-T Liu. 1. Department of Orthopedics, Xiangyang Hospital, Hubei University of Medicine, Xiangyang, Hubei Province, China. guihua_wu1@163.com.
Abstract
OBJECTIVE: To study the effects of leptin (LEP) on the osteogenic differentiation of human bone marrow stromal cells (hBMSCs) and to explore the mechanism controlling the process. MATERIALS AND METHODS: Respectively cultivated the third-generation hBMSCs with 100 ng/ml bone morphogenetic protein (BMP) culture media containing 320, 160, 80 and 40 ng/mL LEP, and regular medium. We administered alkaline phosphatase (ALP) dye (on the 7th day) and mineralized nodules alizarin red (on the 21st day) and tested the ALP activity as well as osteocalcin (OCN) level on 7th, 14th, 21st day in each group to establish the best inducing concentration of LEP. 7 days later, we tested bone differentiation related genes expression in the control, 160 ng/mL LEP and 100 ng/mL BMP groups using RT-PCR. RESULTS: The activity of ALP and OCN in the 160 ng/mL LEP group after 7, 14 and 21 days was lower than that of the BMP group but higher than that of other groups. However, LEP significantly promoted the expression of bone differentiation related genes, namely, Cbfal, ALP, COL-I and OCN. CONCLUSIONS: LEP promoted the bone differentiation in hBMSCs by promoting the expression of genes related to bone differentiation.
OBJECTIVE: To study the effects of leptin (LEP) on the osteogenic differentiation of human bone marrow stromal cells (hBMSCs) and to explore the mechanism controlling the process. MATERIALS AND METHODS: Respectively cultivated the third-generation hBMSCs with 100 ng/ml bone morphogenetic protein (BMP) culture media containing 320, 160, 80 and 40 ng/mL LEP, and regular medium. We administered alkaline phosphatase (ALP) dye (on the 7th day) and mineralized nodules alizarin red (on the 21st day) and tested the ALP activity as well as osteocalcin (OCN) level on 7th, 14th, 21st day in each group to establish the best inducing concentration of LEP. 7 days later, we tested bone differentiation related genes expression in the control, 160 ng/mL LEP and 100 ng/mL BMP groups using RT-PCR. RESULTS: The activity of ALP and OCN in the 160 ng/mL LEP group after 7, 14 and 21 days was lower than that of the BMP group but higher than that of other groups. However, LEP significantly promoted the expression of bone differentiation related genes, namely, Cbfal, ALP, COL-I and OCN. CONCLUSIONS:LEP promoted the bone differentiation in hBMSCs by promoting the expression of genes related to bone differentiation.
Authors: Shu-Mei Chen; Yi-Jen Peng; Chih-Chien Wang; Sui-Lung Su; Donald M Salter; Herng-Sheng Lee Journal: Int J Med Sci Date: 2018-03-08 Impact factor: 3.738