The effects of bone marrow stromal cells transferred with lentivirat—mediated Sonic Hedgehog (Shh) gene in osteoporosis

doi:10.3877/cma.j.issn.2096-0263.2018.06.005

Abstract

Abstract:

Objective

To observe osteogenous differentiation of modified bone marrow stromal cells (BMSCs) which can express Sonic Hedgehog (Shh) stably and provide the treatment of osteoporosis in vivo feasibility.

Methods

BMSCs were infected with lentiviruses using a lentiviral vector containing the DsRed or the Shh-DsRed gene and then divided into Shh-DsRed- BMSCs group (group A) and DsRed- BMSCs group (group B). The transfection efficiency were evaluated by fluorescence microscopy using DsRed expression. The expression of Shh was tested by RT-PCR and Western-blot analysis after 48 hours. The osteogenous differentiation of BMSCs were analysed by ALP testing after 7 days, and tested by alizarin red staining after 28 days.

Results

The transfeetion efficiency was about 90% after 24 hours in Shh-DsRed-BMSCs. Compared with the group B, the Shh-DsRed-BMSCs can stably secrete Shh mRNA and protein. The ALP activity in Shh-DsRed-BMSCs group was significantly greater than those in DsRed-BMSCs. The alizarin red staining expression in group A was markedly higher than those in group B.

Conclusion

Shh-DsRed-BMSCs can long-term and stably secrete high levels of Shh, has a high ability of osteoblast differentiation and was then the basis for the subsequent treatment for osteoporosis in vivo.

Key words: Mesenchymal stromal cells, Osteoporosis, Transfection, Sonic hedgehog

Yijia Jia, Jiping Sun, Qiang Liu, Haihu Hao, Jian Zhu, Dou Wu. The effects of bone marrow stromal cells transferred with lentivirat—mediated Sonic Hedgehog (Shh) gene in osteoporosis[J]. Chinese Journal of Geriatric Orthopaedics and Rehabilitation(Electronic Edition), 2018, 04(06): 341-345.

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Figures/Tables 6

References 16

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