Effect of mesenchymal stem cells and their niche on diabetic and osteoporotic wound healing following osteogenic differentiation and bone matrix formation in vitro
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2019
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Abstract
Objectives: Mesenchymal stem cells (MSC) and their secreted factors (i.e. niche) are becoming growingly popular in boneregeneration. The mechanisms of this effect can be investigated through in vitro models which are cost-effective methodsused for determining the effectiveness of new products in experimental and clinical applications. In the present study, weestablished an experimental diabetic osteoporosis model in a high-glucose culture medium with no estrogen supplement toinvestigate the effect of MSC and their niche which their factors secreted into 24 hours medium on osteoblastic differentiation, formation of bone islets, and the wound healing model induced by scratch assay.Methods: A culture medium of adipose-derived rat MSC (ADMSC) with no estrogen supplement was used for cell growth toassess osteoblastic differentiation and bone islet formation. A wound model was induced using the scratch assay to investigatethe effect of the model on the parameters of wound healing. Cell growth and viability was assessed using MTT assay, cell migration and differentiation and the amount of wound closure were assessed based on the expression of CD44, CD45, and CD73,and osteoblast differentiation was evaluated using Alizarin Red S and von Kossa staining. Morphological observations were performed using an inverted phase-contrast microscope and h-score was assessed with immunohistochemical staining.Results: The use of osteogenic medium with estrogen supplement led to MSC growth and migration as well as bone islet formation. The use of a high-glucose medium without estrogen supplement inhibited MSC differentiation and bone islet formation. The administration of MSC and niche promoted the wound healing initiated by the administration of the scratch assay andthis promotion was significant in terms of all the parameters of wound healing.Conclusion: The results indicated that the therapeutic effect of MSC and niche could be used as an effective treatmentmodel in wound healing in patients with diabetic osteoporosis. Moreover, this model could be a cost-effective method forthe new treatment products to be applied in dental and orthopedic practice prior to animal experiments and clinical trials.