Effect of mesenchymal stem cells and their niche on diabetic and osteoporotic wound healing following osteogenic differentiation and bone matrix formation in vitro

Yıl 2019, Cilt: 13 Sayı: 3, 183 - 192, 31.12.2019

Müge Karakayalı Tuna Önal , Mehmet İbrahim Tuğlu

Öz

Objectives: Mesenchymal stem cells (MSC) and their secreted factors (i.e. niche) are becoming growingly popular in bone
regeneration. The mechanisms of this effect can be investigated through in vitro models which are cost-effective methods
used for determining the effectiveness of new products in experimental and clinical applications. In the present study, we
established an experimental diabetic osteoporosis model in a high-glucose culture medium with no estrogen supplement to
investigate 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 to
assess osteoblastic differentiation and bone islet formation. A wound model was induced using the scratch assay to investigate
the 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 and
this 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 treatment
model in wound healing in patients with diabetic osteoporosis. Moreover, this model could be a cost-effective method for
the new treatment products to be applied in dental and orthopedic practice prior to animal experiments and clinical trials.

Anahtar Kelimeler

diabetic osteoporosis, in vitro, mesenchymal stem cell, niche, osteoblastic differentiation

Kaynakça

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