Evaluation of the Osteogenic Induction of Adipose Tissue-Derived Mesenchymal Stem Cells by OCS-B Collagen Scaffold and Osteoblast-Derived Microvesicles

Backgrounds: Tissue engineering is an interdisciplinary field that applies principals of life sciences and engineering to improve or replace failing or malfunctioning body tissues by combining cells, scaffolds and bioactive molecules. Scaffolds provide suitable matrix for cellular interaction and new functional tissues formation. Herein we investigated the osteogenic induction of adipose tissue-derived mesenchymal stem cells by OCS-B collagen scaffold and osteoblast-derived microvesicles.Materials and methods: Adipose tissue-derived stem cells were isolated from 6-7 gram of rat adipose. After 7days treatment by osteogenic medium, the microvesicles were isolated from the culture medium of the treated cells by ultracentrifugation. In the control group, no hormonal treatment was performed. The OCS-B scaffold was coated with 500,000 stem cells and treated with the microvesicles derived from untreated and treated stem cells separately. After 1, 3, and 5 days of incubation, alteration in osteogenic gene markers (RUNX2 and Osteonectin) were investigated by real-time PCR.Findings: data of SEM and DLS analyses confirmed the microvesicle isolation. Realtime PCR tests showed that both genes were upregulated significantly in microvesicle-treated group. Also, upregulations were higher on day 3 than day 5 .Conclusion: Altogether, the functional synergism between OCS-B scaffold and osteoblast-derived microvesicles for osteogenic differentiation of stem cells is reported. This complex would be a good candidate for future in vivo evaluations in order to be used in clinical applications.

Keywords:

Stem cells , extra membraneous micro vesicle , OCS-B scaffold