Abstract
Vascular tissue engineering approaches often require autologous smooth muscle cells (SMCs) obtained by biochemical differentiation of donor-derived adipogenic mesenchymal stem cells (ASCs) by transforming growth factor-β (TGFβ), bone morphogenetic protein-4 (BMP4), and/or sphingosylphosphorylcholine (SPC). However, data on myogenic marker proteins and contractility indicating fully functional SMC are contradicting and incomplete. We here investigated (1) the ideal combination of TGFβ, BMP4, and/or SPC; the time frame (2) necessary to trigger myogenic differentiation and (3) for a sustained differentiation status up to 29 days; and (4) donor diversity in myogenic differentiation potential and contractility using collagen and fibrin gel assays. Western blots showed that a combination of TGFβ, BMP4, and SPC most effectively induced both expression of α-smooth muscle actin (αSMA) and smoothelin after 8 days and their maintenance up to 29 days. Moreover, only three of seven ASC donors revealed effective coexpression of αSMA, smoothelin, and myosin heavy chain 11 and pronounced SMC contractility, whereas the latter was weaker (61%) and 6 days delayed in fibrin gels. Thus, a thorough biochemical and functional characterization of differentiated SMC is necessary before their use for autologous vascular graft cellularization. Moreover, strategies to improve fibrin scaffolds in terms of contractility should be developed.
Impact Statement
We here showed that even under optimized conditions for biochemical differentiation of adipose-derived stem cells (with respect to a pronounced marker protein expression for a reasonable period of time) it was not possible to obtain functional smooth muscle cells from all donors. Moreover, an underestimated role may play the effect of the scaffold material on smooth muscle cell functionality. Both aspects are crucial for the successful tissue engineering of the vascular medial layer combining autologous cells with a suitable scaffold material and thus should be thoroughly addressed in each individualized therapeutic approach.
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