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Abstract
Mesenchymal stem cells (MSCs) were obtained from human bone marrow and amplified in cultures supplemented with human platelet lysate in order to generate myofibroblasts. When MSCs were seeded in solid collagen scaffolds, they differentiated into myofibroblasts that were observed to strongly bind to the substrate, forming a 3D cell scaffold network that developed tension and shortening after KCl stimulation. Moreover, MSC-laden scaffolds recapitulated the Frank-Starling mechanism so that active tension increased in response to increases in the initial length of the contractile system. This constituted a bioengineering tissue that exhibited the contractile properties observed in both striated and smooth muscles. By using the A. F. Huxley formalism, we determined the myosin crossbridge (CB) kinetics of attachment (f1) and detachment (g1 and g2), maximum myosin ATPase activity, molar myosin concentration, unitary CB force and maximum CB efficiency. CB kinetics were dramatically slow, characterizing the non-muscle myosin type IIA (NMMIIA) present in myofibroblasts. When MSCs were seeded in solid collagen scaffolds functionalized with Arg-Gly-Asp (RGD), contractility increased and CB kinetics were modified, whereas the unitary NMMIIA-CB force and maximum CB efficiency did not change. In conclusion, we provided a non-muscle bioengineering tissue whose molecular mechanical characteristics of NMMIIA were very close to those of a non-muscle contractile tissue such as the human placenta.
Highlights
We reported that mesenchymal stem cells (MSCs) derived from human bone marrow (BM) and seeded in solid 3D-collagen scaffolds differentiated into myofibroblasts in the presence of human platelet lysate (HPL) and contracted when exposed to KCl or an electrical field [1]
In the subsections 1 and 2, physical properties of collagen scaffolds seeded with MSCs, with or without RDG, and their passive mechanical properties (Fig 2) were presented
The fundamental mechanical properties observed in all striated and smooth muscles were observed in MSC-laden scaffolds, namely the FrankStarling phenomenon and the A.V
Summary
We reported that mesenchymal stem cells (MSCs) derived from human bone marrow (BM) and seeded in solid 3D-collagen scaffolds differentiated into myofibroblasts in the presence of human platelet lysate (HPL) and contracted when exposed to KCl or an electrical field [1]. Myofibroblasts are non-muscle contractile cells [2] that encourage wound healing by secreting collagen and inducing wound contraction. In 2D culture, the differentiation of fibroblasts into myofibroblasts is encouraged by several factors such as the transforming growth factor- β (TGF-β) [9, 10]. This is present in high concentrations in HPL routinely used to amplify MSCs [11]. The presence of collagen and fibronectin and a certain degree of stiffness of the environment where cells reside, impact the differentiation of MSCs towards myofibroblasts [12, 13]
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