The proliferation and tenogenic differentiation potential of bone marrow-derived mesenchymal stromal cell are influenced by specific uniaxial cyclic tensile loading conditions

Abstract

It has been previously demonstrated that mechanical stimuli are important for multipotent human bone marrow-derived mesenchymal stromal cells (hMSCs) to maintain good tissue homeostasis and even to enhance tissue repair processes. In tendons, this is achieved by promoting the cellular proliferation and tenogenic expression/differentiation. The present study was conducted to determine the optimal loading conditions needed to achieve the best proliferation rates and tenogenic differentiation potential. The effects of mechanical uniaxial stretching using different rates and strains were performed on hMSCs cultured in vitro. hMSCs were subjected to cyclical uniaxial stretching of 4, 8 or 12 % strain at 0.5 or 1 Hz for 6, 24, 48 or 72 h. Cell proliferation was analyzed using alamarBlue[Formula: see text] assay, while hMSCs differentiation was analyzed using total collagen assay and specific tenogenic gene expression markers (type I collagen, type III collagen, decorin, tenascin-C, scleraxis and tenomodulin). Our results demonstrate that the highest cell proliferation is observed when 4 % strain [Formula: see text] 1 Hz was applied. However, at 8 % strain [Formula: see text] 1 Hz loading, collagen production and the tenogenic gene expression were highest. Increasing strain or rates thereafter did not demonstrate any significant increase in both cell proliferation and tenogenic differentiation. In conclusion, our results suggest that 4 % [Formula: see text] 1 Hz cyclic uniaxial loading increases cell proliferation, but higher strains are required for superior tenogenic expressions. This study suggests that selected loading regimes will stimulate tenogenesis of hMSCs.

Fate of tenogenic differentiation potential of human bone marrow stromal cells by uniaxial stretching affected by stretch-activated calcium channel agonist gadolinium

“…It is worth noting that whilst calcium signalling may control many facets of cell behaviour, in this study we have limited our investigation to only cell differentiation and cell death; since it is our intention to establish the rate and strain levels that would provide the optimal cell response. This is in line with the study we conducted previously [19]. …”

“…Human bone marrow MSCs isolation was performed using our standard laboratory protocol, also previously described in other publications [19, 22]. Briefly, diluted bone marrow specimens were slowly layered on top of Ficoll–Paque PREMIUM (density of 1.077 g/mL) (Amersham Biosciences, Uppsala, Sweden).…”

“…The cells were serially passaged and expanded until passage-2 before being used for our experiments. The hMSCs surface markers analysis and tri-lineage differentiation was performed following standards established as our standard laboratory protocol, which was also mentioned in our previous publications [19]. …”

Strategies to engineer tendon/ligament-to-bone interface: Biomaterials, cells and growth factors

“…Although most studies have focused only on one stress component so far, the results obtained have provided key insights into the influence of mechanical loading on MSC differentiation. A number of studies report that MSCs subjected to cyclic tensile strain upregulate the expression of tenogenic genes such as Sxc, collagen type I and collagen type III [167][168][169][170][171][172]. In addition, cyclic tensile strain has also been shown to stimulate osteogenesis of MSC, upregulate the expression of the osteogenic genes BMP-2 and alkaline phosphatase, and increase mineral matrix deposition [38,[173][174][175][176][177][178][179].…”

Use of teriparatide off-label: our experience and review of literature

“…On the other side, an optimal balance between stability and micro-motion encourages callus formation and produces electrical local currents which improve bone healing by stimulating an inflammatory-type response and promoting calcification of fibrocartilage. Low-intensity pulsed ultrasound plays a relevant role in biomechanical environment, it accelerates fracture healing perhaps by acting directly to proliferation and differentiation of multipotent human bone marrow-derived mesenchymal stromal cells (2). Actually, the use of Platelet-Rich Plasma (PRP) seems to play an important role in the fracture healing and to favor fusion processes in spinal arthrodesis (3).…”