Bioreactor mechanically guided 3D mesenchymal stem cell chondrogenesis using a biocompatible novel thermo-reversible methylcellulose-based hydrogel

Abstract

Autologous chondrocyte implantation for cartilage repair represents a challenge because strongly limited by chondrocytes’ poor expansion capacity in vitro. Mesenchymal stem cells (MSCs) can differentiate into chondrocytes, while mechanical loading has been proposed as alternative strategy to induce chondrogenesis excluding the use of exogenous factors. Moreover, MSC supporting material selection is fundamental to allow for an active interaction with cells. Here, we tested a novel thermo-reversible hydrogel composed of 8% w/v methylcellulose (MC) in a 0.05 M Na2SO4 solution. MC hydrogel was obtained by dispersion technique and its thermo-reversibility, mechanical properties, degradation and swelling were investigated, demonstrating a solution-gelation transition between 34 and 37 °C and a low bulk degradation (<20%) after 1 month. The lack of any hydrogel-derived immunoreaction was demonstrated in vivo by mice subcutaneous implantation. To induce in vitro chondrogenesis, MSCs were seeded into MC solution retained within a porous polyurethane (PU) matrix. PU-MC composites were subjected to a combination of compression and shear forces for 21 days in a custom made bioreactor. Mechanical stimulation led to a significant increase in chondrogenic gene expression, while histological analysis detected sulphated glycosaminoglycans and collagen II only in loaded specimens, confirming MC hydrogel suitability to support load induced MSCs chondrogenesis.

Three-dimensional cell culture of human mesenchymal stem cells in nanofibrillar cellulose hydrogels

“…To overcome these limitations, different 3D cultivation methods, more closely resembling the endogenous niche, have been applied to MSCs. Among others, MSCs can be cultivated in 3D within alginate hydrogels [8], collagen-based matrices [9], fibrin-poly(ester-urethane) scaffolds [10], bacteria-derived cellulose [11], methylcellulose [12] and the chondrosarcomaderived Matrigel TM [13]. However, despite the obvious advantages over the 2D cultivation hydrogels require cross-linking, while retrieval of the cells from fibrin and collagen based matrices can only be achieved by using enzymes also affecting mammalian cells.…”

New Insights on Mechanical Stimulation of Mesenchymal Stem Cells for Cartilage Regeneration

“…Cochis et al [35] cultivated MSCs in a methylcellulose solution retained within a porous polyurethane matrix to evaluate the suitability of the matrix in supporting mechanically induced chondrogenesis of the cells in the absence of exogenous factors. The composite underwent a combination of compression and shear forces by the use of a bioreactor which applied compressive and rolling movements through a ceramic ball.…”

“…Scaffolds embedded with ADSCs, subjected to cyclic compression in bioreactors, could benefit from the presence of chondrocytes, since they are able to release paracrine factors (i.e., TGF-β1 and IGF-1), guiding stem cells towards chondrogenic differentiation. This approach has been used to reduce the use of exogenous growth factors, which can instead be synthesized by a proper number of chondrocytes during in vitro expansion [35]. Even though similar production of type II collagen and GAGs was observed in single and co-cultures, the latter were shown to be able to suppress the expression of Col I, Col X, and Tumor necrosis factor alpha (TNF-α) markers [45].…”

New substrates for stem cell control

“…Fmoc-F 2 /S [42] pericyte chondrogenesis RADA-PRG (modified PuraMatrix w ) [43,44] mouse neonatal epidermal cells, rMSC neurogenesis RGD on decellularized pig heart valves [45] EPC proliferation Synthemax w [46] iPSC expansion and differentiation polymer enzymatically responsive PEG [47 -52] iPSC, hESC, mESC, MSC, organoids, mouse pancreatic progenitor gellan gum [53] hPSC 3D spheres methylcellulose [54] MSC chondrogenesis nanotopographically imprinted PCL [55,56] MSC maintenance and osteogenesis NO-releasing chitosan [57] hP-MSC angiogenic potential P(PEGMEMA-r-GMA-r-VDM) [58] MSC proliferation PEG with RGD and MMP tethering [59,60] MSC migration photodegradable PEG [61,62] MSC 'mechanical memory' polyacrylate/acrylamide thermoresponsive hydrogel [63 -65] hESC, MSC, mESC polyacrylate/polyurethane [66 -68] hESC, MSC, C6 rat GSC polypyrrole [69] MSC osteogenesis polystyrene TopoChip [70] iPSC proliferation polyurethane [71 -74] hESC-derived HE, HPC, iPSC-derived hepatocytes, H9 ternary polymer blends [75,76] STRO-1 þ skeletal SC, fetal skeletal SC rstb.royalsocietypublishing.org Phil. Trans.…”