Bioreactor mechanically guided 3D mesenchymal stem cell chondrogenesis using a biocompatible novel thermo-reversible methylcellulose-based hydrogel.
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 Na2SO4solution. 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.
Laboratory of Biomedical Materials, Department of Health Sciences, Università del Piemonte Orientale "UPO", Novara, Italy.|Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), Firenze, Italy.
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Cochis
AO Research Institute Davos, Davos Platz, Switzerland.
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Grad
AO Research Institute Davos, Davos Platz, Switzerland.|University Medical Center, Albert-Ludwigs University Freiburg, Germany.
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Stoddart
Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), Firenze, Italy.|Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico di Milano, Milano, Italy.
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Farè
Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), Firenze, Italy.|Department of Chemistry, Materials and Chemical Engineering "G. Natta", Politecnico di Milano, Milano, Italy.
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Altomare
Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), Firenze, Italy.|Laboratory of Applied Microbiology, Department of Health Sciences, Università del Piemonte Orientale "UPO", Novara, Italy.
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Azzimonti
AO Research Institute Davos, Davos Platz, Switzerland.
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Alini
Laboratory of Biomedical Materials, Department of Health Sciences, Università del Piemonte Orientale "UPO", Novara, Italy.|Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), Firenze, Italy.
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Rimondini
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D015415:Biomarkers; D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D020219:Chondrogenesis / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D020100:Hydrogels; D008422:Materials Testing; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008747:Methylcellulose; D051379:Mice
D001672:Biocompatible Materials; D015415:Biomarkers; D020100:Hydrogels; D008747:Methylcellulose