Healthy cells are required in large numbers to form a tissue-engineered construct and primary cells must therefore be increased in number in a process termed 'expansion'. There are significant problems with existing procedures, including cell injury and an associated loss of phenotype, but three-dimensional culture has been reported to offer a solution. Reversible gels, which allow for the recovery of cells after expansion would therefore have great value in the expansion of chondrocytes for tissue engineering applications, but they have received relatively little attention to date. In this study, we examined the synthesis and use of thermoresponsive polymers that form reversible three-dimensional gels for chondrocyte cell culture. A series of polymers comprising N-isopropylacrylamide (NIPAM) and styrene was synthesized before studying their thermoresponsive solution behaviour and gelation. A poly(NIPAM-co-styrene-graft-N-vinylpyrrolidone) variant was also synthesized in order to provide increased water content. Both random-and graft-copolymers formed particulate gels above the lower critical solution temperature and, on cooling, re-dissolved to allow enzyme-free cell recovery. Chondrocytes remained viable in all of these materials for 24 days, increased in number and produced collagen type II and glycosaminoglycans.

Highly interconnected and 3D porous bioactive hydroxyapatite (HAP) and Bioglass scaffolds have been fabricated by an adaptive version of camphene based foam reticulation (ARM) and camphene freeze casting (CFC) methods. Controlled sublimation of camphene during freeze casting at -78°C produced process optimized bioscaffolds with open, uniform, and interconnected porous structures. HAP and Bioglass scaffolds with desired porosity, pore size, and microtopography were successfully fabricated using polyurethane foam templates of appropriate structures. Macropores of 50-1100 μm with microporosity of 1-10 μm, known to facilitate cell adhesion and proliferation, were obtained. Compressive yield strength of 0.8 MPa close to the upper range of cancellous bone was achieved. The mean compressive strength of HAP scaffolds compared favorably with the theoretical model of porosity variation with strength and was higher than reported values. The nature of pore development, morphology, porosity, crystal structure, chemical composition, and thermal behavior were characterized using scanning electron and optical microscopy, X-ray diffraction, thermal analysis, and mercury porosimetry. These scaffolds are suited for nonstructural graft and were not cytotoxic in vitro when osteoblast-like MG63 cells were cultured with the HAP constructs. The cells attached indicated by cell metabolic activity by resazurin assay and spread well when cultured on the surface of the materials.

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Item Type Article Authors Swift, Thomas; Lapworth, J.; Swindells, K.; Swanson, L.; Rimmer, Stephen Citation Swift T, Lapworth J, Swindells K et al (2016) pH responsive highly branched poly(N-isopropylacrylamide) with trihistidine or acid chain ends. RSC Advances. 75: 71345-71350.