Chondrogenic differentiation of human mesenchymal stem cells using a thermosensitive poly(N-isopropylacrylamide) and water-soluble chitosan copolymer

Poly(N-isopropylacrylamide) (PNIPAAm) is known to be thermally responsive material and has a lower critical solution temperature (LCST, 32°C) at which a macromolecular transition from a hydrophilic to a hydrophobic structure occurs. Chitosan is a useful natural polymeric biomaterial due to its biocompatibility and biodegradable properties. It has good characteristics for cell attachment, proliferation and viability. The aim of this study was to assess the ability to differentiate from mesenchymal stem cells (MSCs) to chondrocytes and mass formation using a newly developed injectable material, a thermosensitive (water-soluble chitosan-g-PNIPAAm) gel, and evaluate cartilage formation in vivo after injecting a cell-thermosensitive gel complex. The MSCs were cultured in the chitosan–PNIPAAm in vitro. Fluorescence-activated cell sort analysis, viability test, collagen type I, II, X formation and the aggrecan levels were examined. These cultured cells can be easily recovered from a copolymer gel by simply lowering the temperature. An animal study was performed to assess cartilage formation in the submucosal layer of the bladder of rabbits. The cartilage formation could be detected. This can be used to treat vesicoureteral reflux or reflux esophagitis by the effective mass effect. This is a simple method (sol–gel technique in LCST), and good cartilage formation occurs in the bladder tissue.