Chitosan gel as an in situ-forming scaffold for rat bone marrow mesenchymal stem cells in vivo

We herein formulated and characterized an in situ-forming chitosan gel consisting of chitosan and glycerol phosphate (GP) disodium salt, and examined its use as an in vivo scaffold for rat bone marrow mesenchymal stem cells (rBMSCs). First, the phase transition behaviors of chitosan solutions formulated with and without GP were characterized as a function of temperature. Chitosan solutions containing > 20 wt% GP became a gel at 37 degrees C and maintained this form for 28 days in vitro and in vivo. Next, we examined whether the chitosan gel could act as a suitable biocompatible substrate for the attachment and proliferation of rBMSCs. Immunohistochemistry clearly demonstrated that rBMSCs survived well on the scaffold created by in situ-forming chitosan gel in rats. Injection of chitosan gel alone induced macrophage accumulation in the host tissue and at the edge of the chitosan, whereas injection of chitosan gel containing rBMSCs was associated with decreased macrophage accumulation, indicating immunosuppression by the transplanted rBMSCs. Our results collectively show for the first time that chitosan gel could serve as an in situ-forming gel scaffold for entrapped rBMSCs in vivo.