Thermosensitive polymers: synthesis, characterization, and delivery of proteins

Three triblock copolymers based on the poly(lactide) or poly(lactide-co-glycolide) and poly(ethylene glycol) or poly(ethylene oxide) blocks were synthesized and characterized. The weight average molecular weight and number average molecular weight were determined by gel permeation chromatography and proton nuclear magnetic resonance spectroscopy, respectively. Fourier transform infrared spectroscopy was used to determine the completion of synthesis of polymers. Thermoreversible sol-gel transition temperature and concentration were determined by an inverted tube method. Two formulations each of three synthesized polymers containing 5% (w/v) of lysozyme or bromelain but differing in polymer concentrations (20-30%, w/v) were prepared and studied for in vitro release of the incorporated protein. In vitro biocompatibility of the delivery systems was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability assay. Biological activities of lysozyme and bromelain were determined by enzyme activity assays. Critical gelling concentrations were found in the range of 20-30% (w/v). In vitro biocompatibility study showed that all the formulations were biocompatible. Increasing the polymer concentration led to a decrease in burst release and extended the in vitro release of proteins. Furthermore, biological activities of lysozyme and bromelain in released samples were found to be significantly (p<0.05) greater in comparison to the control. Thus, the above thermosensitive polymers were able to deliver proteins in biologically active forms at a controlled rate for 2-8 weeks.