Poloxamer 188 enhances functional recovery of lethally heat-shocked fibroblasts
F A Merchant1, W H Holmes, M Capelli-Schellpfeffer
1Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Boston, USA.
Abstract:
Damage to the cell membrane has been implicated as the primary event in the pathogenesis of heat shock, generally resulting in loss of cellular homeostasis and cell death. Thus a promising mode of therapy would involve the restoration of cell membrane integrity. Surfactant molecules, specifically triblock polymers such as Poloxamer 188 (P-188), possess the ability to self-aggregate into membrane-like structures in aqueous solutions and have been shown to restore membrane integrity. The objective of this study was to develop functional and morphological assays to determine whether treatment with P-188 after heat shock enhances the recovery of thermally damaged cells. Human foreskin fibroblasts were placed in sterile vials and heated by immersion in a calibrated water bath for various lengths of time at predefined temperatures. Cell recovery after heat shock was assessed using a functional assay based on the ability of the cells to contract fibroblast populated collagen lattices (FPCLs). Subsequent to heating, collagen lattices were prepared with control (no heat, no P-188) and heat shocked cells (with and without P-188). Our results indicate that treatment with low concentrations of P-188 after heat shock was effective in ameliorating both the morphological integrity and the contractile function of thermally damaged cells. Further, we observed that P-188 was most effective in improving the contractile ability of cells heat shocked at 45 degrees C; however, it had no influence on the contractility of cells exposed to higher temperatures. Our results suggest that there exists a threshold of thermal stress (45 degrees C for 20-60 min) beyond which treatment with low concentrations of P-188 (0.5 mg/ml) is ineffective in minimizing cell damage. Moreover, the results of our morphological assays indicate that cells treated with P-188 after heat shock maintain their cytoskeletal organization, whereas untreated cells exhibit filamentous actin depolymerization.
