Poloxamer 188 enhances functional recovery of lethally heat-shocked fibroblasts.
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.
Merchant
F A
FA
Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Boston, USA.
Holmes
W H
WH
Capelli-Schellpfeffer
M
M
Lee
R C
RC
Toner
M
M
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000473:pathology; D002460:Cell Line; D002470:Cell Survival; D003094:Collagen / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D018869:Heat-Shock Response / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D008297:Male; D011060:Poloxalene / Q000494:pharmacology*; D012879:Skin Physiological Phenomena / Q000187:drug effects*; D013501:Surface-Active Agents / Q000494:pharmacology*
D013501:Surface-Active Agents; D011060:Poloxalene; D003094:Collagen