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Efficient repairing effect of PEG based tri-block copolymer on mechanically damaged PC12 cells and isolated spinal cord
Authors:Iman Rad  Hamid Mobasheri  Farhood Najafi  Maryam Rezaei
Affiliation:1. Laboratory of Membrane Biophysics and Macromolecules, Institute of Biochemistry & Biophysics, University of Tehran, PO Box 13145-1384, Tehran, Iran
2. Biomaterials Research Centre (BRC), University of Tehran, PO Box 13145-1384, Tehran, Iran
3. Department of Resin and Additives, Institute for Color Science and Technology, Tehran, Iran
Abstract:Membrane sealing effects of polymersomes made of tri-block copolymer, PEG-co-FA/SC-co-PEG, (PFSP) were studied on isolated spinal cord strips, PC12 cell lines and artificial bilayer following mechanical impact implemented by aneurism clip, sonication and electric shock, respectively. The homogeneity and size of PFSP, membrane permeability and cell viability were assessed by dynamic light scattering, LDH release and MTT assays. According to the results, the biocompatible, physico-chemical, size, surface charge and amphipathic nature of PFSP polymersome makes it an ideal macromolecule to rapidly reseal damaged membranes of cells in injured spinal cord as well as in culture medium. Compound action potentials recorded from intentionally damaged spinal cord strips incubated with PFSP showed restoration of neural excitability by 82.24 % and conduction velocity by 96.72 % after 5 min that monitored in real time. Thus, they triggered efficient instant and sustained sealing of membrane and reactivation of temporarily inactivated axons. Treatment of ultrasonically damaged PC12 cells by PFSP caused efficient cell membrane repair and led to their increased viability. The optimum effects of PFSP on stabilization and impermeabilizing of the lipid bilayer occurred at the same concentrations applied to the damaged cells and spinal cord fibers and was approved by restoration of membrane conductance and calcein release manifested by NanoDrop technique. The unique physico-chemical characteristics of novel polymersomes introduced here, make them capable to reorganize membrane lipid molecules, reseal the breaches and restore the hydrophobic insulation in spinal cord damaged cells. Thus, they might be considered in the clinical treatment of SCI at early stages.
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