ELUCIDATION OF THE INHIBITED MECHANISM OF IMMUNOCYTE DEATH ON THE SURFACE HYDROPHILIC/HYDROPHOBIC-TYPE MICROPHASE-SEPARATED STRUCTURE

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Poly (2-hydroxyethyl methacrylate) (PHEMA)-Polystyrene(PSt)-PHEMA ABA-type block copolymer (HSB) surfaces with hydrophilic/hydrophobic-type microhase-separated structure inhibited necrosis of lymphocytes. In contrast, PSt and PHEMA-PSt random copolymer (HSR) surfaces caused necrosis of lymphocytes. In order to clarify the inhibitory mechanism : 1) the glycocalyx of the lymphocytes adhered to and contacted with the HSB surfaces was analyzed by transmission electron microscopy(TEM) ; 2) Neuraminidase-treated lymphocytes adhered to, and contacted with the HSB surfaces were analyzed by scanning electron microscopy(SEM), and TEM respectively. The TEM images of the contacted lymphocytes and the mitochondria were evaluated quantitatively by an image processor / analyzer (IA) ; 3) Dimethyl sulfoxide (DMSO)-treated lymphocytes adhered to the HSB surfaces were analyzed by SEM and TEM. The TEM images of the adhered lymphocytes and the mitochondria were evaluated quantitatively by the IA. Rutheniu … More m red was used to stain the glycocalyx of the lymphocyte plasma membrane. Neuraminidase was used to remove the sialic acid from the glycoproteins of the lymphocyte plasma membrane. DMSO was used to inhibit the surface immunoglobulin capping formation of the lymphocyte plasma membrane. The PSt and HSR surfaces were used as control polymers. The interaction between the polymer surfaces and the lymphocytes was carried out by the microsphere column method. The entire g;lycocalyx of the lymphocytes adhered to and contacted with the HSB surfaces was the same as that of the intact lymphocytes. On analyses of Neuramindase-treated and DMSO-treated lymphocytes, the lymphocytes adhered to and contacted with the polymer surfaces were all observed to be round, keeping cytoplasm well. The computerized TEM image analysis did not indicate significant differences in all cases. It was suggested that the microphase-separated structure surfaces of the HSB inhibited the necrosis of the lymphocytes becasuse the surfaces kept the plasma membrane's structure and function stable. Less