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

具有亲水/疏水微酶分离结构的聚（2-羟乙基甲基丙烯酸酯）-聚苯乙烯(PSt)-PHEMA aba型嵌段共聚物（HSB）表面可抑制淋巴细胞坏死。而PSt和PHEMA-PSt无规则共聚物（HSR）表面引起淋巴细胞坏死。为了阐明其抑制机制：1)透射电镜（TEM）分析了粘附于HSB表面并与之接触的淋巴细胞的糖萼；2)分别用扫描电镜（SEM）和透射电镜（TEM）分析经神经氨酸酶处理的淋巴细胞与HSB表面的粘附和接触情况。采用图像处理器/分析仪（IA）对接触淋巴细胞和线粒体的TEM图像进行定量评价；3)用扫描电镜（SEM）和透射电镜（TEM）分析二甲基亚砜（DMSO）处理后的淋巴细胞粘附在HSB表面的情况。用IA定量评价黏附淋巴细胞和线粒体的TEM图像。用乳香红对淋巴细胞膜糖萼进行染色。用神经氨酸酶将唾液酸从淋巴细胞质膜的糖蛋白中去除。用DMSO抑制淋巴细胞细胞膜表面免疫球蛋白的形成。PSt和HSR表面作为对照聚合物。用微球柱法研究了聚合物表面与淋巴细胞的相互作用。整个g；与HSB表面黏附和接触的淋巴细胞的糖萼与完整淋巴细胞相同。神经胺酶处理和dmso处理的淋巴细胞，黏附和接触聚合物表面的淋巴细胞均呈圆形，细胞质保持良好。计算机TEM图像分析并没有显示所有病例的显著差异。提示HSB的微相分离结构表面保持了质膜结构和功能的稳定，从而抑制了淋巴细胞的坏死。少

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