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

聚甲基丙烯酸羟乙酯(PHEMA)-聚苯乙烯(PST)-PHEMA-ABA型嵌段共聚物(HSB)表面具有亲水/疏水微相分离结构，可抑制淋巴细胞的坏死。相反，PST和PHEMA-PST无规共聚物(HSR)表面可引起淋巴细胞坏死。为了阐明抑制作用的机制：1)用透射电子显微镜分析淋巴细胞与HSB表面的粘连和接触；2)神经氨酸酶处理的淋巴细胞分别用扫描电子显微镜和透射电子显微镜分析其与HSB表面的粘连和接触情况。用图像分析仪(IA)对接触的淋巴细胞和线粒体的透射电子显微镜图像进行定量分析；3)用扫描电子显微镜和透射电子显微镜分析二甲基亚砜(DMSO)处理的淋巴细胞在HSB表面的黏附情况。免疫荧光定量分析贴壁淋巴细胞和线粒体的透射电子显微镜图像。Rutheniu…用更多的米红对淋巴细胞膜的糖基化进行染色。神经氨酸酶用于去除淋巴细胞质膜糖蛋白中的唾液酸。用DMSO抑制淋巴细胞膜表面免疫球蛋白的形成。以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。与HSB表面贴附和接触的淋巴细胞与完整淋巴细胞的整个溶菌体是相同的。经Neuramindase处理和DMSO处理的淋巴细胞分析表明，与聚合物表面贴附和接触的淋巴细胞均呈圆形，保持良好的细胞质。计算机化的电子显微镜图像分析并未显示所有病例均有显著差异。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。较少

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