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壳聚糖 - 聚乙烯吡咯烷酮水凝胶作为胰岛免疫隔离的候选材料:体外生物相容性评估

Chitosan-polyvinyl pyrrolidone hydrogels as candidate for islet immunoisolation: in vitro biocompatibility evaluation.

作者信息

Risbud M, Hardikar A, Bhonde R

机构信息

Tissue Engineering and Banking Laboratory, National Centre for Cell Science, Pune, India.

出版信息

Cell Transplant. 2000 Jan-Feb;9(1):25-31. doi: 10.1177/096368970000900104.

Abstract

The success of immunoisolation devices for islet transplantation depends on the nature of semipermeable membranes, which permit the crossover of micronutrients, glucose, and insulin and prevent the entry of immunocytes and other transplant rejection mechanisms. In the present study we examined the properties of chitosan-polyvinyl pyrrolidone (PVP) hydrogels for possible application as an immunoisolation device. Hydrogels with two different proportions of chitosan-PVP (M1 1:1 and M2 2:1, v/v) were synthesized by cross-linking with glutaraldehyde. Hydrogels were characterized for their hydrophilic nature, protein adsorption, diffusion properties, cytotoxicity, and islet compatibility. Hydrogel membranes were found to be hydrophilic as determined by high octane contact angle value (M1: 142.9 +/- 0.46; M2: 143.6 +/- 0.49). Protein adsorption on the hydrogels was found to be low (0.0143 +/- 0.0027 mg for M1 and 0.0136 +/- 0.0049 mg for M2) compared to tissue culture polystyrene (TCPS) (0.0434 +/- 0.001 mg) and pure chitosan (0.0214 +/- 0.0025 mg) control. Hydrogel M1 was tested as a representative for diffusion studies. M1 allowed regulated transport of insulin and did not allow anti-insulin antibodies to pass through. In vitro biocompatibility of M1 and M2 was found to be excellent with no cytotoxic effects on the HeLa cells as determined by MTT and NR assay. Mouse islets cultured on the hydrogel membranes retained their integrity and intact morphology as assessed by image analysis study. Viability of islets cultured on hydrogels was comparable to that of controls (M1: 97%; M2: 90.4%) as assessed by trypan blue dye exclusion test. Islets retained their functionality when cultured on hydrogels, as judged by insulin secretion in response to glucose challenge (16.0 mM). Although in vivo experiments are awaited, the present study provides sufficient documentation to consider chitosan-PVP membranes as potential candidates for immunoisolation of islets.

摘要

用于胰岛移植的免疫隔离装置的成功取决于半透膜的性质,这种半透膜允许微量营养素、葡萄糖和胰岛素穿过,并防止免疫细胞和其他移植排斥机制的进入。在本研究中,我们研究了壳聚糖 - 聚乙烯吡咯烷酮(PVP)水凝胶作为免疫隔离装置的可能应用特性。通过与戊二醛交联合成了两种不同比例壳聚糖 - PVP(M1 1:1和M2 2:1,v/v)的水凝胶。对水凝胶的亲水性、蛋白质吸附、扩散特性、细胞毒性和胰岛相容性进行了表征。通过高辛烷接触角值测定发现水凝胶膜具有亲水性(M1:142.9±0.46;M2:143.6±0.49)。与组织培养聚苯乙烯(TCPS)(0.0434±0.001 mg)和纯壳聚糖(0.0214±0.0025 mg)对照相比,发现水凝胶上的蛋白质吸附较低(M1为0.0143±0.0027 mg,M2为0.0136±0.0049 mg)。水凝胶M1作为扩散研究的代表进行了测试。M1允许胰岛素的调节转运,不允许抗胰岛素抗体通过。通过MTT和NR测定法确定,M1和M2的体外生物相容性极佳,对HeLa细胞无细胞毒性作用。通过图像分析研究评估,在水凝胶膜上培养的小鼠胰岛保持其完整性和完整形态。通过台盼蓝染料排除试验评估,在水凝胶上培养的胰岛活力与对照相当(M1:97%;M2:90.4%)。当在水凝胶上培养时,胰岛在葡萄糖刺激(16.0 mM)下的胰岛素分泌表明其保留了功能。尽管尚待进行体内实验,但本研究提供了充分的证据,可将壳聚糖 - PVP膜视为胰岛免疫隔离的潜在候选材料。

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