Risbud M, Bhonde M, Bhonde R
Tissue Engineering and Banking Laboratory, National Centre for Cell Science, Ganeshkhind, Pune, India.
Cell Transplant. 2001 Mar-Apr;10(2):195-202.
We have shown previously that chitosan-polyvinyl pyrrolidone (PVP) hydrogels are blood compatible, islet compatible, and noncytotoxic to various cell types. Because of these potential applications of chitosan-PVP hydrogel, the present study was designed to investigate its effect on macrophage activation. Macrophages did not adhere to hydrogel in culture but maintained their viability and did not undergo apoptosis as confirmed by trypan blue staining and absence of DNA ladder. Hydrogel leach-out products did not exhibit cytotoxic effects on macrophage functionality at mitochondrial and lysosomal level as confirmed by tetrazolium reduction (MTT) and neutral red uptake (NRU) assay. On exposure to hydrogels, macrophages showed comparable expression of activation markers such as CDllb/CD18 (Mac-1), CD45, and CD14 to those cultured in the presence of PTFE, a known biocompatible control, indicating its nonactivating nature. Macrophage activation was also assessed by checking the level of messenger RNA of inflammatory cytokines such as IL-6 and TNF-alpha by reverse transcriptase polymerase chain reaction (RT-PCR), which did not show stastistically significant difference (p > 0.05) in the expression of these transcripts in both control and hydrogel-exposed macrophages. The nonimmunogenic nature of the hydrogel was further confirmed by the lack of induced proliferation of mouse splenic lymphocytes after exposure to hydrogel leach-outs. All these results point out that chitosan-PVP hydrogel did not activate macrophages and thus is immunocompatible. Our results indicate that this hydrogel could be a potential candidate for transplantation studies by virtue of its biocompatibility and imunocompatibility.
我们之前已经表明,壳聚糖-聚乙烯吡咯烷酮(PVP)水凝胶具有血液相容性、胰岛相容性,并且对各种细胞类型无细胞毒性。由于壳聚糖-PVP水凝胶具有这些潜在应用,本研究旨在探讨其对巨噬细胞活化的影响。巨噬细胞在培养中不粘附于水凝胶,但保持其活力且未发生凋亡,这通过台盼蓝染色和无DNA梯带得以证实。如通过四唑盐还原(MTT)和中性红摄取(NRU)试验所证实,水凝胶浸出产物在线粒体和溶酶体水平上对巨噬细胞功能未表现出细胞毒性作用。在接触水凝胶后,巨噬细胞显示出与在已知生物相容性对照聚四氟乙烯(PTFE)存在下培养的巨噬细胞相当的活化标志物如CD11b/CD18(Mac-1)、CD45和CD14的表达,表明其非活化性质。还通过逆转录聚合酶链反应(RT-PCR)检测炎性细胞因子如IL-6和TNF-α的信使RNA水平来评估巨噬细胞活化,在对照和接触水凝胶的巨噬细胞中这些转录物的表达未显示出统计学上的显著差异(p>0.05)。水凝胶浸出物接触后小鼠脾淋巴细胞缺乏诱导增殖进一步证实了水凝胶的非免疫原性。所有这些结果表明,壳聚糖-PVP水凝胶未激活巨噬细胞,因此具有免疫相容性。我们的结果表明,由于其生物相容性和免疫相容性,这种水凝胶可能是移植研究的潜在候选物。