葛根素参与的可注射钙离子敏感型水凝胶生物相容性研究

Biocompatibility evaluation of a puerarin-interact,injectable and Ca2+ responsive hydrogel

关键词:  葛根素  海藻酸钙  离子敏感型水凝胶  生物相容性

Key words:  puerarin  calcium alginate  ion responsive hydrogel  biocompatibility

摘要:

[目的] 将用于关节炎治疗的钙离子敏感型葛根素-海藻酸钙可注射水凝胶进行体内生物相容性评价研究。[方法] 制备不同处方配比的葛根素-海藻酸钙水凝胶，通过大鼠皮下注射埋植实验，考察葛根素-海藻酸钙水凝胶的体内降解情况；同时通过实时定量逆转录-聚合酶链反应（RT-PCR）和酶联免疫吸附（ELISA）分别测定注射水凝胶周围组织炎性相关因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的基因表达和蛋白分泌变化；通过组织切片苏木精-伊红（HE）染色观察不同埋植时间点水凝胶周围组织病理变化。[结果] 不同观察时间点各实验组凝胶直径无下降趋势，凝胶体内降解速率缓慢；与假手术组比较，葛根素-海藻酸钙凝胶周围组织中炎性因子IL-1β、IL-6和TNF-α蛋白浓度显著降低，而IL-1β、IL-6和TNF-α的mRNA表达无显著变化；组织切片HE染色结果表明在皮下结缔组织层产生短期物理损伤炎症反应，长期埋植后葛根素-海藻酸钙凝胶周围组织无炎症反应，具有良好的生物相容性。[结论] 研究中构建的可注射葛根素-海藻酸钙凝胶，体内降解缓慢，生物相容性良好，有望成为治疗骨关节炎的良好药物及细胞载体。

Abstract:

[Objective] To evaluate the biocompatibility of calcium ion sensitive injectable hydrogel with puerarin for arthritis treatment in vivo.[Methods] The degradation of puerarin-calcium alginate hydrogel was investigated in rats,which was implanted with different formulations of puerarin-calcium alginate hydrogel. The protein expression of inflammatory cytokines such as IL-1β,IL-6,and TNF-α were determined by ELISA,and the mRNA expressions of IL-1β,IL-6 and TNF-α were analyzed by real time RT-PCR in the tissue around the hydrogel. HE staining was used to observe the pathological changes of tissues around the hydrogel at different times.[Results] There was no significant decrease of the hydrogel volume in different time,which indicated that the rate of the hydrogel degradation was very slow in vivo. Compared with the sham-operated group,the protein levels of IL-1 β,IL-6 and TNF-α were significantly decreased in puerarin-calcium alginate gel group,but the mRNA expression of IL-1β,IL-6 and TNF-α was not significantly changed. HE staining results showed that puerarin-calcium alginate hydrogel only produced short-term inflammatory response by physical damage in the subcutaneous connective tissue layer,but no inflammatory response after long-term implantation,which indicated that the hydrogel had good biocompatibility.[Conclusion] In this study,injectable puerarin-calcium alginate hydrogel with suitable strength was constructed,which has the characteristics of slow degradation and good biocompatibility in vivo. It is expected to be excellent drug and cell carriers for the treatment of osteoarthritis.