A novel method to in vitro evaluate biocompatibility of nanoscaled scaffolds
A novel method to in vitro evaluate biocompatibility of nanoscaled scaffolds
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体外评估纳米支架生物相容性的新方法
DOI：
10.1002/jbm.a.35743
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发表时间：
2016-09-01
影响因子：
4.9
通讯作者：
Watari, Fumio
中科院分区：
文献类型：
--
作者：
Li, Xiaoming;Wang, Zheng;Watari, Fumio
本研究为组织工程纳米支架与中性粒细胞的体外生物相容性评价提供了一种新的方法。从健康人外周血中分离出中性粒细胞。以纳米羟基磷灰石/胶原蛋白(NHAC)、纳米羟基磷灰石/胶原/聚L-乳酸(NHACP)和甲壳素纤维增强的nHACP(nHACP/CF)在D-Hank平衡盐溶液(D-HBSS)中的体外降解产物为测试液，然后将其与中性粒细胞混合。结果表明，试验液中的细胞存活率与D-HBSS(对照组)中的细胞存活率无显著差异。但从基因和蛋白表达水平来看，nHACP/CF测试液中中性粒细胞的乳酸脱氢酶和肿瘤坏死因子-α在整个测试过程中都是最低的，其主要原因可能是支架的钙释放速度最慢，其降解液的pH值最接近人体。体内实验表明，术后2周，nHAC组和聚乳酸L组小鼠背部皮下炎症反应最多，nHACP/CF组炎症反应最少，与体外实验结果一致。这些结果表明，纳米支架降解液的pH值和一定的金属铁浓度应该是显著影响其生物相容性的两个重要因素。本研究为生物可降解纳米组织工程支架材料提供了一种简单有效的生物相容性测试方法。(C)2016 Wiley期刊，Inc.生物材料资源A部分：104A：2117-2125,2016。
This study provided a new method to in vitro evaluate the biocompatibility of nanoscaled scaffolds for tissue engineering with neutrophils other than ordinary cell culture. The neutrophils were separated from human peripheral blood of healthy subjects. In vitro degradation product of nanohydroxyapatite/collagen (nHAC), nanohydroxyapatite/collagen/poly (L-lactic acid) (nHACP), and nHACP reinforced by chitin fibers (nHACP/CF) in the D-Hank's Balanced Salt Solution (D-HBSS) was used as the testing solution, which was thereafter mixed with the neutrophils. It was shown that the cell survival rate in the testing solutions had no significant difference from that in the D-HBSS (control). However, from both gene and protein expression levels, the lactate dehydrogenase and tumor necrosis factor-alpha of the neutrophils in the nHACP/CF testing solution were found lowest during the whole testing period; the main reasons of which might be that the calcium release rate of the scaffold was slowest and that the pH value of its degradation solution was nearest to that of human body. Moreover, in vivo experiments showed that most inflammation reactions happened for nHAC and poly (L-lactic acid) groups, while the least inflammation reactions happened for nHACP/CF group in the subcutaneous dorsum of mice at 2weeks after the surgery, which confirmed the in vitro findings. These results indicated that the pH value and the certain metal iron concentration of the nanoscaled scaffold degradation solution should be two important factors that significantly affect its biocompatibility. This study provides a simple and effective biocompatibility test method for biodegradable nanoscaled tissue engineering scaffolds. (c) 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2117-2125, 2016.