Matrix alteration and not residual sodium dodecyl sulfate cytotoxicity affects the cellular repopulation of a decellularized matrix
Matrix alteration and not residual sodium dodecyl sulfate cytotoxicity affects the cellular repopulation of a decellularized matrix
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DOI：
10.1089/ten.2006.12.2975
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发表时间：
2006-10-01
影响因子：
--
通讯作者：
Woods, Terence
中科院分区：
文献类型：
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作者：
Gratzer, Paul F.;Harrison, Robert D.;Woods, Terence
有人提出，残留的细胞毒性十二烷基硫酸钠（SDS）是导致在SDS脱细胞组织中观察到的细胞向内生长水平较低的原因。为了确定是否如此，我们使用了两种洗涤方法来去除残留的SDS，并进行了广泛的生化、力学和结构分析，以确定基于SDS的脱细胞处理对猪前交叉韧带（ACL）组织的影响及其细胞再植入的倾向。
使用两种不同的洗涤技术（pH = 9的缓冲液、75%乙醇）降低了脱细胞组织中残留的SDS水平。在pH = 9或75%乙醇中洗涤后，发现脱细胞组织中残留的SDS浓度分别比所报道的SDS细胞毒性水平低约8倍和23倍。研究发现，SDS处理显著降低了糖胺聚糖水平，增加了胶原卷曲幅度和周期性，并增加了胶原被明胶酶胰蛋白酶降解的敏感性。在培养28天后，无论洗涤技术如何以及组织中残留的SDS水平如何，脱细胞组织中自体ACL成纤维细胞的再植入水平和活力都相同。这有力地表明，SDS处理导致的组织基质生化或结构的改变，而非残留的SDS细胞毒性，是SDS脱细胞组织中观察到的细胞再植入水平较低的原因。
It has been suggested that residual cytotoxic sodium dodecyl sulfate (SDS) is responsible for the low levels of cell in-growth observed in SDS decellularized tissues. To determine whether this is the case, we used 2 washing methods to remove residual SDS and extensive biochemical, mechanical, and structural analyses to determine the effects of SDS-based decellularization on porcine anterior cruciate ligament (ACL) tissue and its propensity for cellular repopulation.The level of residual SDS in decellularized tissue was reduced using 2 different washing techniques (pH = 9 buffer, 75% ethanol). After washing in pH = 9 or 75% ethanol, residual SDS concentrations in decellularized tissues were found to be approximately 8 and 23 times less than reported SDS cytotoxic levels, respectively. It was found that SDS treatment significantly reduced glycosaminoglycan levels, increased collagen crimp amplitude and periodicity, and increased susceptibility of collagen to degradation by the gelatinase enzyme trypsin. The level of repopulation and viability of autologous ACL fibroblasts in the decellularized tissue after 28 days of culture were found to be the same regardless of the washing technique and resulting level of residual SDS in the tissue. This strongly indicates that alterations in tissue matrix biochemistry or structure from SDS treatment and not residual SDS cytotoxicity are responsible for the low cell re-population observed in SDS decellularized tissues.