Effect of low-temperature ethylene oxide and electron beam sterilization on the in vitro and in vivo function of reconstituted extracellular matrix-derived scaffolds.
Effect of low-temperature ethylene oxide and electron beam sterilization on the in vitro and in vivo function of reconstituted extracellular matrix-derived scaffolds.
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DOI：
10.1177/0885328215590967
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发表时间：
2015-10
影响因子：
2.9
通讯作者：
Murray MM
中科院分区：
文献类型：
--
作者：
Proffen BL;Perrone GS;Fleming BC;Sieker JT;Kramer J;Hawes ML;Murray MM
重组细胞外基质（ECM）衍生的支架通常用于临床前组织工程研究，作为细胞和生长因子的递送载体。转化为临床应用需要确定一种有效去除细菌但不损害支架功能的灭菌方法。为了确定灭菌效果以及对ECM支架完整性和功能的影响，对低温环氧乙烷和15kGy电子束辐照技术进行了评估。根据美国药典第71章对支架无菌性进行评估。采用酶抗性试验和凝胶电泳法测定支架基质的体外降解。测试了支架的弹性模量、屈服应力和倒塌模量。最后，对14头约克郡猪进行了ACL横断和生物增强ACL修复。在4周、6周和8周时比较韧带、滑膜和淋巴结的组织学反应。环氧乙烷和电子束辐照产生无菌支架。环氧乙烷改变支架基质后，电子束辐照后支架抗酶消化能力和蛋白质完整性略有下降。电子束处理后支架弹性模量和屈服应力增大，环氧乙烷处理后支架坍塌模量增大。两种末端灭菌技术均未检测到ACL尺寸、体内支架吸收率或滑膜、韧带和淋巴结的组织学反应的显著变化。总之，本研究确定了两种最终灭菌ECM支架的方法。体外支架的性能略有改变，但对体内周围组织的生物学反应没有显著影响。这是将新的组织工程策略转化为临床试验的关键一步。
Reconstituted extracellular matrix (ECM) -derived scaffolds are commonly utilized in preclinical tissue engineering studies as delivery vehicles for cells and growth factors. Translation into clinical use requires identifying a sterilization method that effectively removes bacteria but doesn’t harm scaffold function. To determine effectiveness of sterilization and impact on ECM scaffold integrity and function low temperature ethylene oxide and 15kGy electron beam irradiation techniques were evaluated. Scaffold sterility was assessed in accordance to United States Pharmacopeia Chapter 71. Scaffold matrix degradation was determined in vitro using enzymatic resistance tests and gel electrophoresis. Scaffold mechanics including elastic modulus, yield stress and collapse modulus were tested. Lastly, 14 Yorkshire pigs underwent ACL transection and bio-enhanced ACL repair using sterilized scaffolds. Histologic response of ligament, synovium and lymph nodes was compared at 4, 6, and 8 weeks. Ethylene oxide as well as electron beam irradiation yielded sterile scaffolds. Scaffold resistance to enzymatic digestion and protein integrity slightly decreased after electron beam irradiation while ethylene oxide altered scaffold matrix. Scaffold elastic modulus and yield stress were increased after electron beam treatment, while collapse modulus was increased after ethylene oxide treatment. No significant changes in ACL dimensions, in vivo scaffold resorption rate, or histologic response of synovium, ligament and lymph nodes with either terminal sterilization technique were detectable. In conclusion, this study identifies two methods to terminally sterilize an ECM scaffold. In vitro scaffold properties were slightly changed without significantly influencing the biologic responses of the surrounding tissues in vivo. This is a critical step toward translating new tissue engineering strategies to clinical trials.