Marin-Tapia Hugo Arian, Romero-Salazar Lorena, Arteaga-Arcos Juan Carlos, Rosales-Ibáñez Raúl, Mayorga-Rojas Miguel
Rheology, Nanofluids and Microfluidics Laboratory, Autonomous University of the State of Mexico, Faculty of Sciences, Campus Universitario ''El Cerrillo, Piedras Blancas", Carretera Toluca-Ixtlahuaca km 15.5, C.P. 50200 Estado de México, Mexico; Autonomous University of the State of Mexico, Faculty of Sciences, Campus Universitario ''El Cerrillo, Piedras Blancas", Carretera Toluca-Ixtlahuaca km 15.5, C.P. 50200 Estado de México, Mexico.
Nanothermodynamics and Complex Systems Laboratory, Autonomous University of the State of Mexico, Faculty of Sciences, Campus Universitario ''El Cerrillo, Piedras Blancas", Carretera Toluca-Ixtlahuaca km 15.5, C.P. 50200 Estado de México, Mexico.
J Mech Behav Biomed Mater. 2021 Jul;119:104510. doi: 10.1016/j.jmbbm.2021.104510. Epub 2021 Apr 16.
The present study elucidates the impact of detergent-based chemical decellularization on the micro-mechanical properties of porcine and rabbit corneas for the purpose of extracellular matrix (ECM) derived scaffolds. Aiming to optimize the decellularization process, different concentrations of Sodium Dodecyl Sulfate (SDS), Triton X-100 and CHAPS detergents were assessed on their ability to decellularize corneas from both bio-models at incubation periods of 12 and 24h. We evaluated the effect of decellularization on corneal ECM Young's Modulus and various area's roughness parameters (topography features) at a microscale by using Atomic Force Microscopy (AFM). Only SDS presented adequate decellularization properties at the selected concentrations (0.2, 0.5 and 1%) and incubation periods. All topography features displayed by native corneas were preserved after SDS treatments, while no statistically significant differences were identified for the average value of Young's Modulus between the control samples and those treated with 0.2% SDS (rabbit corneas) and 0.5% SDS (porcine corneas) after 12h. In this sense, cornea decellularization procedures can be improved by simultaneously reducing SDS concentration and incubation period. AFM is a useful tool to perform biomechanical analysis of the effect of decellularization on scaffold micro-mechanics. Evaluation of the scaffold mechanical behavior at a microscale could help in understanding cell-scaffold interactions in terms of mechanotransduction, complementing macroscale techniques (e.g. tensile tests) relevant for tissue engineering quality control and decision-making.
本研究阐明了基于去污剂的化学去细胞处理对猪和兔角膜微机械性能的影响，目的是获得细胞外基质（ECM）衍生支架。为了优化去细胞处理过程，评估了不同浓度的十二烷基硫酸钠（SDS）、曲拉通X-100和3-[(3-胆酰胺丙基)二甲基铵]-1-丙磺酸（CHAPS）去污剂在12小时和24小时孵育期对两种生物模型角膜去细胞的能力。我们使用原子力显微镜（AFM）在微观尺度上评估了去细胞处理对角膜ECM杨氏模量和不同区域粗糙度参数（形貌特征）的影响。在所选择的浓度（0.2%、0.5%和1%）和孵育期下，只有SDS表现出足够的去细胞性能。SDS处理后保留了天然角膜显示的所有形貌特征，并且在12小时后，对照样品与用0.2% SDS处理的样品（兔角膜）和用0.5% SDS处理的样品（猪角膜）之间的杨氏模量平均值没有统计学上的显著差异。从这个意义上说，可以通过同时降低SDS浓度和孵育期来改进角膜去细胞程序。AFM是一种用于对去细胞处理对支架微机械性能的影响进行生物力学分析的有用工具。在微观尺度上评估支架的力学行为有助于从机械转导方面理解细胞-支架相互作用，补充与组织工程质量控制和决策相关的宏观技术（例如拉伸试验）。
