ACS Biomater Sci Eng. 2022 Jun 13;8(6):2574-2588. doi: 10.1021/acsbiomaterials.2c00034. Epub 2022 Jun 1.
The rise of tissue-engineered biomaterials has introduced more clinically translatable models of disease, including three-dimensional (3D) decellularized extracellular matrix (dECM) hydrogels. Specifically, decellularized nerve hydrogels have been utilized to model peripheral nerve injuries and disorders ; however, there lacks standardization in decellularization methods. Here, rat sciatic nerves of varying preparations were decellularized using previously established methods: sodium deoxycholate (SD)-based, 3-((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate (CHAPS)-based, and apoptosis-mediated. These nerves were characterized for cellular debris removal, ECM retention, and low cytotoxicity with cultured Schwann cells. The best preparations of each decellularization method were digested into dECM hydrogels, and rheological characterization, gelation kinetics, and confocal reflectance imaging of collagen fibril assembly were performed. It was determined that the SD-based method with nerve epineurial removal best maintained the overall ECM composition and mechanical properties of physiological peripheral nerves while efficiently stripping the scaffolds of tissue-specific cells and debris. This method was then utilized as a culture platform for quiescent Schwann cells and cancer-nerve crosstalk. Hydrogel-embedded Schwann cells were found to have high viability and act in a more physiologically relevant manner than those cultured in monolayers, and the hydrogel platform allowed for the activation of Schwann cells following treatment with cancer secreted factors. These findings establish a standard for peripheral nerve decellularization for usage as a dECM hydrogel testbed for peripheral nerve disease modeling and may facilitate the development of treatments for peripheral nerve disease and injury.
组织工程生物材料的兴起引入了更具临床转化潜力的疾病模型，包括三维（3D）脱细胞细胞外基质（dECM）水凝胶。具体来说，脱细胞神经水凝胶已被用于模拟周围神经损伤和疾病；然而，脱细胞方法缺乏标准化。在这里，使用先前建立的方法对不同制备的大鼠坐骨神经进行脱细胞处理：基于去氧胆酸钠（SD）、3-（（3-胆酰胺丙基）二甲氨基）-1-丙磺酸钠（CHAPS）和凋亡介导的方法。用培养的施万细胞对这些神经进行细胞碎片去除、ECM 保留和低细胞毒性的特性分析。对每种脱细胞方法的最佳制备物进行消化，得到 dECM 水凝胶，并对胶原纤维组装的流变特性、凝胶动力学和共聚焦反射成像进行分析。结果表明，具有神经外膜去除功能的 SD 基方法最能保持生理周围神经的整体 ECM 组成和机械性能，同时有效地去除支架中的组织特异性细胞和碎片。然后，该方法被用作静止施万细胞和癌-神经串扰的培养平台。发现水凝胶包埋的施万细胞具有较高的活力，并且比在单层培养中的施万细胞更能以生理相关的方式发挥作用，并且水凝胶平台允许在施万细胞接受癌症分泌因子处理后被激活。这些发现为周围神经脱细胞建立了一个标准，可作为 dECM 水凝胶的测试平台，用于周围神经疾病建模，并且可能有助于周围神经疾病和损伤的治疗方法的发展。
