Abstract：
The decellularized matrix has a great potential for tissue remodeling and regeneration; however, decellularization could induce host immune rejection due to incomplete cell removal or detergent residues, thereby posing significant challenges for its clinical application. Therefore, the selection of an appropriate detergent concentration, further optimization of tissue decellularization technique, increased of biosafety in decellularized tissues, and reduction of tissue damage during the decellularization procedures are pivotal issues that need to be investigated. In this study, we tested several conditions and determined that 0.1% Sodium dodecyl sulfate and three decellularization cycles were the optimal conditions for decellularization of pulp tissue. Decellularization efficiency was calculated and the preparation protocol for dental pulp decellularization matrix (DPDM) was further optimized. To characterize the optimized DPDM, the microstructure, odontogenesis-related protein and fiber content were evaluated. Our results showed that the properties of optimized DPDM were superior to those of the non-optimized matrix. We also performed the 4D-Label-free quantitative proteomic analysis of DPDM and demonstrated the preservation of proteins from the natural pulp. This study provides a optimized protocol for the potential application of DPDM in pulp regeneration.
摘要：
脱细胞基质具有组织重塑和再生的巨大潜力；然而，去细胞化可以诱导宿主免疫排斥由于不完全的细胞去除或洗涤剂残留，从而对其临床应用提出了重大挑战。因此,选择合适的洗涤剂浓度，进一步优化组织脱细胞技术，脱细胞组织的生物安全性增加，在去细胞化过程中减少组织损伤是需要研究的关键问题。在这项研究中，我们测试了几种条件，并确定0.1％十二烷基硫酸钠（SDS）和三个脱细胞周期是牙髓组织脱细胞的最佳条件。计算了脱细胞效率，并进一步优化了牙髓脱细胞基质（DPDM）的制备方案。要表征优化的DPDM，微观结构，评估牙本质形成相关蛋白和纤维含量。我们的结果表明，优化的DPDM的性能优于未优化的基体。我们还进行了DPDM的4D无标签定量蛋白质组学分析，并证明了天然纸浆中蛋白质的保存。该研究为DPDM在纸浆再生中的潜在应用提供了坚实的理论和实验基础。
