Division of Oral Biology, School of Dentistry, University of Leeds, Leeds, UK.
Department of Clinical Dental Science, Princess Nourah Bint Abdulrahman University, Riyadh, Kingdom of Saudi Arabia.
Sci Rep. 2020 Dec 9;10(1):21553. doi: 10.1038/s41598-020-78477-x.
Implementing the principles of tissue engineering within the clinical management of non-vital immature permanent teeth is of clinical interest. However, the ideal scaffold remains elusive. The aim of this work was to assess the feasibility of decellularising rat dental pulp tissue and evaluate the ability of such scaffold to support stem cell repopulation. Rat dental pulps were retrieved and divided into control and decellularised groups. The decellularisation protocol incorporated a low detergent concentration and hypotonic buffers. After decellularisation, the scaffolds were characterised histologically, immunohistochemistry and the residual DNA content quantified. Surface topography was also viewed under scanning electron microscopy. Biocompatibility was evaluated using cytotoxicity assays utilising L-929 cell line. Decellularised scaffolds were recellularised with human dental pulp stem cells up to 14 days in vitro. Cellular viability was assessed using LIVE/DEAD stain kit and the recellularised scaffolds were further assessed histologically and immunolabelled using makers for odontoblastic differentiation, cytoskeleton components and growth factors. Analysis of the decellularised scaffolds revealed an acellular matrix with histological preservation of structural components. Decellularised scaffolds were biocompatible and able to support stem cell survival following recellularisation. Immunolabelling of the recellularised scaffolds demonstrated positive cellular expression against the tested markers in culture. This study has demonstrated the feasibility of developing a biocompatible decellularised dental pulp scaffold, which is able to support dental pulp stem cell repopulation. Clinically, decellularised pulp tissue could possibly be a suitable scaffold for use within regenerative (reparative) endodontic techniques.
将组织工程学原理应用于非活力性未成熟恒牙的临床管理具有临床意义。然而，理想的支架仍然难以捉摸。本研究旨在评估脱细胞大鼠牙髓组织的可行性，并评估该支架支持干细胞再植的能力。从大鼠牙髓中分离出组织并分为对照组和脱细胞组。脱细胞方案采用低去污剂浓度和低渗缓冲液。脱细胞后，对支架进行组织学、免疫组织化学和残留 DNA 含量的定量分析。扫描电子显微镜观察表面形貌。利用 L-929 细胞系进行细胞毒性测定评估生物相容性。将人牙髓干细胞再植入脱细胞支架中，体外培养 14 天。使用 LIVE/DEAD 染色试剂盒评估细胞活力，并进一步通过组织学和免疫标记对再植入的支架进行评估，标记物用于牙本质分化、细胞骨架成分和生长因子。脱细胞支架的分析显示出具有结构成分的无细胞基质的保留。脱细胞支架具有生物相容性，并能够在再植入后支持干细胞的存活。再植入支架的免疫标记显示在培养中针对测试标记物的阳性细胞表达。本研究表明，开发一种生物相容性的脱细胞牙髓支架是可行的，该支架能够支持牙髓干细胞的再植。临床上，脱细胞牙髓组织可能是再生（修复）性牙髓治疗技术中合适的支架。
