Kim Dongyun, Lee Hyeongjin, Lee Geum-Hwa, Hoang The-Hiep, Kim Hyung-Ryong, Kim Geun Hyung
Department of Biomechatronic Engineering, College of Biotechnology and Bioengineering Sungkyunkwan University (SKKU) Suwon Republic of Korea.
Non-Clinical Evaluation Center, Biomedical Research Institute Jeonbuk National University Hospital Jeonju Republic of Korea.
Bioeng Transl Med. 2022 Apr 5;7(3):e10317. doi: 10.1002/btm2.10317. eCollection 2022 Sep.
The goal of this study was to fabricate bioactive cell-laden biocomposites supplemented with bone-derived decellularized extracellular matrix (dECM) with calcium phosphate ceramic, and to assess the effect of the biocomponents on the osteogenic and odontogenic differentiation of human dental pulp stem cells (hDPSCs). By evaluating the rheological properties and selecting printing parameters, mechanically stable cell-laden 3D biocomposites with high initial cell-viability (>90%) and reasonable printability (≈0.9) were manufactured. The cytotoxicity of the biocomposites was evaluated via MTT assay and nuclei/F-actin fluorescent analyses, while the osteo/odontogenic differentiation of the hDPSCs was assessed using histological and immunofluorescent analyses and various gene expressions. Alkaline phosphate activity and alizarin red staining results indicate that the dECM-based biocomposites exhibit significantly higher osteogenic activities, including calcification, compared to the collagen-based biocomposites. Furthermore, immunofluorescence images and gene expressions demonstrated upregulation of dentin matrix acidic phosphoprotein-1 and dentin sialophosphoprotein in the dECM-based biocomposites, indicating acceleration of the odontogenic differentiation of hDPSCs in the printed biocomposites. The hDPSC-laden biocomposite was implanted into the subcutaneous region of mice, and the biocomposite afforded clear induction of osteo/odontogenic ectopic hard tissue formation 8 weeks post-transplantation. From these results, we suggest that the hDPSC-laden biocomposite is a promising biomaterial for dental tissue engineering.
本研究的目的是制备一种生物活性载细胞生物复合材料，该材料添加了骨源性脱细胞细胞外基质（dECM）和磷酸钙陶瓷，并评估这些生物成分对人牙髓干细胞（hDPSC）成骨和成牙分化的影响。通过评估流变学特性并选择打印参数，制造出了具有机械稳定性的载细胞3D生物复合材料，其初始细胞活力高（>90%）且具有合理的可打印性（≈0.9）。通过MTT试验以及细胞核/F-肌动蛋白荧光分析评估了生物复合材料的细胞毒性，同时使用组织学和免疫荧光分析以及各种基因表达来评估hDPSC的成骨/成牙分化。碱性磷酸酶活性和茜素红染色结果表明，与基于胶原蛋白的生物复合材料相比，基于dECM的生物复合材料表现出显著更高的成骨活性，包括钙化。此外，免疫荧光图像和基因表达显示，基于dECM的生物复合材料中牙本质基质酸性磷酸蛋白-1和牙本质涎磷蛋白上调，表明打印生物复合材料中hDPSC的成牙分化加速。将载有hDPSC的生物复合材料植入小鼠皮下区域，移植8周后，该生物复合材料明显诱导了成骨/成牙异位硬组织形成。基于这些结果，我们认为载有hDPSC的生物复合材料是一种有前途的牙科组织工程生物材料。
