Koh Benson, Ab Rahman Farynna Hana, Matlan Najwa Amira, Rajan Manissha, Musta'ain Aimi Yasmin, Mohd Jeffry Lee Mohamad Ridhwan, Ramli Roszalina, Mohd Yunus Siti Salmiah, Binti Hj Idrus Ruszymah, Yazid Muhammad Dain
Centre for Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Cheras, 56000, Kuala Lumpur, Malaysia.
Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia.
Biol Res. 2022 Mar 4;55(1):11. doi: 10.1186/s40659-022-00380-8.
Functional bioengineered tooth regeneration using autologous or allogeneic alternative differentiated cells sources are thought to have a great potential in replacing conventional dentures. This study investigated the potential of dental pulp stem cells (DPSCs) conditioned medium for odontoblastic differentiation of Wharton's jelly mesenchymal stem cells (WJMSCs). The DPSCs derived from healthy adult permanent first molars were cultured at high confluence prior to conditioned medium collection. The WJMSCs were cultured in six different treatments, with varying ratios of culture media to DPSCs-conditioned medium. MTT assay was used to measure the rate of proliferation of WJMSCs, while immunocytochemistry staining was utilised to detect the expression of dental matrix protein 1 (DMP-1). The deposited calcium was detected and analysed via Alizarin-Red Staining (ARS).
It was found that the proliferation of WJMSCs cultured under the mixture of complete medium and DPSCs conditioned medium showed significantly lower than the control; presumably the cells started to exit proliferative state prior differentiation. In 14 days of induction, the cells in all treatments showed osteoblastic-like morphology, calcium compound deposits were observed at day 7, 10 and 14 of differentiation suggested that DPSCs conditioned medium could lead to osteoblastic/odontoblastic differentiation. However, the DMP-1 protein can be seen only expressed minimally at day 14 of conditioned medium induction.
In conclusion, DPSCs conditioned medium appeared as a potential odontoblastic induction approach for WJMSCs. To further investigate the stimulatory effects by DPSCs conditioned medium, specific signalling pathway need to be elucidated to enhance the differentiation efficiency.
使用自体或异体替代分化细胞来源进行功能性生物工程牙齿再生被认为在替代传统假牙方面具有巨大潜力。本研究调查了牙髓干细胞（DPSCs）条件培养基对沃顿胶间充质干细胞（WJMSCs）向成牙本质细胞分化的潜力。从健康成人恒牙第一磨牙中分离得到的DPSCs在收集条件培养基前进行高汇合培养。将WJMSCs培养于六种不同处理中，培养基与DPSCs条件培养基的比例各不相同。采用MTT法检测WJMSCs的增殖率，同时利用免疫细胞化学染色检测牙基质蛋白1（DMP-1）的表达。通过茜素红染色（ARS）检测并分析沉积的钙。
发现完全培养基与DPSCs条件培养基混合培养的WJMSCs增殖率显著低于对照组；推测细胞在分化前开始退出增殖状态。诱导14天时，所有处理组的细胞均呈现成骨样形态，在分化第7、10和14天观察到钙化合物沉积，表明DPSCs条件培养基可导致成骨/成牙本质细胞分化。然而，仅在条件培养基诱导第14天时可观察到DMP-1蛋白有少量表达。
总之，DPSCs条件培养基似乎是一种诱导WJMSCs向成牙本质细胞分化的潜在方法。为进一步研究DPSCs条件培养基的刺激作用，需要阐明特定的信号通路以提高分化效率。
