Lin Kuan-Ming, Chen Yong-Ren, Ho Ming-Hua, Li Chung-Hsing
Graduate Institute of Dental Science, National Defense Medical Center, Taipei, Taiwan.
Non-invasive Cancer Therapy Research Institute - Taiwan, Taipei, Taiwan.
J Dent Sci. 2025 Jul;20(3):1605-1614. doi: 10.1016/j.jds.2025.02.019. Epub 2025 Mar 6.
BACKGROUND/PURPOSE: Extracellular matrix (ECM) may be useful as a natural scaffold for tissue engineering. Since dental follicle stem cells (DFSCs) share a similar embryonic origin with corneal keratocytes and endothelial cells and possess the ability to proliferate and differentiate, in this study, we investigate the potential of DFSCs to differentiate into corneal cells under the supercritical carbon dioxide (SC-CO) decellularized porcine corneal ECM culture stimulation.
DFSCs were seeded onto the SC-CO decellularized porcine corneal ECM and the surrounding culture dish for 21 days. Cell morphology, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and immunofluorescence staining were used to evaluate the potential of DFSCs to differentiate into corneal epithelial, keratocyte, or endothelial cells. The culture medium was collected every 3 days for growth factor analysis.
The DFSCs cultured on the ECM exhibit notable cell-ECM adhesion and the potential to primarily differentiate into keratocytes, with a partial capacity for differentiation into corneal endothelial cells, while the cells outside the ECM are weakly affected. Growth factor array analysis revealed changes in growth factor expression in different culture conditions, DFSCs, ECM, and DFSCs/ECM co-culture at various time points, indicating a dynamic shift in the microenvironment.
The present study demonstrated that DFSCs cultured on the porcine corneal ECM can differentiate mainly into keratocytes-like and partially into corneal endothelial-like cells, exhibiting strong cell-ECM adhesion and regulation of growth factors. Provide insights into the therapeutic application potential of dental-derived stem cells and SC-CO decellularized tissue for corneal diseases.
背景/目的：细胞外基质（ECM）可作为组织工程的天然支架。由于牙囊干细胞（DFSCs）与角膜基质细胞和内皮细胞具有相似的胚胎起源，且具有增殖和分化能力，因此在本研究中，我们探讨了在超临界二氧化碳（SC-CO）脱细胞猪角膜ECM培养刺激下DFSCs分化为角膜细胞的潜力。
将DFSCs接种到SC-CO脱细胞猪角膜ECM及周围培养皿中培养21天。采用细胞形态学、逆转录定量聚合酶链反应（RT-qPCR）和免疫荧光染色评估DFSCs分化为角膜上皮细胞、基质细胞或内皮细胞的潜力。每3天收集培养基进行生长因子分析。
在ECM上培养的DFSCs表现出显著的细胞-ECM黏附能力，主要有分化为基质细胞的潜力，部分具备分化为角膜内皮细胞的能力，而ECM外的细胞受影响较弱。生长因子阵列分析显示，在不同培养条件下，DFSCs、ECM以及DFSCs/ECM共培养在各个时间点的生长因子表达均有变化，表明微环境存在动态变化。
本研究表明，在猪角膜ECM上培养的DFSCs主要可分化为类基质细胞，部分可分化为类角膜内皮细胞，表现出较强的细胞-ECM黏附能力和生长因子调控能力。为牙源性干细胞和SC-CO脱细胞组织在角膜疾病治疗中的应用潜力提供了见解。
