Liu Yushan, Chen Pei, Zhou Tengfei, Zeng Jincheng, Liu Ziyi, Wang Ruijie, Xu Yiwei, Yin Wuwei, Rong Mingdeng
Department of Periodontology and Implantology, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, Guangdong Key Laboratory of Medical Bioactive Molecular Developmental and Translational Research, Guangdong Medical University, Dongguan, China.
Front Cell Dev Biol. 2024 May 6;12:1378035. doi: 10.3389/fcell.2024.1378035. eCollection 2024.
Stem cell spheroid is a promising graft substitute for bone tissue engineering. Spheroids obtained by 3D culture of STRO1+ Gingival Mesenchymal Stem Cells (sGMSCs) (sGMSC spheroids, GS) seldom express angiogenic factors, limiting their angiogenic differentiation . This study introduced a novel stem cell spheroid with osteogenic and angiogenic potential through 3D co-culture of sGMSCs and Human Umbilical Vein Endothelial Cells (HUVECs) (sGMSC/HUVEC spheroids, GHS). GHS with varying seeding ratios of sGMSCs to HUVECs (GHR) were developed. Cell fusion within the GHS system was observed via immunofluorescence. Calcein-AM/PI staining and chemiluminescence assay indicated cellular viability within the GHS. Furthermore, osteogenic and angiogenic markers, including ALP, OCN, RUNX2, CD31, and VEGFA, were quantified and compared with the control group comprising solely of sGMSCs (GS). Incorporating HUVECs into GHS extended cell viability and stability, initiated the expression of angiogenic factors CD31 and VEGFA, and upregulated the expression of osteogenic factors ALP, OCN, and RUNX2, especially when GHS with a GHR of 1:1. Taken together, GHS, derived from the 3D co-culture of sGMSCs and HUVECs, enhanced osteogenic and angiogenic capacities , extending the application of cell therapy in bone tissue engineering.
干细胞球状体是骨组织工程中一种很有前景的移植替代物。通过STRO1+牙龈间充质干细胞（sGMSCs）的三维培养获得的球状体（sGMSC球状体，GS）很少表达血管生成因子，限制了它们的血管生成分化。本研究通过sGMSCs与人类脐静脉内皮细胞（HUVECs）的三维共培养（sGMSC/HUVEC球状体，GHS）引入了一种具有成骨和血管生成潜力的新型干细胞球状体。开发了具有不同sGMSCs与HUVECs接种比例（GHR）的GHS。通过免疫荧光观察GHS系统内的细胞融合。钙黄绿素-AM/PI染色和化学发光测定表明GHS内的细胞活力。此外，对包括碱性磷酸酶（ALP）、骨钙素（OCN）、RUNX2、CD31和血管内皮生长因子A（VEGFA）在内的成骨和血管生成标志物进行了定量，并与仅由sGMSCs组成的对照组（GS）进行了比较。将HUVECs纳入GHS可延长细胞活力和稳定性，启动血管生成因子CD31和VEGFA的表达，并上调成骨因子ALP、OCN和RUNX2的表达，尤其是当GHR为1:1的GHS时。综上所述，源自sGMSCs和HUVECs三维共培养的GHS增强了成骨和血管生成能力，扩展了细胞疗法在骨组织工程中的应用。
