Three-dimentional cultured adipose mesenchymal stem cells for alveolar cleft bone repair: an in vitro study / 中华医学美学美容杂志

ABSTRACT

Objective:To explore the osteogenic effect of adipose-derived mesenchymal stem cells (ADSCs) in three-dimensional culture conditions in Matrigel hydrogel to provide theoretical basis for tissue engineering bone repair of alveolar process fractures.Methods:This study was performed in the Laboratory of Guangzhou Women and Children Medical Center in June 2019. The fourth generation of ADSCs were used to adjust the cell density to 1×10 5/mL. Two-dimensional common culture media (group A) and two-dimensional osteogenic induction media (group B) were used. The ADSCs were encapsulated in hydrogel with osteogenic induction media (group C), and the proliferation of cells was detected by CCK8 method. The cell mineralization effect was detected by alizarin red staining and alkaline phosphatase activity. The expression levels of osteocalcin (OCN) and osteopontin (OPN) were detected by qRT-PCR and Western blot assay. Results:ADSCs were encapsulated in hydrogel. The results of CCK8 kit showed that ADSCs could stably proliferate under the three-dimensional culture. The alizarin red staining and alkaline phosphatase activity of ADSCs under three-dimensional culture conditions were significantly higher than those of two-dimensional condition. In the detection of Western blotting and RT-PCR, we found that the expression of osteogenic protein and mRNA (OCN and OPN) of ADSCs in three-dimensional culture was also higher than that of two-dimensional culture.Conclusions:The encapsulation of ADSCs in hydrogel does not affect the proliferative potential of stem cells, and 3D cultured stem cells can significantly enhance their osteogenesis in vitro.