Concentrated Growth Factors Promote hBMSCs Osteogenic Differentiation in a Co-Culture System With HUVECs

Abstract

Osteogenesis is a complex physiologic process that occurs during bone regeneration. This process requires several growth factors that act on bone marrow-derived mesenchymal stem cells (BMSCs). Concentrated growth factor (CGF) is a new-generation platelet-rich derivative that is an appealing autologous material for application in tissue repair and bone regenerative medicine because it contains a variety of fibrin and growth factors. In this study, the effects of CGF on the proliferation and osteogenic differentiation of hBMSCs and human umbilical vein endothelial cells (HUVECs) were explored with in vitro cell co-culture experiments. HBMSCs and HUVECs were directly co-cultured at the ratio of 1:2 under different concentrations (0, 2, 5, 10, 20%) of CGF for 7 days. Alkaline phosphatase (ALP) staining and quantitative reverse transcription polymerase chain reaction were used to detect the effects of CGF on the expression of osteogenic genes (ALP, osteocalcin [OCN], type I collagen [COL-1], Runt-related transcription factor 2 [RUNX2]) and connexin 43 (CX43). RNA sequencing was used to explore potential regulated genes and signaling pathways that affect the osteogenesis of co-cultured hBMSCs exposed to CGF. The results showed higher expressions of ALP, COL-1, RUNX2, OCN, and CX43 in the direct co-culture group containing 10% CGF compared to the direct co-culture group without CGF and the indirect co-culture group. In summary, 10% CGF can significantly promote osteogenesis in hBMSCs directly co-cultured with HUVECs. Intercellular communication between the direct co-culture of hBMSCs and HUVECs through CX43 may be one of the main regulatory mechanisms.

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“…Currently, ALP , Runx2 , OCN (osteocalcin), and COL-I have been widely recognized as specific markers for osteogenic differentiation. 28,29 The introduction of specific biomolecules can promote bone healing. Boden et al 30 demonstrated in clinical studies that BMP2 has a promoting effect on bone formation in spinal fusion.…”

BMSC–HNC Interaction: Exploring Effects on Bone Integrity and Head and Neck Cancer Progression

“…The study found that 10% concentrated growth factor (CGF) had a significant osteogenic effect on human bone marrow mesenchymal stem cells (hBMSCs) directly co-cultured with human umbilical vein endothelial cells (HUVECs). The researchers suggest that autophagy activated by the mTOR/ULK1 pathway and CX43-mediated intercellular communication may play regulatory roles in promoting hBMSCs osteogenic differentiation [29].…”

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