Effects of combinations of BMP-2 with FGF-2 and/or VEGF on HUVECs angiogenesis in vitro and CAM angiogenesis in vivo

Abstract

Angiogenesis, a complex biologic process, is regulated by a large number of angiogenic factors, including vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2). Whether Bone morphogenetic proteins-2 (BMP-2), the osteoinductive factor, could significantly reinforce the effect of VEGF and FGF-2 on angiogenesis has not been studied in detail. To study the positive effects of multiple growth factors on angiogenesis, HUVECs were treated with BMP-2, VEGF, or FGF-2 singly and in binary and ternary combinations. This study further investigates the optimal timing of the ternary combination of BMP-2, VEGF and FGF-2 for angiogenesis in the chorioallantoic membrane (FGF-2 CAM). Results of single applications of BMP-2, VEGF, or FGF-2 suggested that HUVECs angiogenesis could be promoted in a dose-dependent manner and that the optimal concentration of BMP, VEGF and FGF-2 was 10, 50 and 1 ng/mL, respectively. These results indicated that the angiogenic activity of VEGF and FGF-2 was amplified by combining with BMP-2. The ternary combination of BMP-2, VEGF and FGF-2 exhibited a positive and synergistic effect on HUVECs angiogenesis, with the lower concentrations of each factor (1 ng/mL of BMP-2, 25 ng/mL of VEGF and 0.1 ng/mL of FGF-2) being sufficient to show synergistic promotion. When VEGF and FGF-2 were added in the initial activation stage and BMP-2 was added in the maturation stage, both HUVECs angiogenesis in vitro and CAM angiogenesis in vivo could be enhanced more effectively. These results could provide a basis for the controlled release systems capable of delivering multiple factors sequentially to promote angiogenesis in tissue engineering.

The role of endoglin in post-ischemic revascularization

“…BMP2, 4, 6 and 7 have been reported to increase VEGF levels, to enhance capillary tube formation, to promote the proliferation and migration of ECs during the activation phase of angiogenesis, and to regulate the physiology of mural cells during the maturation phase [141,142]. In contrast, antiangiogenic responses have also been reported in other studies, again depending on the BMP dose and cellular context [143,144].…”

Four-week histologic evaluation of grafted calvarial defects with adjunctive hyperbaric oxygen therapy in rats

“…Therefore, in the present study, both BMP-2 and VEGF were applied to the bone defect. One previous study found that BMP-2 could amplify the angiogenic activity of VEGF [20]. However, there is an ideal and specific ratio of BMP-2:VEGF for new bone formation [21].…”

Dual Role of Mesenchymal Stem Cells Allows for Microvascularized Bone Tissue‐Like Environments in PEG Hydrogels

“…BMP-2 has previously been shown to promote angiogenesis ex vivo in mouse bones explants. [ 53 ] Moreover, in vitro BMP-2 directly activated ECs and promoted migration in transwell and scratch migration assays [ 54 ] as well as the formation of microvascular structures on matrigel. [ 55,56 ] However, in our hydrogels the stimulation with BMP-2 or FGF-2 is not suffi cient to support the formation of vascular structures by HUVECs in monocultures ( Figure S1a, Supporting Information; data not shown for BMP-2).…”