Aqueous extract of Ormocarpum cochinchinense promotes fracture healing through coordinated angiogenic, osteogenic, and cytoprotective mechanisms.

Ormocarpum cochinchinense has long been used in traditional medicine for the management of bone fractures. Despite its widespread ethnomedicinal use, scientific evidence supporting its efficacy and underlying mechanisms in fracture healing remains limited. This study aimed to investigate the safety and fracture-healing potential of the aqueous extract of O. cochinchinense (OCE) with emphasis on angiogenesis and osteogenesis, and to elucidate its role in modulating endothelial cell inflammation. The biological activity of OCE was assessed using a combination of in vitro and in vivo approaches. The phytochemical characterization of the extract was performed using GC-MS analysis. The safety profile of OCE was assessed through in vivo toxicity evaluation and the fracture-healing efficacy was investigated through radiographic and histological analyses. Molecular mechanisms were examined by assessing the expression of angiogenic and osteogenic marker genes. Additionally, endothelial protective effects were evaluated under lipopolysaccharide (LPS)-induced inflammatory conditions by measuring oxidative stress markers and pro-inflammatory mediators. GC-MS analysis identified a total of 25 phytocompounds in OCE. OCE exhibited no observable toxicity in rats. It accelerated fracture healing by promoting advanced callus formation, enhancing collagen-rich matrix deposition, and improving mineralization. OCE significantly upregulated angiogenic (VEGFA, VEGFR2) and osteogenic (BMP2, SP7, RUNX2, ALP, Col1A1, OPN, OCN) gene expression, indicating coordinated activation of bone regeneration pathways. Furthermore, OCE protected endothelial cells from LPS-induced cytotoxicity by reducing reactive oxygen species and nitric oxide levels and downregulating inflammatory mediators including NOX1, NOX2, NOX4, iNOS, IL-1β, IL-6, and TNF-α. OCE promoted fracture healing through a multifaceted mechanism involving enhanced angiogenesis and osteogenesis. It also confers endothelial protection by mitigating inflammation and oxidative stress that might aid in improved endothelial function and thereby fracture healing. These findings validate its traditional use and highlight its potential as a safe and effective natural therapeutic agent for bone regeneration.