Notoginsenoside R1 facilitates stem cell-based bone tissue engineering via inducing osteogenesis, angiogenesis and cell adhesion

摘要

Cell adhesion, osteogenesis, and angiogenesis are essential phenomena for effective stem cell-based bone tissue engineering. Growth factors BMP2 and VEGF promote osteogenesis and angiogenesis, but their clinical application is still controversial due to the adverse effects and various shortcomings. Notoginsenoside R1 (NGR1) is a natural triterpene saponin compound extracted from traditional Chinese herb Panax notoginseng. Since NGR1 has high bioactivity, minimum adverse effects, and osteogenic potential, it could be used in bone regenerative application. This study aimed to investigate the effect of NGR1 on osteogenic/angiogenic differentiation and cell-adhesion of precursor cells, as well as bone regeneration during stem cell-based bone tissue engineering. We used 0-5 μg/ml of NGR1 to analyze the osteogenic and angiogenic differentiation of hASCs and HUVECs, respectively. Osteogenic/ angiogenic gene expression, ALP activity, alizarin red matrix mineralization, and matrigel-based tube formation assay were performed in vitro. Western blot analysis tested the expression of p38 and JNK. NGR1 pretreated hASCs were seeded on 3D-printed TCP scaffold, and cell-adhesion in vitro and bone regeneration in the ectopic site of nude mice were analyzed. The animal study was approved by the ethical committee of Guangzhou Medical University. The selected significance level was Pb 0.05. NGR1 promoted OCN expression, ALP activity, and matrix mineralization in dose-dependent manner, aswell asactivated the p38and JNK signalingpathway. Higher numbers of the tube-like structures were observed in NGR1 treated hASCs or HUVECs matrigel culture. Actin staining revealed the increased adhesion of NGR1-pretreated hASCs to a glass slide and TCP-scaffold. Micro-CT analysis revealed the more bone formation in ectopically grafted NGR1-pretreated hASCs-seeded 3D-printed TCP- scaffolds. In conclusion, NGR1 enhanced osteogenic/angiogenic and cell-adhesion potential of precursor cells, as well as ectopic bone formation in hASCs-loaded 3D-printed TCP-scaffolds, suggesting it as a possible therapeutic agent to enhance the efficacy of stem cell-based bone tissue engineering.