[Study on promoting bone formation in osteoporotic zebrafish by lithium-doped hydroxyapatite nanowires].

Abstract

Purpose: To observe the regulatory effect of lithium-doped hydroxyapatite nanowires on bone metabolism in osteoporotic zebrafish induced by dexamethasone.

Methods: Pure hydroxyapatite nanowires(nHA) and hydroxyapatite nanowires doped with 10% lithium ions (Li-nHA) were prepared by using hydrothermal method, and then material characterization was performed. The juvenile zebrafish cultured for 3 days(3dpf) were selected and co-cultured with nHA and Li-nHA extracts up to 7dpf. A negative(0.1% DMSO) control group was set up and transgenic zebrafish Tg(ola.sp7:nlsGFP) was used to select the best concentration for promoting bone formation. The osteoporotic zebrafish were induced by dexamethasone and incubated with nHA and Li-nHA extracts. The wild-type zebrafish was stained with alizarin red and the osteogenic differentiation was observed in transgenic zebrafish. Real-time quantitative PCR was adopted to detect osteogenic maker genes, such as zinc finger transcription factor (SP7), alkaline phosphatase (ALP), osteoprotegerin (OPG), Runt related transcription factor 2(Runx2) and osteocalcin (OCN). Statistical analysis was performed with GraphPad Prism 9.3 software.

Results: nHA and Li-nHA promoted bone formation and up-regulated expression levels of ALP, OCN, Runx2, SP7 and OPG of osteoporotic zebrafish. Compared with nHA, Li-nHA significantly increased the mineralization specific staining area and cumulative optical density of zebrafish bone, and the expression of osteogenic maker genes was also significantly increased.

Conclusions: Doping lithium ions in nano hydroxyapatite can enhance its osteoinductive properties, and Li-nHA can effectively improve bone formation of osteoporotic zebrafish.