Grooved hydroxyapatite scaffold modulates mitochondria homeostasis and thus promotes osteogenesis in bone mesenchymal stromal cells

Abstract

Hydroxyapatite scaffolds (HASs) are widely studied as suitable materials for bone replacement scaffolds due to their chemical similarities to organic materials. In our previous study, a novel HAS with a 25-30-μm groove structure (HAS-G) exhibited enhanced osteogenesis of bone mesenchymal stromal cells (BMSCs) compared with HAS, potentially by modulating the macrophage-induced immune microenvironment. However, the exact effects of different surface patterns on the physiological processes of attached cells is not known. The present study aimed to determine the effects of HAS-G on the osteogenesis and physiological processes in BMSCs. Cell counting kit-8 assays and propidium iodide staining followed by flow cytometry were performed, and the results demonstrated that both in normal medium and differentiating medium, HAS-G promoted cell proliferation by decreasing the proportion of G1/G0 cells and decreased reactive oxygen species (RO S) accumulation in BMSCs compared with HAS. Detection markers of osteogenesis revealed that compared with HAS, HAS-G increased runt-related transcription factor 2, osteocalcin and osteopontin protein levels and promoted osteogenesis, which was further confirmed by Alizarin Red S staining. Following JC- 1 staining, it was observed that HAS-G maintained the mitochondrial membrane potential, similar to that achieved by N- acetylcysteine pretreatment. In addition, compared with those of HAS, HAS-G decreased mitochondrial RO S levels, which potentially contributed to the promotion of osteogenesis. The results also demonstrated that HAS-G inhibited mitophagy induced by RO S accumulation and ATP synthesis compared with HAS. In conclusion, HAS-G decreased RO S accumulation and mitophagy and thus promoted osteogenesis of BMSCs, indicating that RO S modulation of HAS-G may serve a key role in osteogenesis.