Photobiomodulation therapy at 650 nm enhances osteogenic differentiation of osteoporotic bone marrow mesenchymal stem cells through modulating autophagy.

BACKGROUND

Photobiomodulatiom therapy (PBMT) has biostimulatory effects on bone marrow mesenchymal stem cells (BMSCs), which takes a pivotal role in maintaining bone mass and avoiding osteoporosis (OP). Autophagy is an important regulator for cell survival and homeostasis. Previous researchers found that BMSCs derived from osteoporotic rats (OP-BMSCs) were with the feature of reduced osteogenic differentiation and autophagy dysfunction. However, the potential regulation of PBMT in osteogenic differentiation of OP-BMSCs and its underling relationship with autophagy remain unclear.

METHODS

650 nm red light-emitting diode (LED) was selected to initiate PBMT effects. The isolation and culture of OP-BMSCs were implemented after the establishment of the OP rat model. Firstly, the optimal dose of LED was screened on OP-BMSCs by CCK-8. Meanwhile, the osteogenic and mineralization activities were studied through the detection of Alkaline phosphatase (ALP) and alizarin red S (ARS). Then, the levels of osteogenesis and autophagy were investigated via western blot and immunofluorescence staining. Finally, the autophagy inhibitor 3-MA was applied to illustrate the underlying mechanism of the osteogenic effect of PBMT on OP-BMSCs.

RESULTS

Firstly, the optimal dose of 6 J/cm LED was selected in the subsequent experiments according to CCK-8. Then, the ALP activity and the mineralization ability of OP-BMSCs were obviously increased by PBMT. Meanwhile, Runx-2, OCN and OPN were significantly upregulated in LED group. Furthermore, the expressions of autophagic proteins increased significantly in LED group by immunofluorescence staining and western blot assay. At last, the promoted effects of PBMT on osteogenic differentiation in OP-BMSCs were distinctly reversed via inhibiting autophagy.

CONCLUSION

Our research illustrated that 650 nm LED could improve osteogenic differentiation of OP-BMSCs, suggesting a potential correlation between PBMT-mediated activation of autophagy and promotion of osteogenic differentiation.