Effect of α2-macroglobulin in the early stage of jaw osteoradionecrosis

Advanced osteoradionecrosis (ORN) is one of the most serious complications in patients with head and neck cancer, resulting in poor prognosis. Numerous studies have therefore focused on the pathogenesis and interventions of ORN early stage. The present study aimed to investigate whether alpha 2-macroglobulin (alpha 2M) could prevent early-stage jaw osteoradionecrosis caused by radiotherapy (RT). Following local injection of alpha 2M, a single dose of 30 Gy was delivered to rats for pathological exploration. For 28 days, the irradiated mandible and soft tissues were examined for potential changes. Furthermore, primary human bone marrow mesenchymal stem cells pretreated with alpha 2M followed by 8 Gy irradiation (IR) were also used. Tartrate-resistant acid phosphatase assay, terminal uridine deoxynucleotidyl nick end labeling assay and immunohistochemical staining were performed on irradiated mandibular bone, tongue or buccal mucosa tissues from rats. Cell proliferation was assessed by evaluating the cell morphology by microscopy and by using the cell counting kit-8. Fluorescence staining, flow cytometry and western blotting were conducted to detect the reactive oxygen species level, cell apoptosis and protein expression of superoxide dismutase 2 (SOD2), heme oxygenase-1 (HO-1) and phosphorylated Akt following irradiation. The results demonstrated that alpha 2M attenuated physical inflammation, osteoclasts number and fat vacuole accumulation in mandibular bone marrow and bone marrow cell apoptosis following IRin vivo. Furthermore, alpha 2M pretreatment suppressed the expression of 8-hydroxy-2 '-deoxyguanosine in mandibular bone and tongue paraffin embedded sections, which is a marker of oxidative damage, and increased SOD2 expression in mucosa and tongue paraffin embedded sections. The present study demonstrated the efficient regulation of antioxidative enzymes, including SOD2 and heme oxygenase-1, and reduction in oxidative damage by alpha 2M. In addition,in vitroresults confirmed that alpha 2M may protect cells from apoptosis and suppress reactive oxygen species accumulation. Overall, the present study demonstrated that alpha 2M treatment may exert some radioprotective effects in early-stage ORN via antioxidant mechanisms, and may therefore be considered as a potential alternative molecule in clinical prophylactic treatments.