Chronic Alcohol and Combination Antiretroviral Therapy Exacerbate Mandibular Bone Deterioration via Cytokine Dysregulation in Male Diabetic Sprague DawleyRats

Objectives: Diabetes mellitus (DBT), chronic alcohol (ALC) consumption, and combination antiretroviral therapy (cART) can impair bone turnover, but their combined effects on mandibular bone cells and osteogenic cytokines remain unclear. This study evaluated these effects in male diabetic Sprague-Dawley rats. Materials and Methods: Fifty-eight 10-week-old male rats were assigned to ten groups: control, ALC, DBT, cART, ALC+cART, DBT+ALC, DBT+cART, DBT+ALC+cART, gelatin (GEL), and buffered citrate (CB). ALC was administered as 10% v/v ethanol, and cART at 0.023 mg/kg daily for 12 weeks. Diabetes was induced with a 20% fructose diet and one Streptozotocin (STZ) injection (40 mg/kg), then confirmed by fasting blood glucose (>250 mg/dL) and serum insulin. We collected mandibles, fixed and decalcified them, and assessed them using haematoxylin and eosin (H&E) staining and immunohistochemistry for tartrate-resistant acid phosphatase, bone morphogenetic protein3, and transforming growth factor-β1 (TRAP, BMP-3, and TGF-β1). Results: ALC, cART, DBT, or their combinations increased bone marrow adiposity. BMP-3 inhibition reduced osteocyte numbers (TRAP-positive cells) without significantly changing TGF-β1 expression. Conclusions: Diabetic conditions compounded by alcohol and cART exposure exacerbate bone deterioration through increased adipogenesis, reduced osteoblastogenesis, and enhanced osteolysis. These findings provide a biological basis for further investigating the oral effects of combined DBT, ALC, and cART on bone healing and implant osseointegration. Keywords: Type-2 diabetes mellitus, Alcoholism, Combination antiretroviral therapy, Mandibular bone cells, Cytokines, Osteoblastogenesis, Osteolysis