Osteoblastogenesis, Osteolysis, and Adipogenesis in the Male Diabetic Sprague Dawley Rat Mandible Treated with Antiretroviral Therapy and Chronic Alcohol

Background: Diabetes mellitus (DBT), alcohol (ALC) consumption, and antiretroviral therapy (cART) are among the chronic conditions known to affect bone turnover, reduce osteoblastogenesis, increase osteolysis, and increase susceptibility to fracture. The combined deleterious effects of these conditions on bone cells (adipocytes and osteocytes) and the expression pro- and anti-osteogenic cytokines (TGF-β1 and BMP-3) have not been elucidated in the scientific literature. Aim: This study aimed to assess the influence of DBT, ALC, and cART on adipocytes, osteocytes and the expression of TGF-β1 and BMP-3 in the mandibular bone of male diabetic Sprague Dawley rats (SD). Methods: This study involved seventy-two (72) male adult SD rats that were respectively grouped as untreated control (n=8), ALC (n=8), DBT (n=7), cART (n=8), rats that received both ALC and cART (ALC+cART) (n=7), DBT rats that received ALC (DBT+ALC) (n=6), DBT rats that received cART (DBT+cART) (n=6), DBT rats that received both ALC and cART (DBT+ALC+cART)(n=6), the Gel (n=8), and the buffered citrate (n=8). The rats that received ALC were given 10 % v/v alcohol daily in drinking water for 12 weeks. The cART was given to the treated rats with an extrapolated adult dose of 0.023mg/kg B.W. of cART daily. Diabetes was induced through a 20% fructose diet and one-off intraperitoneal injection of 40mg/kg B.W. of STZ, and was confirmed through low serum insulin level, OGTT level, and FBG levels exceeding 250mg/dL. After the 12 ^th week, mandibular bones were harvested and fixed in 10% buffered formalin, before decalcification in EDTA pH 7.4. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 µm thickness in a series for H&E, and IHC was performed with TRAP, BMP-3, and TGF-β1 antibodies. Results: Alcohol intake (ALC) or cART or a combination (ALC) and (cART) (ALC+cART) or diabetes (DBT) or concurrent use of ALC and cART among diabetics (DBT+ALC+cART) increased bone marrow adiposity. The inhibitory effect of BMP-3 was coupled with a reduction in osteocyte quantities (TRAP immunopositive cells) without a significant change in the expression of TGF-β1. Conclusion: The results showed that when DBT was confounded by cART and ALC (DBT+ALC+cART), osteogenic cells were negatively affected, demonstrated by increased adipogenesis, reduced osteoblastogenesis, and increased osteolysis.