Background: Metabolic syndrome (MetS) is considered as common risk factors for knee osteoarthritis (OA). Aim of Study: Comparing the effect of vit. D supplementation and gut microbiota suppression on the development of metabolic knee OA in rats is the aim of the study. Material and Methods:Forty adult male rats were subdivided into control and high carbohydrate high fat (HCHF) fed rats that furtherly subdivided into: HCHF (positive control), HCHF-Vitamin D treated & HCHF-antibiotics treated. Body mass index (BMI), serum glucose, insulin, total cholesterol, triglycerides, HDL-C, LDL-C, inflammatory cytokines were estimated. Homeostatic model assessment for insulin resistance (HOMA-IR) was calculated. Synovial cytokine, tissue degenerative enzymes gene expression and knee histopathology were assessed. Results: HCHF induced significant increase in BMI, glucose, insulin, HOMA-IR, serum and synovial cytokines and degenerative enzymes with dyslipidemia and knee joint damage compared with controls. Vit. D improved all parameters in comparison to HCHF fed group. Antibiotic administration improved glucose, insulin, HOMA-IR, dyslipidemia and serum cytokines but induced insignificant changes in BMI, synovial cytokines, degenerative enzymes and knee joint damage compared with HCHF fed group. Conclusion: HCHF diet led to development of knee joint damage associated with changes in the metabolic profile in rats. Vit D supplementation dramatically improved knee joint damage, obesity insulin resistance, dyslipidemia and inflammation systemically and locally while controlling gut dysbiosis by antibiotics did not improve obesity, local inflammation and knee damage but improved the other parameters less than Vit D