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Vitamin D is important to normal bone and cartilage metabolism. Insufficient concentrations of vitamin D adversely affect calcium metabolism, osteoblastic activity, matrix ossification, bone density, and articular cartilage turnover. Osteoarthritis (OA) is a degenerative joint disease involving the cartilage and many of its surrounding tissues. In addition to damage and loss of articular cartilage, there is remodelling of subarticular bone, osteophyte formation, ligamentous laxity, weakening of periarticular muscles, and, in some cases, synovial inflammation. Pain is the major clinical symptom in knee OA joint. Objective: To estimate a relation between Vit.D deficiency and the severity of knee osteoarthritis. Patients and Methods: This study was a cross sectional study conducted on ninety (90) patients diagnosed (aged 40-55 years old) with knee osteoarthritis according to according to American college of rheumatology (ARC) criteria 2016 for diagnosis of OA. Patients were recruited from the Rheumatology Outpatient Clinic and Internal Medicine and Rheumatology Department at Ain Shams University Hospitals. Results: Our studied cases show that VAS pain score ranges from 3–7 with a mean 4.70 6 0.93 and median 4. As regard pain domini of WOMAC, it had significant relation with Vit D significant (p value ¼ 0.007) while VAS Pain Score not significant related to Vit D (p value ¼ 0.121). In current study, the correlation between stiffness and Vit D is statistically significant (p value ¼0.001), the correlation between physical function and Vit D is statistically significant (p value ¼ 0.001), the correlation between WOMAC score and Vit D was statistically significant (p value ¼ 0.001) As regard relation between Vit. D and X-Ray Kellgran-Lawrence system, it was significant in grade 1and grade 3. But it was non-significant n grade 2 &4. Conclusion: This study has demonstrated a significant association between knee OA and vitamin D deficiency and corresponding with the development of knee cartilage damage. As Low levels of vitamin D may not play a critical role in OA pathogenesis and progression [ABSTRACT FROM AUTHOR]