Background and AimsThe primary objective of this systematic review and meta‐analysis was to assess the impact of dextrose prolotherapy on individuals diagnosed with knee osteoarthritis (KOA).MethodsTo conduct a thorough investigation, a variety of leading international databases were checked, including PubMed (Medline), Scopus, Web of Sciences, EMBASE (Elsevier), ClinicalTrials.gov, and the Cochrane Library. The search covered a period from January 2000 to the end of June 2023, which facilitated the collection of relevant studies.ResultsThe findings of the study revealed that when the studies utilizing the Western Ontario McMaster Universities Index tool (WOMAC) were combined, patients with KOA who received prolotherapy experienced an improvement in function compared with those who received other treatments (SMD: 0.20; 95% Confidence Interval [1]: −0.11, 0.51; p value SMD = 0.221; I2: 78.49%; pheterogeneity < 0.001). Additionally, there was a decrease in mean pain and stiffness among patients who received prolotherapy compared with those who received other treatments or a placebo [(SMD: −0.95; 95% CI: ‐1.14, −0.76; p value SMD < 0.001; I2: 59.35%; pheterogeneity = 0.070) and (SMD: −0.21; 95% CI: −0.32, −0.10; p value SMD < 0.001; I2: 88.11%; pheterogeneity < 0.001)]. Furthermore, based on the Visual Analog Scale (VAS) score, there was a reduction of 0.81 units out of 10 in mean pain for patients with KOA who received prolotherapy (SMD: −0.81; 95% CI: −5.63, 4.10; p value SMD = 0.693; I2: 48.54%; pheterogeneity = 0.08).ConclusionDrawing from the data analysis performed in this meta‐analysis, it is apparent that dextrose prolotherapy exhibits promising effectiveness in reducing joint pain and stiffness, as well as improving functional performance in individuals suffering from KOA. Furthermore, it is recommended that forthcoming studies incorporate follow‐up periods to guide decisions concerning the duration of prolotherapy's effects.