Background Knee osteoarthritis (OA) is a chronic and progressive degenerative disease. It resulted from mechanical and chemical disorders that damage the joint and the underlying bone. The management of knee OA is challenging due to poor self-regeneration of connective tissues. Surgical treatment with prolotherapy approaches was conducted to treat medial compartment knee OA. Aim To know the injection frequency to reach a 50% improvement in VAS score and WOMAC index. Methods Six patients who suffered from late-stage medial compartment knee OA underwent PFO followed by twelve sessions of intra-articular dextrose prolotherapy. The subjective pain score, visual analog scale (VAS), was assessed based on the patient subjectiveness before and after treatment. Patients marked the score from 0 to 10 cm to describe the current pain state. The functional index, the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index used to evaluate the Patient’s clinical symptoms. It ranges from 0 to 96 points consisting of three main sections: pain (total 20 points), stiffness (total: eight points), and physical function disability (total 68 points). Higher scores indicate severe symptoms and function. Results Four patients showed pain relief and functional improvement with more than 50% scores of VAS and WOMAC after the treatment. Two patients received more than twelve doses of intra-articular dextrose prolotherapy due to a lack of progress. Conclusion This study provides clinical evidence for a new treatment strategy for advanced knee OA. This combined therapy improves the patient’s daily activity function and postpones the need for total knee arthroplasty (TKA).