Introduction: Knee osteoarthritis (OA) is a highly prevalent and debilitating condition with significant emotional and economic impacts. Current treatment options may only provide temporary pain relief and are not regenerative, thus the progression of knee OA is not deterred and total knee arthroplasty is inevitable. Injection therapies with orthobiologics possess regenerative potential and are an emerging treatment option. We present a prospective study aimed at examining patients with knee OA who had received an autologous platelet concentrate fluid (APCF) injection produced through a fluid volume reducer. Methods: This was an observational review of the results following an APCF injection in a cohort of patients at a single site. Patients were included in the study if they were diagnosed with K/L grade 2–3 knee OA and treated with an APCF knee injection. Patients were excluded if they had obtained an orthobiologic injection in the three months prior to study enrollment or if baseline data were unavailable. Knee score and function score were used to assess patients at the baseline and post-injection follow-ups. Results: Improvements for knee score were statistically significant for the follow-ups at three months, six months, one year, and three years. Function score improved, with statistically significant changes for the three month and three year follow-ups. Conclusions: Our study demonstrates that there is some utility in using APCF injection for knee OA, with improvements that may be sustained up to three years in some patients.