Androgenetic alopecia (AGA) is a highly prevalent condition that poses a significant challenge to quality of life, yet the US Food and Drug Administration-approved treatments remain limited. Polynucleotides (PN) stimulate tissue regeneration by reducing inflammation, activation of growth factors and accelerating angiogenesis, which are vital for hair regrowth. This study aimed to assess the efficacy of PN in promoting hair regrowth by measuring changes in hair diameter and density in patients with AGA. This prospective study evaluated 28 patients with AGA who received four injection of DOT™ PN (DNA Fragment Optimizing Technology Polynucleotides) at four-week intervals. Hair diameter and density were quantitatively assessed using dermoscopic analysis. Treatment efficacy was also evaluated using patient and dermatologist satisfaction survey. Both hair diameter and density showed significant improvement, with hair diameter increased notably one month post-treatment and continuing to rise throughout the study. Total hair density improvements were evident from one months post-treatment onwards. Additionally, the dermatologist's assessment indicated stabilization in 60.7% of cases, while the patient assessment score indicated improvement in 82.1% of cases, with no serious side effects. This study demonstrates that PN effectively promotes hair regrowth in AGA patients. This is likely due to increased angiogenesis and anti-apoptotic effects. PN offers a promising new alternative for patients who have not responded to traditional treatments.