Objective: Many postmenopausal women suffering from vulvovaginal atrophy are looking for non-hormonal treatments. Platelet-rich plasma (PRP) therapy has emerged as a novel and promising approach for gynecological applications. PRP is an autologous blood product rich in growth factors used to stimulate tissue regeneration. On the other hand, hyaluronic acid (HA) is used as a treatment for vaginal dryness as it improves tissue hydration thanks to its strong capacity to retain water. This study examines the in vitro effects of PRP alone or combined with HA on vaginal fibroblasts (VFs) isolated from mucosal samples of postmenopausal women undergoing surgery for vaginal prolapse.

Methodology: Vaginal and blood samples were collected from postmenopausal women undergoing pelvic organ prolapse surgery. PRP was prepared alone or combined with HA (PRP-HA) from peripheral blood. Vaginal fibroblasts were isolated via enzymatic dissociation and cultured in 2D and 3D (spheroids) systems. Biological activities were assessed by measuring cell growth, proliferation, senescence, metabolic activity, and collagen accumulation, along with immunocytochemistry for (myo-)fibroblast markers.

Results: VFs cultured with PRP or PRP-HA showed dose-dependent higher proliferation compared with the control condition, with increased S and G2M cell cycle phases correlating with enhanced proliferation. Expression of vimentin, a protein that plays a key role in maintaining cellular structure and function, was stable, while alpha-SMA decreased, indicating a shift from myofibroblasts to fibroblasts. Collagen production, crucial for wound healing and tissue regeneration, increased under PRP or PRP-HA treatment. PRP and PRP-HA also prevented cell senescence in long-term low-density cultures. These findings were consistent across 2D and 3D culture systems.

Conclusions: This study provides in vitro evidence supporting the potential of PRP and PRP-HA as autologous treatments for vaginal rejuvenation.