Platelet-rich plasma and ovarian quiescence: a bovine in vitro model for regeneration of the ovary

Abstract

Context Ovarian quiescence can be due to hormonal deficiency usually caused by apoptosis of granulosa cells responsible for oestrogen synthesis. Aim This study evaluated the regenerative effect of platelet rich plasma (PRP) on bovine in vitro models to understand its effect on granulosa cells. Methods Quiescent and healthy ovarian sections were cultured in the presence/absence of PRP for 72 h and, at different times (0, 24, 48 and 72 h), hematoxylin-eosin and immunohistochemical detection of Ki-67 were performed. Additionally, granulosa cells collected from healthy bovine ovaries were stressed with 100 ng/mL of lipopolysaccharide (LPS) in presence/absence of PRP and evaluated at 0, 4, 8 and 24 h for apoptosis by acridine orange and propidium iodide staining. Enzyme-linked immunosorbent assay tests were performed to evaluate oestrogen (E2) and anti-Müllerian hormone (AMH) concentrations on cultures of ovarian slices and granulosa cells. Key results In slides of quiescent ovaries treated with PRP, a marked and widespread positivity to Ki-67 was expressed by 40–60% of the follicular wall cells at 48 h of culture. Levels of E2 and AMH were significantly higher compared to untreated quiescent samples reaching the levels of healthy control samples. PRP counteracted the LPS effect and apoptosis (at 24 h, there were 93.44 ± 3.51% live cells with LPS + PRP compared to 37 ± 1.32% with LPS) and significantly increased concentrations of E2 and AMH. Conclusions PRP can stimulate granulosa cell proliferation and counteract inflammatory processes in vitro. Implications This treatment could improve the reproductive ability of quiescent females.