부산대학교 도서관

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. Ovarian quiescence in women and in animals can be due to hormonal deficiency caused by apoptosis of granulosa cells responsible for hormone synthesis. In an alternative to the classical approach with hormonal treatment, regenerative medicine by platelet rich plasma could be a new treatment available. In this study, an in vitro model represented by ovarian slices and granulosa cells treated by platelet rich plasma stimulated granulosa cell proliferation and increased hormone levels. This treatment could improve the reproductive ability of quiescent females. (a) Green apoptotic granulosa cells with membrane budding and red dead granulose cell. (b) Live granulosa cells with defined margins after platelet rich plasma treatment. [ABSTRACT FROM AUTHOR]