Lange-Consiglio Anna, Gaspari Giulia, Riccaboni Pietro, Canesi Simone, Bosi Giampaolo, Vigo Daniele, Cremonesi Fausto
Department of Veterinary Medicine and Animal Science (DIVAS), Università degli Studi di Milano, Via dell'Università 6, Lodi (Lo) 26900, Italy.
Reprod Fertil Dev. 2023 Apr;35(7):433-444. doi: 10.1071/RD23017.
Ovarian quiescence can be due to hormonal deficiency usually caused by apoptosis of granulosa cells responsible for oestrogen synthesis.
This study evaluated the regenerative effect of platelet rich plasma (PRP) on bovine in vitro models to understand its effect on granulosa cells.
Quiescent and healthy ovarian sections were cultured in the presence/absence of PRP for 72h and, at different times (0, 24, 48 and 72h), hematoxylin-eosin and immunohistochemical detection of Ki-67 were performed. Additionally, granulosa cells collected from healthy bovine ovaries were stressed with 100ng/mL of lipopolysaccharide (LPS) in presence/absence of PRP and evaluated at 0, 4, 8 and 24h 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.
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 48h 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 24h, 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.
PRP can stimulate granulosa cell proliferation and counteract inflammatory processes in vitro .
This treatment could improve the reproductive ability of quiescent females.
卵巢静止可能是由于激素缺乏，通常是由负责雌激素合成的颗粒细胞凋亡所致。
本研究评估富血小板血浆（PRP）对牛体外模型的再生作用，以了解其对颗粒细胞的影响。
将静止且健康的卵巢切片在有/无PRP的情况下培养72小时，并在不同时间点（0、24、48和72小时）进行苏木精-伊红染色和Ki-67免疫组化检测。此外，从健康牛卵巢收集的颗粒细胞在有/无PRP的情况下用100ng/mL脂多糖（LPS）处理，并在0、4、8和24小时通过吖啶橙和碘化丙啶染色评估细胞凋亡情况。进行酶联免疫吸附测定试验以评估卵巢切片和颗粒细胞培养物中的雌激素（E2）和抗苗勒管激素（AMH）浓度。
在用PRP处理的静止卵巢切片中，培养48小时时，40%-60%的卵泡壁细胞对Ki-67呈明显且广泛的阳性表达。与未处理的静止样本相比，E2和AMH水平显著升高，达到健康对照样本的水平。PRP抵消了LPS的作用和细胞凋亡（24小时时，LPS+PRP组有93.44±3.51%的活细胞，而LPS组为37±1.32%），并显著提高了E2和AMH的浓度。
PRP可在体外刺激颗粒细胞增殖并抵消炎症过程。
这种治疗方法可能会提高静止状态雌性动物的繁殖能力。
