Gamete Research Centre, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Belgium.
Laboratory of Pharmaceutical Technology and Biopharmacy, Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, 2610 Wilrijk, Belgium.
Eur J Pharm Biopharm. 2021 Jan;158:143-155. doi: 10.1016/j.ejpb.2020.11.011. Epub 2020 Nov 25.
Polymeric nanoparticles (NPs) are produced using bio-compatible and bio-degradable materials such as PLGA (Poly(lactic-co-glycolic acid)). This technology provides a valuable tool to deliver molecules to the subcellular level with a relatively low risk of cytotoxicity. However their use in the field of reproductive biotechnology is not yet scientifically substantiated. The aim of the present study was to test if PLGA NPs can be taken-up by cumulus-enclosed oocytes as a first step towards potential oocyte-targeted applications to enhance oocyte quality and fertility. We conducted a series of experiments using bovine in vitro oocyte maturation as a model to study FITC-conjugated PLGA internalization (using laser-scanning confocal microscopy) and the effect of some important physical (particle size) and chemical (conjugation with PEG) modifications. We show evidence that PLGA NPs can be taken-up by cumulus cells and to a less extent by the enclosed oocytes regardless of the NP size. The NP transfer to the oocyte appear to be transcellular (via cumulus cells and transzonal projections) and paracellular (via zona pellucida). The PLGA NPs were detected in the vicinity of the oocyte as quick as 2 h post-exposure in a protein-free medium and did not compromise cumulus cell viability nor subsequent early embryo development or embryo quality. These results suggest that PLGA NPs may have promising applications as carriers for drug or molecule delivery targeting cumulus cells and oocytes.
聚合物纳米颗粒 (NPs) 是使用生物相容性和可生物降解的材料如 PLGA（聚 (乳酸-共-乙醇酸)）制造的。这项技术为将分子递送到亚细胞水平提供了有价值的工具，同时具有相对较低的细胞毒性风险。然而，它们在生殖生物技术领域的应用尚未得到科学证实。本研究的目的是测试 PLGA NPs 是否可以被卵丘包被的卵母细胞摄取，作为潜在的卵母细胞靶向应用的第一步，以提高卵母细胞质量和生育能力。我们进行了一系列实验，使用牛体外卵母细胞成熟作为模型，研究 FITC 缀合的 PLGA 内化（使用激光扫描共聚焦显微镜）和一些重要的物理（颗粒大小）和化学（与 PEG 缀合）修饰的影响。我们有证据表明，PLGA NPs 可以被卵丘细胞摄取，并且在一定程度上可以被包被的卵母细胞摄取，而与 NP 大小无关。NP 向卵母细胞的转移似乎是经细胞的（通过卵丘细胞和透明带突起）和细胞旁的（通过透明带）。在无蛋白培养基中，暴露后 2 小时内即可在卵母细胞附近检测到 PLGA NPs，并且不会损害卵丘细胞活力或随后的早期胚胎发育或胚胎质量。这些结果表明，PLGA NPs 可能具有作为药物或分子递送至卵丘细胞和卵母细胞的载体的应用前景。
