Winter Stephen J, Miller Hunter A, Steinbach-Rankins Jill M
School of Medicine, University of Louisville School of Medicine, Louisville, KY 40202, USA.
Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY 40202, USA.
Pharmaceutics. 2021 Nov 8;13(11):1891. doi: 10.3390/pharmaceutics13111891.
A novel multicellular model composed of epithelial ovarian cancer and fibroblast cells was developed as an in vitro platform to evaluate nanovector delivery and ultimately aid the development of targeted therapies. We hypothesized that the inclusion of peptide-based scaffold (PuraMatrix) in the spheroid matrix, to represent in vivo tumor microenvironment alterations along with metastatic site conditions, would enhance spheroid cell growth and migration and alter nanovector transport. The model was evaluated by comparing the growth and migration of ovarian cancer cells exposed to stromal cell activation and tissue hypoxia. Fibroblast activation was achieved via the TGF-β1 mediated pathway and tissue hypoxia via 3D spheroids incubated in hypoxia. Surface-modified nanovector transport was assessed via fluorescence and confocal microscopy. Consistent with previous in vivo observations in ascites and at distal metastases, spheroids exposed to activated stromal microenvironment were denser, more contractile and with more migratory cells than nonactivated counterparts. The hypoxic conditions resulted in negative radial spheroid growth over 5 d compared to a radial increase in normoxia. Nanovector penetration attenuated in PuraMatrix regardless of surface modification due to a denser environment. This platform may serve to evaluate nanovector transport based on ovarian ascites and metastatic environments, and longer term, it provide a means to evaluate nanotherapeutic efficacy.
构建了一种由上皮性卵巢癌和成纤维细胞组成的新型多细胞模型,作为体外平台来评估纳米载体递送,并最终助力靶向治疗的开发。我们假设,在球体基质中加入基于肽的支架(PuraMatrix)以模拟体内肿瘤微环境变化及转移部位情况,会增强球体细胞的生长和迁移,并改变纳米载体的转运。通过比较暴露于基质细胞活化和组织缺氧条件下的卵巢癌细胞的生长和迁移情况,对该模型进行了评估。通过TGF-β1介导的途径实现成纤维细胞活化,通过在缺氧条件下培养三维球体实现组织缺氧。通过荧光和共聚焦显微镜评估表面修饰的纳米载体转运。与先前在腹水和远处转移的体内观察结果一致,与未活化的对应物相比,暴露于活化基质微环境的球体密度更高、收缩性更强且迁移细胞更多。与正常氧条件下球体半径增加相比,缺氧条件导致球体在5天内呈负向径向生长。由于环境更致密,无论表面修饰如何,纳米载体在PuraMatrix中的渗透都会减弱。该平台可用于基于卵巢腹水和转移环境评估纳米载体转运,从长远来看,它提供了一种评估纳米治疗效果的方法。