Niu Hui, Xiao Jiarui, Lou Xiaoli, Guo Lingling, Zhang Yongsheng, Yang Runhuai, Yang Hao, Wang Shouli, Niu Fuzhou
Department of Pathology, Second Affiliated Hospital of Soochow University, Suzhou, China.
School of Biology and Basic Medical Sciences, Soochow University, Suzhou, China.
Front Bioeng Biotechnol. 2022 Mar 8;10:800830. doi: 10.3389/fbioe.2022.800830. eCollection 2022.
Traditional monolayer cell cultures often fail to accurately predict the anticancer activity of drug candidates, as they do not recapitulate the natural microenvironment. Recently, three-dimensional (3D) culture systems have been increasingly applied to cancer research and drug screening. Materials with good biocompatibility are crucial to create a 3D tumor microenvironment involved in such systems. In this study, natural silk fibroin (SF) and chitosan (CS) were selected as the raw materials to fabricate 3D microscaffolds; Besides, sodium tripolyphosphate (TPP), and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) were used as cross-linking agents. The physicochemical properties of obtained scaffolds were characterized with kinds of testing methods, including emission scanning electron microscopy, x-ray photoelectron spectroscopy, fourier transform infrared spectroscopy, water absorption, and swelling ratio analysis. Cancer cell lines (LoVo and MDA-MB-231) were then seeded on scaffolds for biocompatibility examination and drug sensitivity tests. SEM results showed that EDC cross-linked scaffolds had smaller and more uniform pores with great interconnection than the TPP cross-linked scaffolds, and the EDC cross-linked scaffold exhibited a water absorption ratio around 1000% and a swelling ratio of about 72%. These spatial structures and physical properties could provide more adhesion sites and sufficient nutrients for cell growth. Moreover, both LoVo and MDA-MB-231 cells cultured on the EDC cross-linked scaffold exhibited good adhesion and spreading. CCK8 results showed that increased chemotherapeutic drug sensitivity was observed in 3D culture compared with 2D culture, particularly in the condition of low drug dose (<1 M). The proposed SF/CS microscaffold can provide a promising platform for the efficacy prediction and sensitivity screening of anticancer drugs.
传统的单层细胞培养往往无法准确预测候选药物的抗癌活性，因为它们无法重现自然微环境。近年来，三维（3D）培养系统越来越多地应用于癌症研究和药物筛选。具有良好生物相容性的材料对于构建此类系统中涉及的3D肿瘤微环境至关重要。在本研究中，选择天然丝素蛋白（SF）和壳聚糖（CS）作为原材料来制备3D微支架；此外，使用三聚磷酸钠（TPP）和1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDC）作为交联剂。通过多种测试方法对所得支架的物理化学性质进行了表征，包括发射扫描电子显微镜、X射线光电子能谱、傅里叶变换红外光谱、吸水率和溶胀率分析。然后将癌细胞系（LoVo和MDA-MB-231）接种在支架上进行生物相容性检查和药物敏感性测试。扫描电子显微镜结果表明，与TPP交联的支架相比，EDC交联的支架具有更小、更均匀且相互连通性更好的孔隙，并且EDC交联的支架表现出约1000%的吸水率和约72%的溶胀率。这些空间结构和物理性质可以为细胞生长提供更多的粘附位点和充足的营养物质。此外，在EDC交联的支架上培养的LoVo和MDA-MB-231细胞均表现出良好的粘附和铺展。CCK8结果表明，与二维培养相比，三维培养中观察到化疗药物敏感性增加，特别是在低药物剂量（<1 μM）的情况下。所提出的SF/CS微支架可为抗癌药物的疗效预测和敏感性筛选提供一个有前景的平台。
