Baek In-Hyuk, Helms Volkhard, Kim Youngjun
Environmental Safety Group, Korea Institute of Science & Technology Europe GmbH, Campus E71, 66123 Saarbrücken, Germany.
Center for Bioinformatics, Saarland University, 66123 Saarbrücken, Germany.
J Funct Biomater. 2024 Oct 24;15(11):314. doi: 10.3390/jfb15110314.
Combined 3D cell culture in vitro assays with microenvironment-mimicking systems are effective for cell-based screening tests of drug and chemical toxicity. Filamentous bacteriophages have diverse applications in material science, drug delivery, tissue engineering, energy, and biosensor development. Specifically, genetically modified bacteriophages have the potential to deliver therapeutic molecules or genes to targeted tumor tissues. The engineered bacteriophages in this study significantly enhanced endothelial cell migration and tube formation within the extracellular matrix (ECM). Compared to TGF-β1 alone and non-modified phages, the presence of TGF-β1 on the bacteriophages demonstrated superior performance as a continuous stimulant in the microenvironment, effectively promoting these angiogenic processes. Assays, including RT-qPCR, ELISA, and fluorescence microscopy, confirmed the expression of angiogenic markers such as CD31, validating the formation of 3D angiogenic structures. Our findings indicate that the TGF-β1 displayed by bacteriophages likely acted as a chemotactic factor, promoting the migration, proliferation, and tube formation of endothelial cells (ECs) within the ECM. Although direct contact between ECs and bacteriophages was not explicitly confirmed, the observed effects strongly suggest that TGF-β1-RGD bacteriophages contributed to the stimulation of angiogenic processes. The formation of angiogenic structures by ECs in the ECM was confirmed as three-dimensional and regulated by the surface treatment of microfluidic channels. These results suggest that biocompatible TGF-β1-displaying bacteriophages could continuously stimulate the microenvironment in vitro for angiogenesis models. Furthermore, we demonstrated that these functionalized bacteriophages have the potential to be utilized as versatile biomaterials in the field of biomedical engineering. Similar strategies could be applied to develop angiogenic matrices for tissue engineering in in vitro assays.
将3D细胞培养体外测定与模拟微环境系统相结合，对于基于细胞的药物和化学毒性筛选测试是有效的。丝状噬菌体在材料科学、药物递送、组织工程、能源和生物传感器开发等方面有多种应用。具体而言，基因工程噬菌体有潜力将治疗性分子或基因递送至靶向肿瘤组织。本研究中的工程噬菌体显著增强了细胞外基质（ECM）内内皮细胞的迁移和管腔形成。与单独的TGF-β1和未修饰的噬菌体相比，噬菌体上TGF-β1的存在作为微环境中的持续刺激物表现出卓越性能，有效促进了这些血管生成过程。包括RT-qPCR、ELISA和荧光显微镜在内的测定证实了血管生成标志物如CD31的表达，验证了3D血管生成结构的形成。我们的研究结果表明，噬菌体展示的TGF-β1可能作为趋化因子，促进ECM内内皮细胞（ECs）的迁移、增殖和管腔形成。尽管未明确证实ECs与噬菌体之间的直接接触，但观察到的效应强烈表明TGF-β1-RGD噬菌体有助于刺激血管生成过程。ECs在ECM中形成的血管生成结构被确认为三维的，并受微流控通道表面处理的调节。这些结果表明，具有生物相容性的展示TGF-β1的噬菌体可在体外持续刺激血管生成模型的微环境。此外，我们证明这些功能化噬菌体有潜力在生物医学工程领域用作通用生物材料。类似策略可应用于在体外测定中开发用于组织工程的血管生成基质。
