1 School of Engineering, Institute of Life Technologies, University of Applied Sciences , HES-SO Valais/Wallis, Sion, Switzerland .
2 R&D, Pharmalp SA , Conthey, Switzerland .
Tissue Eng Part C Methods. 2018 Jul;24(7):412-417. doi: 10.1089/ten.TEC.2018.0042.
Candida albicans frequently causes recurrent intimal infectious disease (ID). This demands the treatment of multiple phases of the infection. The objective of this study was to uncover the host-pathogen interaction using two-dimensional (2D) epithelium cell-barrier and three-dimensional (3D) subepithelium tissue cells of human mucosa. The 2D cell cultures assessed C. albicans adhesion. Addition of the antifungal drug Fluconazol did not inhibit the adhesion, despite its pathogen growth inhibition (minimal inhibitory concentration value 0.08 μg/mL). A 3D tissue was engineered in multitranswells by placing human fibroblast cultures on a thick porous scaffold. This contained the yeast placed in the top compartment and prevented passive penetration. After 28 h, the pathogen transmigrated the barrier and was collected in the bottom compartment. A change in pathogen morphology was observed where hypha formed and grew to be 231 μm long after 28 h. The hypha was thus long enough to cross the 200 μm thick 3D tissue. The 3D infection was inhibited by addition of Fluconazol (0.08 μg/mL), confirming that penetration is dependent on pathogen growth. In conclusion, ID was reconstituted step-by-step on 2D epithelium surface and in 3D connective tissue of human mucosa. Fluconazol growth-inhibition of the pathogen C. albicans was confirmed in the 3D tissue. We thus propose that this ID in vitro test is suitable for the identification and characterization of new treatments against C. albicans.
白色念珠菌常引起复发性内膜感染性疾病（ID）。这需要对感染的多个阶段进行治疗。本研究的目的是使用二维（2D）上皮细胞屏障和三维（3D）人黏膜下组织细胞来揭示宿主-病原体相互作用。2D 细胞培养评估了 C. albicans 的黏附性。尽管抗真菌药物氟康唑（氟康唑）抑制了病原体的生长（最小抑制浓度值为 0.08μg/mL），但其并没有抑制黏附。在多 Transwell 中通过将人成纤维细胞培养物置于厚多孔支架上来构建 3D 组织。这将酵母放在顶部隔室中，防止被动渗透。28 小时后，病原体穿过屏障并被收集在底部隔室中。观察到病原体形态发生变化，其中菌丝形成并在 28 小时后长到 231μm。因此，菌丝足够长可以穿过 200μm 厚的 3D 组织。加入氟康唑（0.08μg/mL）抑制了 3D 感染，证实穿透依赖于病原体的生长。总之，ID 逐步在 2D 上皮表面和人黏膜的 3D 结缔组织中重建。在 3D 组织中证实了抗真菌药物氟康唑对病原体 C. albicans 的生长抑制作用。因此，我们提出这种体外 ID 试验适合鉴定和表征针对 C. albicans 的新治疗方法。
