Pang Zeyang, Cady Nicole M, Cen Lujia, Schmidt Thomas M, He Xuesong, Li Jiahe
Department of Biomedical Engineering, College of Engineering and School of Medicine, University of Michigan, Ann Arbor, MI, USA.
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI, USA.
Int J Oral Sci. 2025 May 27;17(1):42. doi: 10.1038/s41368-025-00365-9.
Understanding microbial-host interactions in the oral cavity is essential for elucidating oral disease pathogenesis and its systemic implications. In vitro bacteria-host cell coculture models have enabled fundamental studies to characterize bacterial infection and host responses in a reductionist yet reproducible manner. However, existing in vitro coculture models fail to establish conditions that are suitable for the growth of both mammalian cells and anaerobes, thereby hindering a comprehensive understanding of their interactions. Here, we present an asymmetric gas coculture system that simulates the oral microenvironment by maintaining distinct normoxic and anaerobic conditions for gingival epithelial cells and anaerobic bacteria, respectively. Using a key oral pathobiont, Fusobacterium nucleatum, as the primary test bed, we demonstrate that the system preserves bacterial viability and supports the integrity of telomerase-immortalized gingival keratinocytes. Compared to conventional models, this system enhanced bacterial invasion, elevated intracellular bacterial loads, and elicited more robust host pro-inflammatory responses, including increased secretion of CXCL10, IL-6, and IL-8. In addition, the model enabled precise evaluation of antibiotic efficacy against intracellular pathogens. Finally, we validate the ability of the asymmetric system to support the proliferation of a more oxygen-sensitive oral pathobiont, Porphyromonas gingivalis. These results underscore the utility of this coculture platform for studying oral microbial pathogenesis and screening therapeutics, offering a physiologically relevant approach to advance oral and systemic health research.
了解口腔中的微生物与宿主的相互作用对于阐明口腔疾病的发病机制及其对全身的影响至关重要。体外细菌与宿主细胞共培养模型使基础研究能够以简化但可重复的方式表征细菌感染和宿主反应。然而，现有的体外共培养模型未能建立适合哺乳动物细胞和厌氧菌生长的条件，从而阻碍了对它们相互作用的全面理解。在此，我们提出了一种不对称气体共培养系统，该系统通过分别为牙龈上皮细胞和厌氧菌维持不同的常氧和厌氧条件来模拟口腔微环境。以关键的口腔致病共生菌具核梭杆菌作为主要测试对象，我们证明该系统可保持细菌活力并维持端粒酶永生化牙龈角质形成细胞的完整性。与传统模型相比，该系统增强了细菌侵袭，提高了细胞内细菌载量，并引发了更强有力的宿主促炎反应，包括CXCL10、IL-6和IL-8分泌增加。此外，该模型能够精确评估抗生素对细胞内病原体的疗效。最后，我们验证了不对称系统支持对氧气更敏感的口腔致病共生菌牙龈卟啉单胞菌增殖的能力。这些结果强调了这种共培养平台在研究口腔微生物发病机制和筛选治疗方法方面的实用性，为推进口腔和全身健康研究提供了一种生理相关的方法。
