Department of Bioengineering, The University of Texas at Dallas, 800 W. Campbell Road, Richardson, Texas 75080, United States.
Texas A&M University School of Dentistry, 3302 Gaston Avenue, Dallas, Texas 75246, United States.
ACS Biomater Sci Eng. 2024 Oct 14;10(10):6218-6229. doi: 10.1021/acsbiomaterials.4c01075. Epub 2024 Sep 23.
Coculture models are limited by bacteria rapidly outcompeting host mammalian cells for nutrients , resulting in mammalian cell death. The goal of this study was to develop a coculture model enabling survival of mammalian cells and oral bacterial species to assess their competition for growth on dental implant materials. Two early colonizing oral bacterial species, or , were grown in coculture with primary human macrophages or human gingival fibroblasts for up to 7 days on tissue-culture treated polystyrene or polished titanium and zirconia disks. Chloramphenicol was supplemented in cell culture medium at bacteriostatic concentrations to maintain stable bacterial inoculum size. Planktonic and adherent bacterial growth was assessed via spot plating while mammalian cell growth and attachment were evaluated using colorimetric metabolic assay and confocal fluorescence microscopy, respectively. Macrophages and fibroblasts proliferated in the presence of and maintained viability above 70% during coculture for up to 7 days on tissue-culture treated polystyrene and polished titanium and zirconia. In contrast, both mammalian cell types exhibited decreasing proliferation and surface coverage on titanium and zirconia over time in coculture with versus control. and were maintained within an order of magnitude of seeding inoculum sizes throughout coculture. Cell culture medium supplemented with antibiotics at bacteriostatic concentrations can suppress bacterial overgrowth and facilitate mammalian cell viability in coculture model systems. Within the study's limitations, oral bacteria and mammalian cell growth in coculture are comparable on polished titanium and zirconia surfaces.
共培养模型受到细菌的限制,细菌会迅速与宿主哺乳动物细胞争夺营养,导致哺乳动物细胞死亡。本研究的目的是开发一种共培养模型,使哺乳动物细胞和口腔细菌能够存活下来,以评估它们在牙种植体材料上的生长竞争。两种早期定植的口腔细菌,或,与人原代巨噬细胞或人牙龈成纤维细胞在经组织培养处理的聚苯乙烯或抛光钛和氧化锆盘上共培养长达 7 天。在细胞培养基中补充氯霉素以抑菌浓度维持稳定的细菌接种物大小。通过点平板评估浮游和附着细菌的生长,同时使用比色代谢测定法和共聚焦荧光显微镜分别评估哺乳动物细胞的生长和附着。巨噬细胞和成纤维细胞在 和 的存在下增殖,并在组织培养处理的聚苯乙烯和抛光钛和氧化锆上共培养长达 7 天,其活力保持在 70%以上。相比之下,两种哺乳动物细胞类型在与 共培养时,其增殖和表面覆盖率随时间推移在钛和氧化锆上逐渐减少,而与对照相比。和在共培养过程中始终保持在接种物大小的数量级内。在抑菌浓度下补充抗生素的细胞培养基可以抑制细菌过度生长,并促进共培养模型系统中哺乳动物细胞的活力。在研究的限制范围内,口腔细菌和哺乳动物细胞在抛光钛和氧化锆表面的共培养中的生长是可比的。