Biofilm-stimulated epithelium modulates the inflammatory responses in co-cultured immune cells

Abstract

The gingival epithelium is a physical and immunological barrier to the microbiota of the oral cavity, which interact through soluble mediators with the immune cells that patrol the tissue at the gingival epithelium. We sought to develop a three-dimensional gingivae-biofilm interface model using a commercially available gingival epithelium to study the tissue inflammatory response to oral biofilms associated with “health”, “gingivitis” and “periodontitis”. These biofilms were developed by sequential addition of microorganisms to mimic the formation of supra- and sub-gingival plaque in vivo. Secondly, to mimic the interactions between gingival epithelium and immune cells in vivo, we integrated peripheral blood mononuclear cells and CD14+ monocytes into our three-dimensional model and were able to assess the inflammatory response in the immune cells cultured with and without gingival epithelium. We describe a differential inflammatory response in immune cells cultured with epithelial tissue, and more so following incubation with epithelium stimulated by “gingivitis-associated” biofilm. These results suggest that gingival epithelium-derived soluble mediators may control the inflammatory status of immune cells in vitro, and therefore targeting of the epithelial response may offer novel therapies. This multi-cellular interface model, both of microbial and host origin, offers a robust in vitro platform to investigate host-pathogens at the epithelial surface.

A nanocarrier system that potentiates the effect of miconazole within different interkingdom biofilms

“…To assess the effect of IONPs-CS-MCZ on pathogenic biofilms, three different in vitro multi-species biofilm models were tested, representative of caries, denture and gingivitis developed as previously described [4,27,28]. For all biofilm models, the bacterial and fungal cells were adjusted to 1 × 10 7 cells/mL in Todd Hewitt Broth (THB) medium supplemented with 0.01 mg/mL hemin and 2 µg/mL menadione [29].…”

“…Biofilm development for all models used followed similar protocols as described previously [4,27,28,30]. Briefly, biofilm formation consisted of inoculating Streptococcus species and C. albicans(500 µL) on the first day to promote primary colonization, followed by the addition of the remaining species on the second day.…”

“…For qPCR,1 µL of sample DNA was added to a mastermix solution containing 10 µL of SYBR GreenER TM (Thermo-Fisher, UK), 7 µL of UV-treated RNase-free water and 1 µL of 10 µM forward/reverse primers for each microbial genus or species. Table 1 displays the primers used in the study [4,27,28]. A total volume of 20 μl was added to MicroAmp fast-optical 96-well 0.1 ml reaction plates (Applied Biosystems, USA) and loaded into the StepOnePlus™ real time system (Applied Biosystems, USA).…”

Candida aurisphenotypic heterogeneity determines pathogenicityin vitro

“…Following co-culture with C. auris , epithelial tissue was carefully cut from the 0.5 cm 2 insert using a 19-gauge needle and washed three times in sterile PBS to remove non-adherent cells in a similar manner as previously described [25], as summarized in the schematic in Figure 1. Tissue was then fixed in 10% neutral-buffered formalin prior to embedding in paraffin.…”

Features of the clinical choice of antibacterial agents in chronic generalized periodontitis treatment by the results of microbiological and molecular genetic study

“…Пцр-диагностика позволяет в более короткие сроки, с меньшими трудозатратами и с большей точностью выявлять наличие генетического матери-original research Stomatology Оригинальные исследОвания Стоматология С воеобразие пародонтита как сосудисто-нервной дистрофии тканей пародонта состоит в том, что на начальных стадиях патологический процесс в тканях пародонта протекает в форме классического острого экссудативного воспаления с сочетанием явлений альтерации, экссудации и пролиферации, но при этом не происходит репарации поврежденных тканей и восстановления гомеостаза, так как процесс приобретает признаки хронического воспаления [1,2]. Среди прочего это обусловлено длительной и постоянной персистенцией пародонтопатогенной микрофлоры и развитием в последующем необратимой деструкции периодонта и альвеолярной кости, а также системными нарушениями иммунной системы с соответствующими морфологическими изменениями тканей, с изменениями когнитивной функции ЦНС [3][4][5]. Видовой состав микрофлоры варьирует в зависимости от тяжести течения процесса и от индивидуальных особенностей микробиоценоза полости рта [6,7].…”