Figure 9
From: Cigarette smoke extract impairs gingival epithelial barrier function
JAM1 involved in vitamin C-induced restoration of CSE-disturbed barrier function against LPS and PGN in human gingival epithelium. (a) Schematic image of culture insert system and (b) representative confocal microscopic cross-sectional images of three-dimensional culture of IHGE cells (DAPI; cyan, mouse monoclonal anti-JAM1: FITC; green). A multilayer of IHGE cells expressing shLuc or shJAM1 was cultured in the upper compartment, with vitamin C administered to samples in the upper compartments. Following 1 h of incubation, Seven Stars SCE and fluorescent tracers were administered, and the tissues were cultured for 3 h, after which culture medium from the lower compartment was analyzed using spectrometry. (c, d) Permeability of gingival epithelial tissues to FITC-P. gingivalis LPS (c) or PGN (d) expressing indicated shRNA, with or without addition of Seven Stars CSE and vitamin C. Three-dimensional tissues in culture inserts were treated with vitamin C and incubated for 1 h. Seven Stars CSE and FITC-labeled tracers were then administered to tissues in the upper compartment, and those were cultured for 3 h, after which transmission of the tracer from the upper to lower compartment was analyzed using spectrometry. Results are expressed as fold change relative to untreated shLuc-expressing cells. Values are shown as the mean ± SD of eight technical replicates. *p < 0.05, one-tailed t test (closed testing procedure).
