Limits...
In vitro systems toxicology approach to investigate the effects of repeated cigarette smoke exposure on human buccal and gingival organotypic epithelial tissue cultures.

Schlage WK, Iskandar AR, Kostadinova R, Xiang Y, Sewer A, Majeed S, Kuehn D, Frentzel S, Talikka M, Geertz M, Mathis C, Ivanov N, Hoeng J, Peitsch MC - Toxicol. Mech. Methods (2014)

Bottom Line: CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues.Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset.These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products.

View Article: PubMed Central - PubMed

Affiliation: Philip Morris International R&D, Philip Morris Products S.A. , Neuchâtel , Switzerland.

ABSTRACT
Smoking has been associated with diseases of the lung, pulmonary airways and oral cavity. Cytologic, genomic and transcriptomic changes in oral mucosa correlate with oral pre-neoplasia, cancer and inflammation (e.g. periodontitis). Alteration of smoking-related gene expression changes in oral epithelial cells is similar to that in bronchial and nasal epithelial cells. Using a systems toxicology approach, we have previously assessed the impact of cigarette smoke (CS) seen as perturbations of biological processes in human nasal and bronchial organotypic epithelial culture models. Here, we report our further assessment using in vitro human oral organotypic epithelium models. We exposed the buccal and gingival organotypic epithelial tissue cultures to CS at the air-liquid interface. CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues. Using microarray technology, gene-set analysis and a novel computational modeling approach leveraging causal biological network models, we identified CS impact on xenobiotic metabolism-related pathways accompanied by a more subtle alteration in inflammatory processes. Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset. These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products.

Show MeSH

Related in: MedlinePlus

Tissue viability and epithelial barrier function assessment. LDH activity was measured in the culture medium immediately after exposure (0 h), and at 4, 24 and 48 h PE of CS of the buccal (A) and gingival (B) tissue cultures. TEER was measured at 48 h PE to CS in buccal (C) and gingival (D) tissue cultures. The charts on the right show the positive control tests using Triton X-100 treatment. Means ± SEM are shown (N = 3 inserts following a single exposure run). Asterisk indicates significant p < 0.05 compared with the air-exposed control within each of the post-exposure time-point, Dunnett adjusted for multiple comparison. Abbreviations: CS, cigarette smoke; LDH, lactate dehydrogenase; PE, post-exposure; RFU, raw fluorescence unit; TEER, transepithelial electrical resistance; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4219813&req=5

f2: Tissue viability and epithelial barrier function assessment. LDH activity was measured in the culture medium immediately after exposure (0 h), and at 4, 24 and 48 h PE of CS of the buccal (A) and gingival (B) tissue cultures. TEER was measured at 48 h PE to CS in buccal (C) and gingival (D) tissue cultures. The charts on the right show the positive control tests using Triton X-100 treatment. Means ± SEM are shown (N = 3 inserts following a single exposure run). Asterisk indicates significant p < 0.05 compared with the air-exposed control within each of the post-exposure time-point, Dunnett adjusted for multiple comparison. Abbreviations: CS, cigarette smoke; LDH, lactate dehydrogenase; PE, post-exposure; RFU, raw fluorescence unit; TEER, transepithelial electrical resistance; SEM, standard error of the mean.

Mentions: Cell viability was assessed by measuring the levels of LDH released in the cultured basolateral medium at the various post-exposure time-points. In the buccal tissues, both concentrations of CS did not cause a significant increase of LDH release at all post-exposure time-points at 0, 4, 24 and 48 h (Figure 2A). In the gingival tissues (Figure 2B), one condition (19.7 % CS at 4 h post-exposure) resulted in a statistically significant increase in LDH release.Figure 2.


In vitro systems toxicology approach to investigate the effects of repeated cigarette smoke exposure on human buccal and gingival organotypic epithelial tissue cultures.

Schlage WK, Iskandar AR, Kostadinova R, Xiang Y, Sewer A, Majeed S, Kuehn D, Frentzel S, Talikka M, Geertz M, Mathis C, Ivanov N, Hoeng J, Peitsch MC - Toxicol. Mech. Methods (2014)

Tissue viability and epithelial barrier function assessment. LDH activity was measured in the culture medium immediately after exposure (0 h), and at 4, 24 and 48 h PE of CS of the buccal (A) and gingival (B) tissue cultures. TEER was measured at 48 h PE to CS in buccal (C) and gingival (D) tissue cultures. The charts on the right show the positive control tests using Triton X-100 treatment. Means ± SEM are shown (N = 3 inserts following a single exposure run). Asterisk indicates significant p < 0.05 compared with the air-exposed control within each of the post-exposure time-point, Dunnett adjusted for multiple comparison. Abbreviations: CS, cigarette smoke; LDH, lactate dehydrogenase; PE, post-exposure; RFU, raw fluorescence unit; TEER, transepithelial electrical resistance; SEM, standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219813&req=5

f2: Tissue viability and epithelial barrier function assessment. LDH activity was measured in the culture medium immediately after exposure (0 h), and at 4, 24 and 48 h PE of CS of the buccal (A) and gingival (B) tissue cultures. TEER was measured at 48 h PE to CS in buccal (C) and gingival (D) tissue cultures. The charts on the right show the positive control tests using Triton X-100 treatment. Means ± SEM are shown (N = 3 inserts following a single exposure run). Asterisk indicates significant p < 0.05 compared with the air-exposed control within each of the post-exposure time-point, Dunnett adjusted for multiple comparison. Abbreviations: CS, cigarette smoke; LDH, lactate dehydrogenase; PE, post-exposure; RFU, raw fluorescence unit; TEER, transepithelial electrical resistance; SEM, standard error of the mean.
Mentions: Cell viability was assessed by measuring the levels of LDH released in the cultured basolateral medium at the various post-exposure time-points. In the buccal tissues, both concentrations of CS did not cause a significant increase of LDH release at all post-exposure time-points at 0, 4, 24 and 48 h (Figure 2A). In the gingival tissues (Figure 2B), one condition (19.7 % CS at 4 h post-exposure) resulted in a statistically significant increase in LDH release.Figure 2.

Bottom Line: CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues.Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset.These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products.

View Article: PubMed Central - PubMed

Affiliation: Philip Morris International R&D, Philip Morris Products S.A. , Neuchâtel , Switzerland.

ABSTRACT
Smoking has been associated with diseases of the lung, pulmonary airways and oral cavity. Cytologic, genomic and transcriptomic changes in oral mucosa correlate with oral pre-neoplasia, cancer and inflammation (e.g. periodontitis). Alteration of smoking-related gene expression changes in oral epithelial cells is similar to that in bronchial and nasal epithelial cells. Using a systems toxicology approach, we have previously assessed the impact of cigarette smoke (CS) seen as perturbations of biological processes in human nasal and bronchial organotypic epithelial culture models. Here, we report our further assessment using in vitro human oral organotypic epithelium models. We exposed the buccal and gingival organotypic epithelial tissue cultures to CS at the air-liquid interface. CS exposure was associated with increased secretion of inflammatory mediators, induction of cytochrome P450s activity and overall weak toxicity in both tissues. Using microarray technology, gene-set analysis and a novel computational modeling approach leveraging causal biological network models, we identified CS impact on xenobiotic metabolism-related pathways accompanied by a more subtle alteration in inflammatory processes. Gene-set analysis further indicated that the CS-induced pathways in the in vitro buccal tissue models resembled those in the in vivo buccal biopsies of smokers from a published dataset. These findings support the translatability of systems responses from in vitro to in vivo and demonstrate the applicability of oral organotypical tissue models for an impact assessment of CS on various tissues exposed during smoking, as well as for impact assessment of reduced-risk products.

Show MeSH
Related in: MedlinePlus