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Growth of human bronchial epithelial cells at an air-liquid interface alters the response to particle exposure.

Ghio AJ, Dailey LA, Soukup JM, Stonehuerner J, Richards JH, Devlin RB - Part Fibre Toxicol (2013)

Bottom Line: Subsequently, it was not possible to attribute the observed decreases in the response of NHBE cells to differentiation alone since BEAS-2B cells, which do not differentiate, showed similar changes when grown at ALI.We conclude that growth of NHBE cells at ALI is associated with a diminished biological effect following particle exposure relative to cells submerged in media.This decreased response showed an association with increased oxygen availability.

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ABSTRACT

Background: We tested the hypothesis that normal human bronchial epithelial (NHBE) cells 1) grown submerged in media and 2) allowed to differentiate at air-liquid interface (ALI) demonstrate disparities in the response to particle exposure.

Results: Following exposure of submerged NHBE cells to ambient air pollution particle collected in Chapel Hill, NC, RNA for IL-8, IL-6, heme oxygenase 1 (HOX1) and cyclooxygenase 2 (COX2) increased. The same cells allowed to differentiate over 3, 10, and 21 days at ALI demonstrated no such changes following particle exposure. Similarly, BEAS-2B cells grown submerged in media demonstrated a significant increase in IL-8 and HOX1 RNA after exposure to NIST 1648 particle relative to the same cells exposed after growth at ALI. Subsequently, it was not possible to attribute the observed decreases in the response of NHBE cells to differentiation alone since BEAS-2B cells, which do not differentiate, showed similar changes when grown at ALI. With no exposure to particles, differentiation of NHBE cells at ALI over 3 to 21 days demonstrated significant decrements in baseline levels of RNA for the same proteins (i.e. IL-8, IL-6, HOX1, and COX2). With no exposure to particles, BEAS-2B cells grown at ALI showed comparable changes in RNA for IL-8 and HOX1. After the same particle exposure, NHBE cells grown at ALI on a transwell in 95% N2-5% CO2 and exposed to NIST 1648 particle demonstrated significantly greater changes in IL-8 and HOX1 relative to cells grown in 95% air-5% CO2.

Conclusions: We conclude that growth of NHBE cells at ALI is associated with a diminished biological effect following particle exposure relative to cells submerged in media. This decreased response showed an association with increased oxygen availability.

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RT-PCR for IL-8 (A), IL-6 (B), HOX1 (C), and COX2 (D) relative to GAPDH in unexposed NHBE cells. RNA for all proteins significantly changed with differentiation at ALI. *Significant difference relative to RNA in NHBE cells at day 0.
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Figure 3: RT-PCR for IL-8 (A), IL-6 (B), HOX1 (C), and COX2 (D) relative to GAPDH in unexposed NHBE cells. RNA for all proteins significantly changed with differentiation at ALI. *Significant difference relative to RNA in NHBE cells at day 0.

Mentions: The effect of differentiation of NHBE cells grown at ALI on baseline levels of RNA for IL-8, IL-6, HOX1, and COX2 was assessed. Quantities of mRNA for IL-8, IL-6, HOX1, and COX2 all significantly changed during differentiation of NHBE cells at ALI over 3 to 21 days (Figures 3A-3D). RNA for IL-8, IL-6, HOX1, and COX2 showed rapid changes beginning 3 days after the cells were placed at ALI. BEAS-2B cells grown at ALI also showed changes in RNA for IL-8 and HOX1 which were similar to the NHBE cells with decrements in both between 3 and 21 days of growth at ALI relative to cells grown submerged (Figures 4A and4B).


Growth of human bronchial epithelial cells at an air-liquid interface alters the response to particle exposure.

Ghio AJ, Dailey LA, Soukup JM, Stonehuerner J, Richards JH, Devlin RB - Part Fibre Toxicol (2013)

RT-PCR for IL-8 (A), IL-6 (B), HOX1 (C), and COX2 (D) relative to GAPDH in unexposed NHBE cells. RNA for all proteins significantly changed with differentiation at ALI. *Significant difference relative to RNA in NHBE cells at day 0.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750262&req=5

Figure 3: RT-PCR for IL-8 (A), IL-6 (B), HOX1 (C), and COX2 (D) relative to GAPDH in unexposed NHBE cells. RNA for all proteins significantly changed with differentiation at ALI. *Significant difference relative to RNA in NHBE cells at day 0.
Mentions: The effect of differentiation of NHBE cells grown at ALI on baseline levels of RNA for IL-8, IL-6, HOX1, and COX2 was assessed. Quantities of mRNA for IL-8, IL-6, HOX1, and COX2 all significantly changed during differentiation of NHBE cells at ALI over 3 to 21 days (Figures 3A-3D). RNA for IL-8, IL-6, HOX1, and COX2 showed rapid changes beginning 3 days after the cells were placed at ALI. BEAS-2B cells grown at ALI also showed changes in RNA for IL-8 and HOX1 which were similar to the NHBE cells with decrements in both between 3 and 21 days of growth at ALI relative to cells grown submerged (Figures 4A and4B).

Bottom Line: Subsequently, it was not possible to attribute the observed decreases in the response of NHBE cells to differentiation alone since BEAS-2B cells, which do not differentiate, showed similar changes when grown at ALI.We conclude that growth of NHBE cells at ALI is associated with a diminished biological effect following particle exposure relative to cells submerged in media.This decreased response showed an association with increased oxygen availability.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: We tested the hypothesis that normal human bronchial epithelial (NHBE) cells 1) grown submerged in media and 2) allowed to differentiate at air-liquid interface (ALI) demonstrate disparities in the response to particle exposure.

Results: Following exposure of submerged NHBE cells to ambient air pollution particle collected in Chapel Hill, NC, RNA for IL-8, IL-6, heme oxygenase 1 (HOX1) and cyclooxygenase 2 (COX2) increased. The same cells allowed to differentiate over 3, 10, and 21 days at ALI demonstrated no such changes following particle exposure. Similarly, BEAS-2B cells grown submerged in media demonstrated a significant increase in IL-8 and HOX1 RNA after exposure to NIST 1648 particle relative to the same cells exposed after growth at ALI. Subsequently, it was not possible to attribute the observed decreases in the response of NHBE cells to differentiation alone since BEAS-2B cells, which do not differentiate, showed similar changes when grown at ALI. With no exposure to particles, differentiation of NHBE cells at ALI over 3 to 21 days demonstrated significant decrements in baseline levels of RNA for the same proteins (i.e. IL-8, IL-6, HOX1, and COX2). With no exposure to particles, BEAS-2B cells grown at ALI showed comparable changes in RNA for IL-8 and HOX1. After the same particle exposure, NHBE cells grown at ALI on a transwell in 95% N2-5% CO2 and exposed to NIST 1648 particle demonstrated significantly greater changes in IL-8 and HOX1 relative to cells grown in 95% air-5% CO2.

Conclusions: We conclude that growth of NHBE cells at ALI is associated with a diminished biological effect following particle exposure relative to cells submerged in media. This decreased response showed an association with increased oxygen availability.

Show MeSH
Related in: MedlinePlus