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S-nitrosoglutathione accelerates recovery from 5-fluorouracil-induced oral mucositis.

Skeff MA, Brito GA, de Oliveira MG, Braga CM, Cavalcante MM, Baldim V, Holanda-Afonso RC, Silva-Boghossian CM, Colombo AP, Ribeiro RA, Moura-Neto V, Leitão RF - PLoS ONE (2014)

Bottom Line: The HPMC/GSNO formulations were found to provide sustained release of NO for more than 4 h at concentration-dependent rates of 14 to 80 nmol/mL/h.HPMC/GSNO administration also reversed the inhibitory effect of 5-FU on cell proliferation on day 14.Topical HPMC/GSNO accelerates mucosal recovery, reduces inflammatory parameters, speeds up re-epithelization and decreases levels of periodontopathic species in mucosal ulcers.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Morphogenesis, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Department of Morphology, School of Medicine, Federal University of Ceará, Fortaleza, CE, Brazil.

ABSTRACT

Introduction: Mucositis induced by anti-neoplastic drugs is an important, dose-limiting and costly side-effect of cancer therapy.

Aim: To evaluate the effect of the topical application of S-nitrosoglutathione (GSNO), a nitric oxide donor, on 5-fluorouracil (5-FU)-induced oral mucositis in hamsters.

Materials and methods: Oral mucositis was induced in male hamsters by two intraperitoneal administrations of 5-FU on the first and second days of the experiment (60 and 40 mg/kg, respectively) followed by mechanical trauma on the fourth day. Animals received saline, HPMC or HPMC/GSNO (0.1, 0.5 or 2.0 mM) 1 h prior to the 5-FU injection and twice a day for 10 or 14 days. Samples of cheek pouches were harvested for: histopathological analysis, TNF-α and IL-1β levels, immunohistochemical staining for iNOS, TNF-α, IL-1β, Ki67 and TGF-β RII and a TUNEL assay. The presence and levels of 39 bacterial taxa were analyzed using the Checkerboard DNA-DNA hybridization method. The profiles of NO released from the HPMC/GSNO formulations were characterized using chemiluminescence.

Results: The HPMC/GSNO formulations were found to provide sustained release of NO for more than 4 h at concentration-dependent rates of 14 to 80 nmol/mL/h. Treatment with HPMC/GSNO (0.5 mM) significantly reduced mucosal damage, inflammatory alterations and cell death associated with 5-FU-induced oral mucositis on day 14 but not on day 10. HPMC/GSNO administration also reversed the inhibitory effect of 5-FU on cell proliferation on day 14. In addition, we observed that the chemotherapy significantly increased the levels and/or prevalence of several bacterial species.

Conclusion: Topical HPMC/GSNO accelerates mucosal recovery, reduces inflammatory parameters, speeds up re-epithelization and decreases levels of periodontopathic species in mucosal ulcers.

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Related in: MedlinePlus

Primary cultures of oral fibroblasts from newborn rats incubated for 90 min in medium containing GSNO (a–d) or GSNO-free (e–h).The cells were immunostained using antibodies against smooth muscle anti-alpha-actin (c, d, g and h) or Ki67 (b, d, f and h), and nuclei were labeled with DAPI (a, d, e and h). Treatment of cells with 0.5 mM HPMC/GSNO for 90 min increased the expression of Ki67 (f and h) compared with untreated cells (b and d). Merged images are shown on the right (d and h). Scale bar = 10 µm.
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pone-0113378-g008: Primary cultures of oral fibroblasts from newborn rats incubated for 90 min in medium containing GSNO (a–d) or GSNO-free (e–h).The cells were immunostained using antibodies against smooth muscle anti-alpha-actin (c, d, g and h) or Ki67 (b, d, f and h), and nuclei were labeled with DAPI (a, d, e and h). Treatment of cells with 0.5 mM HPMC/GSNO for 90 min increased the expression of Ki67 (f and h) compared with untreated cells (b and d). Merged images are shown on the right (d and h). Scale bar = 10 µm.

Mentions: Figure 8 shows representative immunostaining for Ki67, a marker of proliferating cells, in myofibroblasts incubated for 90 min in medium containing GSNO (0.5 mM) (Figures 8e, 8f, 8g, 8h) or that was GSNO-free (untreated control; Figures 8a, 8b, 8c and 8d). An increase in the expression of Ki67 was observed in the cells incubated with GSNO (0.5 mM; Figures 8b and 8d) compared with untreated cells (Figures 8f and 8h). Figure 9 illustrates fibroblast proliferation (based on Ki67 expression) in the GSNO-treated cells and the untreated control cells after 30, 90 and 180 min of incubation. Treatment of cells with GSNO for 90 and 180 min, but not 30 min, resulted in an increase (P<0.05) in Ki67-positive cells compared with untreated control cells.


S-nitrosoglutathione accelerates recovery from 5-fluorouracil-induced oral mucositis.

Skeff MA, Brito GA, de Oliveira MG, Braga CM, Cavalcante MM, Baldim V, Holanda-Afonso RC, Silva-Boghossian CM, Colombo AP, Ribeiro RA, Moura-Neto V, Leitão RF - PLoS ONE (2014)

Primary cultures of oral fibroblasts from newborn rats incubated for 90 min in medium containing GSNO (a–d) or GSNO-free (e–h).The cells were immunostained using antibodies against smooth muscle anti-alpha-actin (c, d, g and h) or Ki67 (b, d, f and h), and nuclei were labeled with DAPI (a, d, e and h). Treatment of cells with 0.5 mM HPMC/GSNO for 90 min increased the expression of Ki67 (f and h) compared with untreated cells (b and d). Merged images are shown on the right (d and h). Scale bar = 10 µm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4257535&req=5

pone-0113378-g008: Primary cultures of oral fibroblasts from newborn rats incubated for 90 min in medium containing GSNO (a–d) or GSNO-free (e–h).The cells were immunostained using antibodies against smooth muscle anti-alpha-actin (c, d, g and h) or Ki67 (b, d, f and h), and nuclei were labeled with DAPI (a, d, e and h). Treatment of cells with 0.5 mM HPMC/GSNO for 90 min increased the expression of Ki67 (f and h) compared with untreated cells (b and d). Merged images are shown on the right (d and h). Scale bar = 10 µm.
Mentions: Figure 8 shows representative immunostaining for Ki67, a marker of proliferating cells, in myofibroblasts incubated for 90 min in medium containing GSNO (0.5 mM) (Figures 8e, 8f, 8g, 8h) or that was GSNO-free (untreated control; Figures 8a, 8b, 8c and 8d). An increase in the expression of Ki67 was observed in the cells incubated with GSNO (0.5 mM; Figures 8b and 8d) compared with untreated cells (Figures 8f and 8h). Figure 9 illustrates fibroblast proliferation (based on Ki67 expression) in the GSNO-treated cells and the untreated control cells after 30, 90 and 180 min of incubation. Treatment of cells with GSNO for 90 and 180 min, but not 30 min, resulted in an increase (P<0.05) in Ki67-positive cells compared with untreated control cells.

Bottom Line: The HPMC/GSNO formulations were found to provide sustained release of NO for more than 4 h at concentration-dependent rates of 14 to 80 nmol/mL/h.HPMC/GSNO administration also reversed the inhibitory effect of 5-FU on cell proliferation on day 14.Topical HPMC/GSNO accelerates mucosal recovery, reduces inflammatory parameters, speeds up re-epithelization and decreases levels of periodontopathic species in mucosal ulcers.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell Morphogenesis, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Department of Morphology, School of Medicine, Federal University of Ceará, Fortaleza, CE, Brazil.

ABSTRACT

Introduction: Mucositis induced by anti-neoplastic drugs is an important, dose-limiting and costly side-effect of cancer therapy.

Aim: To evaluate the effect of the topical application of S-nitrosoglutathione (GSNO), a nitric oxide donor, on 5-fluorouracil (5-FU)-induced oral mucositis in hamsters.

Materials and methods: Oral mucositis was induced in male hamsters by two intraperitoneal administrations of 5-FU on the first and second days of the experiment (60 and 40 mg/kg, respectively) followed by mechanical trauma on the fourth day. Animals received saline, HPMC or HPMC/GSNO (0.1, 0.5 or 2.0 mM) 1 h prior to the 5-FU injection and twice a day for 10 or 14 days. Samples of cheek pouches were harvested for: histopathological analysis, TNF-α and IL-1β levels, immunohistochemical staining for iNOS, TNF-α, IL-1β, Ki67 and TGF-β RII and a TUNEL assay. The presence and levels of 39 bacterial taxa were analyzed using the Checkerboard DNA-DNA hybridization method. The profiles of NO released from the HPMC/GSNO formulations were characterized using chemiluminescence.

Results: The HPMC/GSNO formulations were found to provide sustained release of NO for more than 4 h at concentration-dependent rates of 14 to 80 nmol/mL/h. Treatment with HPMC/GSNO (0.5 mM) significantly reduced mucosal damage, inflammatory alterations and cell death associated with 5-FU-induced oral mucositis on day 14 but not on day 10. HPMC/GSNO administration also reversed the inhibitory effect of 5-FU on cell proliferation on day 14. In addition, we observed that the chemotherapy significantly increased the levels and/or prevalence of several bacterial species.

Conclusion: Topical HPMC/GSNO accelerates mucosal recovery, reduces inflammatory parameters, speeds up re-epithelization and decreases levels of periodontopathic species in mucosal ulcers.

Show MeSH
Related in: MedlinePlus