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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

Cell death and proliferation in the cheek pouches of hamsters subjected to 5-FU-induced oral mucositis, on day 14.Oral mucositis was induced in hamsters by intraperitoneal (i.p.) injection of 5-FU followed by mechanical trauma (MT) of the cheek pouch. Animals received topical applications of a gel containing 0.5 mM S-nitrosoglutathione (GSNO) 30 min prior to 5-FU and twice daily thereafter for 10 days or 14 days. Control groups comprised healthy animals (H) and animals subjected to 5-FU-induced oral mucositis that received local applications of saline (saline). The TUNEL- and Ki67-positive cells were counted (10 fields per slide, 400×) for statistical comparisons. Bars denote the means ± standard errors of stained cells from at four slides per group (4 animals per group). *denotes a significant difference (P<0.05) compared with the Healthy group; **denotes a significant difference (P<0.05) compared with the MT group, +denotes a significant difference (P<0.05) compared with the Saline group; #denotes a significant difference (P<0.05) compared with the HPMC group. Data were analyzed using the Kruskal Wallis and Mann Whitney tests.
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pone-0113378-g006: Cell death and proliferation in the cheek pouches of hamsters subjected to 5-FU-induced oral mucositis, on day 14.Oral mucositis was induced in hamsters by intraperitoneal (i.p.) injection of 5-FU followed by mechanical trauma (MT) of the cheek pouch. Animals received topical applications of a gel containing 0.5 mM S-nitrosoglutathione (GSNO) 30 min prior to 5-FU and twice daily thereafter for 10 days or 14 days. Control groups comprised healthy animals (H) and animals subjected to 5-FU-induced oral mucositis that received local applications of saline (saline). The TUNEL- and Ki67-positive cells were counted (10 fields per slide, 400×) for statistical comparisons. Bars denote the means ± standard errors of stained cells from at four slides per group (4 animals per group). *denotes a significant difference (P<0.05) compared with the Healthy group; **denotes a significant difference (P<0.05) compared with the MT group, +denotes a significant difference (P<0.05) compared with the Saline group; #denotes a significant difference (P<0.05) compared with the HPMC group. Data were analyzed using the Kruskal Wallis and Mann Whitney tests.

Mentions: Figure 4 illustrates immunostaining for iNOS, IL-1β, TNF-α and TGF-β RII in cheek pouches of hamsters subjected to 5-FU-induced oral mucositis (Figures 3c, 3g, 3k and 3o, respectively) on the 14th day of treatment compared with the weak staining observed in the Healthy control group (Figures. 3b, 3f, 3j, 3n). Local application of 0.5 mM HPMC/GSNO for 14 days caused a considerable reduction in both iNOS and TNF-α immunostaining (Figures 3d and 3i, respectively) but had no effect on IL-1β immunostaining (Figure 4h) when compared with the untreated group comprising animals subjected to experimental mucositis and that received saline (Figures 3c and 3k, respectively). In contrast, 0.5 mM HPMC/GSNO increased TGF-β RII immunostaining in cheek pouch tissue markedly (Figure 4p) compared with the Saline group (Figure 4o). When the antibodies were replaced with PBS/BSA 5%, no immunostaining was detected (negative control; Figures 3a, 3e, 3i and 3m). Figure 5 shows the quantification of cells in cheek pouches of hamsters subjected to 5-FU-induced oral mucositis that received daily applications of saline (Saline) or 0.5 mM HPMC/GSNO for 14 days that were positive for TNF-α (Figure 5A), IL-1β (Figure 5B), iNOS (Figure 6C) or TGF-β RII (Figure 5D). A significantly greater number of TNF-α-, IL-1β- and iNOS-positive cells (P<0.05) were observed in cheek pouches of animals subjected to oral mucositis and that received saline or HPMC when compared with the Healthy group (Figures 4A, 4B and 4C, respectively). The topical application of 0.5 mM HPMC/GSNO reduced (P<0.05) the number of iNOS- and TNF-α-positive cells when compared with both the HPMC and Saline groups; however, the treatment did not reduce the number of IL-1β-positive cells (Figure 5B). Experimental oral mucositis significantly (P<0.05) increased TGF-β RII -positive cells in cheek pouch tissue compared with the Healthy group. There was also a 2-fold increase (P<0.05) in the number of TGF-β receptor-II-positive cells in the 0.5 mM HPMC/GSNO group compared with the HPMC and Saline groups.


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)

Cell death and proliferation in the cheek pouches of hamsters subjected to 5-FU-induced oral mucositis, on day 14.Oral mucositis was induced in hamsters by intraperitoneal (i.p.) injection of 5-FU followed by mechanical trauma (MT) of the cheek pouch. Animals received topical applications of a gel containing 0.5 mM S-nitrosoglutathione (GSNO) 30 min prior to 5-FU and twice daily thereafter for 10 days or 14 days. Control groups comprised healthy animals (H) and animals subjected to 5-FU-induced oral mucositis that received local applications of saline (saline). The TUNEL- and Ki67-positive cells were counted (10 fields per slide, 400×) for statistical comparisons. Bars denote the means ± standard errors of stained cells from at four slides per group (4 animals per group). *denotes a significant difference (P<0.05) compared with the Healthy group; **denotes a significant difference (P<0.05) compared with the MT group, +denotes a significant difference (P<0.05) compared with the Saline group; #denotes a significant difference (P<0.05) compared with the HPMC group. Data were analyzed using the Kruskal Wallis and Mann Whitney tests.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0113378-g006: Cell death and proliferation in the cheek pouches of hamsters subjected to 5-FU-induced oral mucositis, on day 14.Oral mucositis was induced in hamsters by intraperitoneal (i.p.) injection of 5-FU followed by mechanical trauma (MT) of the cheek pouch. Animals received topical applications of a gel containing 0.5 mM S-nitrosoglutathione (GSNO) 30 min prior to 5-FU and twice daily thereafter for 10 days or 14 days. Control groups comprised healthy animals (H) and animals subjected to 5-FU-induced oral mucositis that received local applications of saline (saline). The TUNEL- and Ki67-positive cells were counted (10 fields per slide, 400×) for statistical comparisons. Bars denote the means ± standard errors of stained cells from at four slides per group (4 animals per group). *denotes a significant difference (P<0.05) compared with the Healthy group; **denotes a significant difference (P<0.05) compared with the MT group, +denotes a significant difference (P<0.05) compared with the Saline group; #denotes a significant difference (P<0.05) compared with the HPMC group. Data were analyzed using the Kruskal Wallis and Mann Whitney tests.
Mentions: Figure 4 illustrates immunostaining for iNOS, IL-1β, TNF-α and TGF-β RII in cheek pouches of hamsters subjected to 5-FU-induced oral mucositis (Figures 3c, 3g, 3k and 3o, respectively) on the 14th day of treatment compared with the weak staining observed in the Healthy control group (Figures. 3b, 3f, 3j, 3n). Local application of 0.5 mM HPMC/GSNO for 14 days caused a considerable reduction in both iNOS and TNF-α immunostaining (Figures 3d and 3i, respectively) but had no effect on IL-1β immunostaining (Figure 4h) when compared with the untreated group comprising animals subjected to experimental mucositis and that received saline (Figures 3c and 3k, respectively). In contrast, 0.5 mM HPMC/GSNO increased TGF-β RII immunostaining in cheek pouch tissue markedly (Figure 4p) compared with the Saline group (Figure 4o). When the antibodies were replaced with PBS/BSA 5%, no immunostaining was detected (negative control; Figures 3a, 3e, 3i and 3m). Figure 5 shows the quantification of cells in cheek pouches of hamsters subjected to 5-FU-induced oral mucositis that received daily applications of saline (Saline) or 0.5 mM HPMC/GSNO for 14 days that were positive for TNF-α (Figure 5A), IL-1β (Figure 5B), iNOS (Figure 6C) or TGF-β RII (Figure 5D). A significantly greater number of TNF-α-, IL-1β- and iNOS-positive cells (P<0.05) were observed in cheek pouches of animals subjected to oral mucositis and that received saline or HPMC when compared with the Healthy group (Figures 4A, 4B and 4C, respectively). The topical application of 0.5 mM HPMC/GSNO reduced (P<0.05) the number of iNOS- and TNF-α-positive cells when compared with both the HPMC and Saline groups; however, the treatment did not reduce the number of IL-1β-positive cells (Figure 5B). Experimental oral mucositis significantly (P<0.05) increased TGF-β RII -positive cells in cheek pouch tissue compared with the Healthy group. There was also a 2-fold increase (P<0.05) in the number of TGF-β receptor-II-positive cells in the 0.5 mM HPMC/GSNO group compared with the HPMC and Saline groups.

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