Limits...
Dose-response met-RANTES treatment of experimental periodontitis: a narrow edge between the disease severity attenuation and infection control.

Repeke CE, Ferreira SB, Vieira AE, Silveira EM, Avila-Campos MJ, da Silva JS, Santos CF, Campanelli AP, Trombone AP, Garlet GP - PLoS ONE (2011)

Bottom Line: At 0.5 mg up to 5 mg doses, a strong reduction in the alveolar bone loss and inflammatory cell migration were observed.Interestingly, 5 mg dose treatment resulted in the maximum inhibition of inflammatory cell migration, but resulted in a similar inhibition of bone loss when compared with the lower doses, and also resulted in increased bacterial load and CRP response.When 0.5 and 5 mg therapy regimens were compared it was observed that both therapeutic protocols were able to downregulate the levels of pro-inflammatory, Th1-type and osteoclastogenic cytokines, and CD3+ and F4/80+ cells migration to periodontal tissues, but the high dose modulates host response in a more pronounced and unspecific and excessive way, interfering also with the production of antimicrobial mediators such as MPO, iNOS and IgG, and with GR1+ and CD19+ cells migration.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, School of Dentistry of Bauru, São Paulo University-FOB/USP, Bauru, Sao Paulo, Brazil.

ABSTRACT
Chemokines and chemokine receptors have been implicated in the selective migration of leukocyte subsets to periodontal tissues, which consequently influences the disease outcome. Among these chemoattractants, the chemokines CCL3, CCL4 and CCL5 and its receptors, CCR1 and CCR5, have been associated with increased disease severity in mice and humans. Therefore, in this study we investigated the modulation of experimental periodontitis outcome by the treatment with a specific antagonist of CCR1 and 5 receptors, called met-RANTES. C57Bl/6 mice was orally infected with Aggregatibacter actinomycetemcomitans and treated with 0.05, 0.1, 0.5, 1.5 and 5 mg doses of met-RANTES on alternate days, and evaluated by morphometric, cellular, enzymatic and molecular methods. At 0.5 mg up to 5 mg doses, a strong reduction in the alveolar bone loss and inflammatory cell migration were observed. Interestingly, 5 mg dose treatment resulted in the maximum inhibition of inflammatory cell migration, but resulted in a similar inhibition of bone loss when compared with the lower doses, and also resulted in increased bacterial load and CRP response. When 0.5 and 5 mg therapy regimens were compared it was observed that both therapeutic protocols were able to downregulate the levels of pro-inflammatory, Th1-type and osteoclastogenic cytokines, and CD3+ and F4/80+ cells migration to periodontal tissues, but the high dose modulates host response in a more pronounced and unspecific and excessive way, interfering also with the production of antimicrobial mediators such as MPO, iNOS and IgG, and with GR1+ and CD19+ cells migration. Our results demonstrate a thin line between beneficial immunoregulation and impaired host defense during experimental periodontitis, and the determination of the exact equilibrium point is mandatory for the improvement of immune-targeted therapy of periodontitis.

Show MeSH

Related in: MedlinePlus

The effect of met-RANTES treatment at different doses in the modulation of alveolar bone loss, inflammatory cell influx and bacterial load in experimental periodontal disease.C57Bl/6 mice were infected orally with A. actinomycetemcomitans and treated with met-RANTES at 0.05, 0.1, 0.5, 1.5 and 5 mg doses or (V) veicule, (-) non treated and (C) control non-infected mice and evaluated at 30 day post-infection for: A) alveolar bone loss quantification, performed through the measurements of cement-enamel junction-alveolar bone crest (CEJ-ABC) area in the palatal face of maxillary molars; B) total leukocyte counts of inflammatory infiltrate, performed in a Neubauer chamber; and C) A. actinomycetemcomintans load (AA DNA) in periodontal tissues, quantified by Real-TimePCR, using SybrGreen System and the cycle threshold (Ct) method; all performed as described in the Material and methods; *p<0.05 versus non-treated mice, unpaired t-test. Different italic low case letters represent statistically significant differences among the doses in the same experimental group (P<0.05; One-way ANOVA).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3140528&req=5

pone-0022526-g001: The effect of met-RANTES treatment at different doses in the modulation of alveolar bone loss, inflammatory cell influx and bacterial load in experimental periodontal disease.C57Bl/6 mice were infected orally with A. actinomycetemcomitans and treated with met-RANTES at 0.05, 0.1, 0.5, 1.5 and 5 mg doses or (V) veicule, (-) non treated and (C) control non-infected mice and evaluated at 30 day post-infection for: A) alveolar bone loss quantification, performed through the measurements of cement-enamel junction-alveolar bone crest (CEJ-ABC) area in the palatal face of maxillary molars; B) total leukocyte counts of inflammatory infiltrate, performed in a Neubauer chamber; and C) A. actinomycetemcomintans load (AA DNA) in periodontal tissues, quantified by Real-TimePCR, using SybrGreen System and the cycle threshold (Ct) method; all performed as described in the Material and methods; *p<0.05 versus non-treated mice, unpaired t-test. Different italic low case letters represent statistically significant differences among the doses in the same experimental group (P<0.05; One-way ANOVA).

Mentions: Our first aim was to determine the dose of met-RANTES which result in maximum decrease of experimental PD severity parameters (Fig. 1). When compared to untreated and vehicle-treated mice, met-RANTES doses of 0.1, 0.5, 1.5 and 5 mg were found to significantly inhibit both alveolar bone loss and inflammatory cell influx. When the distinct doses were compared, the 0.1 mg doses was found to be less effective than 0.5, 1.5 and 5 mg in the attenuation of bone loss while no significant differences were found between these three higher doses. Regarding inflammatory cell counts, our results demonstrate that as a general rule the dose increase resulted in a gradual decrease of inflammatory cell influx into periodontal tissues, except for non-significant differences between the doses of 0.5 and 1.5, and 1.5 and 5 mg. When the bacterial load was evaluated, our results demonstrated that only the 5 mg dose interfered in the control of infection, as revealed by the significant increase in AA DNA levels in periodontal tissues when compared to untreated, vehicle-treated and to the other met-RANTES doses groups. During the experimental period the mice mortality was absent and no significant change in mice weight among the experimental groups was verified (data not shown). Our results demonstrate that 10X dose range resulted in distinct modulation of experimental PD outcome from the tissue destruction and infectious viewpoints, and therefore the doses of 0.5 and 5 mg were selected for the subsequent evaluations.


Dose-response met-RANTES treatment of experimental periodontitis: a narrow edge between the disease severity attenuation and infection control.

Repeke CE, Ferreira SB, Vieira AE, Silveira EM, Avila-Campos MJ, da Silva JS, Santos CF, Campanelli AP, Trombone AP, Garlet GP - PLoS ONE (2011)

The effect of met-RANTES treatment at different doses in the modulation of alveolar bone loss, inflammatory cell influx and bacterial load in experimental periodontal disease.C57Bl/6 mice were infected orally with A. actinomycetemcomitans and treated with met-RANTES at 0.05, 0.1, 0.5, 1.5 and 5 mg doses or (V) veicule, (-) non treated and (C) control non-infected mice and evaluated at 30 day post-infection for: A) alveolar bone loss quantification, performed through the measurements of cement-enamel junction-alveolar bone crest (CEJ-ABC) area in the palatal face of maxillary molars; B) total leukocyte counts of inflammatory infiltrate, performed in a Neubauer chamber; and C) A. actinomycetemcomintans load (AA DNA) in periodontal tissues, quantified by Real-TimePCR, using SybrGreen System and the cycle threshold (Ct) method; all performed as described in the Material and methods; *p<0.05 versus non-treated mice, unpaired t-test. Different italic low case letters represent statistically significant differences among the doses in the same experimental group (P<0.05; One-way ANOVA).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3140528&req=5

pone-0022526-g001: The effect of met-RANTES treatment at different doses in the modulation of alveolar bone loss, inflammatory cell influx and bacterial load in experimental periodontal disease.C57Bl/6 mice were infected orally with A. actinomycetemcomitans and treated with met-RANTES at 0.05, 0.1, 0.5, 1.5 and 5 mg doses or (V) veicule, (-) non treated and (C) control non-infected mice and evaluated at 30 day post-infection for: A) alveolar bone loss quantification, performed through the measurements of cement-enamel junction-alveolar bone crest (CEJ-ABC) area in the palatal face of maxillary molars; B) total leukocyte counts of inflammatory infiltrate, performed in a Neubauer chamber; and C) A. actinomycetemcomintans load (AA DNA) in periodontal tissues, quantified by Real-TimePCR, using SybrGreen System and the cycle threshold (Ct) method; all performed as described in the Material and methods; *p<0.05 versus non-treated mice, unpaired t-test. Different italic low case letters represent statistically significant differences among the doses in the same experimental group (P<0.05; One-way ANOVA).
Mentions: Our first aim was to determine the dose of met-RANTES which result in maximum decrease of experimental PD severity parameters (Fig. 1). When compared to untreated and vehicle-treated mice, met-RANTES doses of 0.1, 0.5, 1.5 and 5 mg were found to significantly inhibit both alveolar bone loss and inflammatory cell influx. When the distinct doses were compared, the 0.1 mg doses was found to be less effective than 0.5, 1.5 and 5 mg in the attenuation of bone loss while no significant differences were found between these three higher doses. Regarding inflammatory cell counts, our results demonstrate that as a general rule the dose increase resulted in a gradual decrease of inflammatory cell influx into periodontal tissues, except for non-significant differences between the doses of 0.5 and 1.5, and 1.5 and 5 mg. When the bacterial load was evaluated, our results demonstrated that only the 5 mg dose interfered in the control of infection, as revealed by the significant increase in AA DNA levels in periodontal tissues when compared to untreated, vehicle-treated and to the other met-RANTES doses groups. During the experimental period the mice mortality was absent and no significant change in mice weight among the experimental groups was verified (data not shown). Our results demonstrate that 10X dose range resulted in distinct modulation of experimental PD outcome from the tissue destruction and infectious viewpoints, and therefore the doses of 0.5 and 5 mg were selected for the subsequent evaluations.

Bottom Line: At 0.5 mg up to 5 mg doses, a strong reduction in the alveolar bone loss and inflammatory cell migration were observed.Interestingly, 5 mg dose treatment resulted in the maximum inhibition of inflammatory cell migration, but resulted in a similar inhibition of bone loss when compared with the lower doses, and also resulted in increased bacterial load and CRP response.When 0.5 and 5 mg therapy regimens were compared it was observed that both therapeutic protocols were able to downregulate the levels of pro-inflammatory, Th1-type and osteoclastogenic cytokines, and CD3+ and F4/80+ cells migration to periodontal tissues, but the high dose modulates host response in a more pronounced and unspecific and excessive way, interfering also with the production of antimicrobial mediators such as MPO, iNOS and IgG, and with GR1+ and CD19+ cells migration.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, School of Dentistry of Bauru, São Paulo University-FOB/USP, Bauru, Sao Paulo, Brazil.

ABSTRACT
Chemokines and chemokine receptors have been implicated in the selective migration of leukocyte subsets to periodontal tissues, which consequently influences the disease outcome. Among these chemoattractants, the chemokines CCL3, CCL4 and CCL5 and its receptors, CCR1 and CCR5, have been associated with increased disease severity in mice and humans. Therefore, in this study we investigated the modulation of experimental periodontitis outcome by the treatment with a specific antagonist of CCR1 and 5 receptors, called met-RANTES. C57Bl/6 mice was orally infected with Aggregatibacter actinomycetemcomitans and treated with 0.05, 0.1, 0.5, 1.5 and 5 mg doses of met-RANTES on alternate days, and evaluated by morphometric, cellular, enzymatic and molecular methods. At 0.5 mg up to 5 mg doses, a strong reduction in the alveolar bone loss and inflammatory cell migration were observed. Interestingly, 5 mg dose treatment resulted in the maximum inhibition of inflammatory cell migration, but resulted in a similar inhibition of bone loss when compared with the lower doses, and also resulted in increased bacterial load and CRP response. When 0.5 and 5 mg therapy regimens were compared it was observed that both therapeutic protocols were able to downregulate the levels of pro-inflammatory, Th1-type and osteoclastogenic cytokines, and CD3+ and F4/80+ cells migration to periodontal tissues, but the high dose modulates host response in a more pronounced and unspecific and excessive way, interfering also with the production of antimicrobial mediators such as MPO, iNOS and IgG, and with GR1+ and CD19+ cells migration. Our results demonstrate a thin line between beneficial immunoregulation and impaired host defense during experimental periodontitis, and the determination of the exact equilibrium point is mandatory for the improvement of immune-targeted therapy of periodontitis.

Show MeSH
Related in: MedlinePlus