Microbiological control and antibacterial action of a propolis-containing mouthwash and control of dental plaque in humans

<p>Propolis is a bee product with several biological properties. This study aimed at investigating a propolis-containing mouthwash, its organoleptic properties, microbial contamination and its antibacterial action <i>in vitro</i>. This mouthwash was assessed <i>in vivo</i> to control dental plaque in humans. The presence of microorganisms was analyzed and the minimum inhibitory concentration against <i>Streptococcus mutans</i> was determined. A comparative study was done <i>in vivo</i> using propolis, chlorhexidine, and propolis plus chlorhexidine in lower concentrations for 14 days. Dental plaque was analyzed by the Patient Hygiene Performance (PHP) index. The odontological product was yellow, cloudy, free of microbial contamination, and exerted an inhibitory action <i>in vitro</i>. Individuals who used a propolis-containing mouthwash for 14 consecutive days in combination or not to chlorhexidine showed a similar PHP index to chlorhexidine alone. The product exerted an antibacterial action <i>in vitro</i> and <i>in vivo</i>, exhibiting a positive action in the control of dental plaque.</p

Nitric Oxide and Brazilian Propolis Combined Accelerates Tissue Repair by Modulating Cell Migration, Cytokine Production and Collagen Deposition in Experimental Leishmaniasis

<div><p>The fact that drugs currently used in the treatment of <i>Leishmania</i> are highly toxic and associated with acquired resistance has promoted the search for new therapies for treating American tegumentary leishmaniasis (ATL). In this study, BALB/c mice were injected in the hind paw with <i>Leishmania (Leishmania) amazonensis</i> and subsequently treated with a combination of nitric oxide (NO) donor (cis-[Ru(bpy) ₂imN(NO)](PF₆)₃) (Ru-NO), given by intraperitoneal injection, and oral Brazilian propolis for 30 days. Ru-NO reached the center of the lesion and increased the NO level in the injured hind paw without lesion exacerbation. Histological and immunological parameters of chronic inflammation showed that this combined treatment increased the efficacy of macrophages, determined by the decrease in the number of parasitized cells, leading to reduced expression of proinflammatory and tissue damage markers. In addition, these drugs in combination fostered wound healing, enhanced the number of fibroblasts, pro-healing cytokines and induced collagen synthesis at the lesion site. Overall, our findings suggest that the combination of the NO donor Ru-NO and Brazilian propolis alleviates experimental ATL lesions, highlighting a new therapeutic option that can be considered for further <i>in vivo</i> investigations as a candidate for the treatment of cutaneous leishmaniasis.</p></div

Effect of Ru-NO and propolis treatment on lesion development and parasite load of BALB/c mice infected with <i>L</i>. <i>amazonensis</i>.

<p>Animals were inoculated in the right hind paw with 1 x 10⁵ promastigote forms. <b>A)</b> After eight weeks of infection, mice were treated with saline (i.p.) (■), Glucantime (33 μmol.kg^-1 day^-1, i.p.) (□), propolis (5 mg.kg^-1 day^-1, p.o.) (▼),Ru-NO (0.385 μmol.kg^-1 day^-1, i.p.) (◆) or Ru-NO plus propolis (0.385μmol.kg^-1 day^-1, i.p. + 5 mg.kg^-1 day^-1, p.o.) (◯) for 30 days, and the lesion was measured once a week. <b>B)</b> At the end of treatment, the number of <i>Leishmania</i> kDNA was determined by real-time quantitative PCR. The results represent the mean ± SEM of lesion size for each group (n = 5). Significant difference relative to the infected control * <i>P</i><0.05 and ** <i>P</i><0.01, unpaired t-test.</p

Ru-NO plus propolis decreased protein nitration at the lesion site.

<p>Expression of <b>A)</b> iNOS and <b>B)</b> Nitrotyrosine determined by immunohistochemical assays of paw sections of infected mice at 12 weeks post-infection. Infected BALB/c mice were treated with Ru-NO (0.385 μmol.kg-1 day-1,i.p.), propolis (5 mg.kg-1 day-1, p.o.) or Glucantime (33 μmol.kg-1 day-1, i.p.) for 4 weeks post-lesion appearance. The dotted line (—) represents the uninfected control. The result was expressed as the mean ± SEM of five animals per group. Significant difference relative to infected control * P<0.05, unpaired t-test.</p

Ru-NO by intraperitoneal injection is able to reach the center of the lesion.

<p>Animals were inoculated with 1 x 10⁵<i>L</i>. <i>amazonensis</i> promastigote forms in the right hind paw, and after lesion appearance, they were treated intraperitoneally with Ru-NO (0.385 μmol.kg^-1 day^-1). <b>A)</b> EDS analysis of paw sections was performed in infected control and <b>B)</b> RuNO treated group, to identify the elements present at the lesion site.</p

Effect of Ru-NO + propolis on healing process at the lesion site.

<p>The paw sections were analyzed for <b>A)</b> number of fibroblasts (H&E), <b>B)</b> TGF-β1 by Western blotting. <b>C)</b> Photomicrographs of paw sections stained with picrosirius technique <b>D)</b> and the quantification of collagen deposition. The paw sections were analyzed at a final magnification of 200x. The data represent the mean±SEM of five animals per group. The dotted line (—) represents the uninfected control. Significant difference relative to infected control ** <i>P</i><0.01, ***<i>P</i><0.0001, unpaired t-test. Scale bars = 50 μm.</p

Effect of Ru-NO plus propolis on inflammatory process at lesion site.

<p>BALB/c mice were infected with 1x10⁵ promastigotes of <i>L</i>. <i>amazonensis</i> in the right hind paw and treated with Ru-NO (0.385 μmol.kg^-1 day^-1,i.p.), propolis (5 mg.kg^-1 day^-1, p.o.) or Glucantime (33 μmol.kg^-1 day^-1, i.p.) for 4 weeks post-lesion appearance. The paw sections were analyzed for number of <b>A)</b> macrophages (H&E), <b>B)</b> lymphocytes (H&E), <b>C)</b> CD4⁺ T (IHC), and <b>D)</b> CD8⁺ T (IHC) at a final magnification of 200x. The paw supernatants were assayed for expression of <b>E)</b> NF-κB(p65), <b>F)</b> TNF-α, <b>G)</b> IL-10 and <b>H)</b> STAT3 by Western blotting. The dotted line (—) represents the uninfected control. Bars represent the mean ± SEM (n = 5). Significant difference relative to the infected control * <i>P</i><0.05, ** <i>P</i><0.01 and ***<i>P</i> < 0.0001, unpaired t-test.</p

NO level was increased in the lesion after treatment with Ru-NO.

<p>NO/peroxynitrite was estimated by chemiluminescence in supernatant of macerated paw. The result was expressed as the mean ± SEM of five animals per group. Bars are represented by the medians of each group. (AUC: area under the curve). Significant difference relative to the infected control * <i>P</i><0.05 and ** <i>P</i><0.01, unpaired t-test.</p