Costic acid, a sesquiterpene from Nectandra barbellata (Lauraceae), attenuates sponge implant-induced inflammation, angiogenesis and collagen deposition in vivo.

Sesquiterpenes are a class of metabolites derived from plant species with immunomodulatory activity. In this study, we evaluated the effects of treatment with costic acid on inflammation, angiogenesis, and fibrosis induced by subcutaneous sponge implants in mice. One sponge disc per animal was aseptically implanted in the dorsal region of the mice and treated daily with costic acid (at concentrations of 0.1, 1, and 10 µg diluted in 10 µL of 0.5% DMSO) or 0.5% DMSO (control group). After 9 days of treatment, the animals were euthanized, and the implants collected for further analysis. Treatment with costic acid resulted in the reduction of the inflammatory parameters evaluated compared to the control group, with a decrease in the levels of inflammatory cytokines and chemokines (TNF, CXCL-1, and CCL2) and in the activity of MPO and NAG enzymes. Costic acid administration altered the process of mast cell degranulation. We also observed a reduction in angiogenic parameters, such as a decrease in the number of blood vessels, the hemoglobin content, and the levels of VEGF and FGF cytokines. Finally, when assessing implant-induced fibrogenesis, we observed a reduction in the levels of the pro-fibrogenic cytokine TGF-ß1, and lower collagen deposition. The results of this study demonstrate, for the first time, the anti-inflammatory, anti-angiogenic, and anti-fibrotic effects of costic acid in an in vivo model of chronic inflammation and reinforce the therapeutic potential of costic acid.

An In Vivo Assessment of the Effect of Hexane Extract from Endlicheria paniculata Branches and Its Main Compound, Methyldehydrodieugenol B, on Murine Sponge-Induced Inflammation.

The present study aims to explore the anti-inflammatory potential activity of the hexane extract from branches (HEB) of Endlicheria paniculata (Lauraceae) and its main compound, methyldehydrodieugenol B, in the inflammatory response induced by a murine implant sponge model. HPLC-ESI/MS analysis of HEB led to the identification of six chemically related neolignans, with methyldehydrodieugenol B as the main compound. An in silico analysis of the pharmacokinetic parameters of the identified compounds suggested moderate solubility but good absorption and biodistribution in vivo. Thus, the treatment of mice with HEB using in vivo assays indicated that HEB promoted pro-inflammatory, antiangiogenic, and antifibrogenic effects, whereas treatment with methyldehydrodieugenol B caused anti-inflammatory, antifibrogenic, and antiangiogenic effects. The obtained results shown the therapeutic potential of HEB and methyldehydrodieugenol B in the treatment of pathologies associated with inflammation and angiogenesis, including chronic wounds.

Biseugenol from Ocotea cymbarum (Lauraceae) attenuates inflammation, angiogenesis and collagen deposition of sponge-induced fibrovascular tissue in mice.

Several species of the genus Ocotea are used in traditional medicine due to their anti-inflammatory and analgesic properties. In this work we sought to investigate the effects of biseugenol, the main component of the hexane extract from the leaves of Ocotea cymbarum (Lauraceae), during a chronic inflammatory process induced by polyester-polyurethane sponge in mice. In addition to the inflammatory component, sponge discs also allowed us to evaluate parameters associated with the formation of new blood vessels and the deposition and organization of the extracellular matrix, processes that are related to the chronification of the inflammatory response. Daily treatment with biseugenol (0.1, 1 or 10 µg in 10 µl of 0.5% DMSO) inhibited the synthesis of inflammatory cytokines (TNF-α, CXCL-1 and CCL2) and the neutrophil and macrophage infiltrate into to the implants, indirectly evaluated by the activity of myeloperoxidase and N-acetyl-ß-D-glycosaminidase enzymes, respectively. In implants treated with biseugenol, we observed a reduction in angiogenesis, assessed through histological quantification of mean number of blood vessels, the levels of the pro-angiogenic cytokines FGF and VEGF and the activity of metalloproteinases. Except for VEGF levels, all mentioned parameters showed significant reductions after treatment with biseugenol. Finally, the administration of the compound also reduced TGF-ß1 levels, collagen synthesis and deposition, in addition to modifying the organization of the newly formed matrix, presenting a potential anti-fibrotic effect. Therefore, our results demonstrate the potential therapeutic use of biseugenol for the treatment of a series of pathological conditions, where parameters associated with inflammation, angiogenesis and fibrogenesis are deregulated.

α-zingiberene, a sesquiterpene from essential oil from leaves of Casearia sylvestris, suppresses inflammatory angiogenesis and stimulates collagen deposition in subcutaneous implants in mice.

α-zingiberene is a phytochemical of the sesquiterpenes class, the major constituent of the essential oil from the leaves of Casearia sylvestris, a plant widely used in traditional medicine for the treatment of inflammatory diseases, tumours, and bacterial infections. In the present study, we evaluated the effects of daily administration of α-zingiberene (0.01, 0.1 and 1 µg diluted in 10 µl of 0.5% DMSO) on the inflammatory, angiogenic, and fibrogenic components, induced by subcutaneous sponge implants in an animal model. Treatment with sesquiterpene resulted in a reduction in macrophage activation, as well as in mean blood vessels and in the activity of metalloproteinases 2 and 9. Furthermore, it resulted in an increase in collagen deposition near the implants. These results show the therapeutic potential of α-zingiberene in the treatment of pathologies, in which processes such as inflammation and angiogenesis are exacerbated, or even for the treatment of chronic wounds.

Topic use of Annona crassiflora Mart. contributes to wound healing due to the antioxidant and proliferative effects of fibroblasts.

Annona crassiflora Mart. is a species native to the Cerrado biome, whose fruit is known as araticum or marolo. Plant parts are widely used in folk medicine to treat inflammation and pain associated with rheumatism, wounds, venereal diseases, snakebites, and microbial infections. Thus, we investigated a fraction rich in phenolic compounds (PCAc) obtained from the crude extract of the peel of these fruits on non-cytotoxic, anti-inflammatory, antioxidant, and collagen biosynthesis properties in the healing of wounds induced on the back of BALB/c mice. For the control group, the induced wounds were not treated and for the others, wounds were treated topically with vehicle or vehicle plus PCAc. Both fractions contained in PCAc demonstrated effective protection on fibroblasts. We highlight the effect of the ethyl acetate fraction which, in addition to the protective effect, has a proliferative activity on these cells. In addition, PCAc caused improvement in healing after 7 days of treatment and in the longest period of treatment with PCAc (7, 14, and 21 days) there was a greater contraction of the wound, accompanied by resolution of the inflammatory process, antioxidant defense, increasing collagen synthesis, and modulation of metalloproteinases. PCAc demonstrated better re-epithelialization and organization of the dermis at the end of treatment. The changes promoted by the phenolic compounds of A. crassiflora were important in the healing process, especially in activities related to inflammation, oxidative stress, and fibrogenesis.

Wound healing activity of the hydroethanolic extract of the leaves of Maytenus ilicifolia Mart. Ex Reis.

BACKGROUND AND AIM: Maytenus ilicifolia has analgesic, healing, antioxidant and anti-inflammatory properties. This study evaluated effect of the hydroalcoholic extract of M. ilicifolia leaves on skin wound repair. EXPERIMENTAL PROCEDURE: Wounds were induced on mice and treated with the extract. The treatment was performed daily, until day 7 after wound induction. Wound closure was measured and the features of the repaired tissue were investigated, including mast cell quantification, neutrophil and macrophage activities, collagen deposition, angiogenesis, and pro-metalloproteases and metalloproteases 2 and 9 activity (pro-MMPs and MMPs). RESULTS AND CONCLUSION: The M. ilicifolia extract accelerated the closure of wounds. The extract at a concentration of 4% was found to be effective, presenting anti-inflammatory effects and hemoglobin increased, along with increased soluble, total and type III collagens in the wound. In addition, there was an increase in pro-MMP9 and MMP9 activity after day 7th of treatment. The phenolic compounds and tannins present in this plant could be associated with the anti-inflammatory and healing activities observed in this study. Therefore, the ability to modulate essential parameters for accelerated and adequate healing as shown here suggests that the use of standardised extracts of M. ilicifolia and its fractions enriched in polyphenols may represent a therapeutic strategy for the treatment of wounds.

Sesquiterpene Polygodial from Drimys brasiliensis (Winteraceae) Down-Regulates Implant-Induced Inflammation and Fibrogenesis in Mice.

Drimys brasiliensis (Winteraceae) has been investigated in traditional medicine for its anti-inflammatory properties to treat gastric ulcers and allergic and respiratory system diseases as well as for cancer treatment. In this work, we investigate the ability of the sesquiterpene polygodial, isolated from D. brasiliensis stem barks, to modulate the chronic inflammatory response induced by polyester-polyurethane sponge implants in C57BL/6J mice. Daily treatment with polygodial inhibited the macrophage content in the implants as determined by the activity of the N-acetyl-ß-d-glucosaminidase enzyme as well as decreased the levels of CXCL1/KC and CCL2/JE/MCP-1 pro-inflammatory chemokines and the presence of mast cells along the formed fibrovascular tissue. Similarly, the deposition of a new extracellular matrix (total collagen and type I and III collagen fibers) as well as the production of the TGF-ß1 cytokine were attenuated in implants treated with polygodial, showing for the first time its antifibrogenic capacity. The hemoglobin content, the number of newly formed vessels, and the levels of VEGF cytokine, which were used as parameters for the assessment of the neovascularization of the implants, did not change after treatment with polygodial. The anti-inflammatory and antifibrogenic effects of polygodial over the components of the granulation tissue induced by the sponge implant indicate a therapeutic potential for the treatment of inflammatory diseases associated with the development of fibrovascular tissue.

Dehydrodieugenol B and hexane extract from Endlicheria paniculata regulate inflammation, angiogenesis, and collagen deposition induced by a murine sponge model.

The present study reports the evaluation of hexane extract from Endlicheria paniculata and its main metabolite dehydrodieugenol B in the inflammatory response induced by a murine implant sponge model. As a result, a reduction in the inflammatory markers (myeloperoxidase and N-acetyl-ß-D-glucosaminidase) and number of mast cells were observed in comparison to the control group. All doses were also able to reduce angiogenic parameters evaluated in fibrovascular tissue. In implants treated with dehydrodieugenol B a reduction in total collagen deposition and types I and III collagen fibers were observed, while an increased in total collagen deposition and types I and III collagen fibers were observed in the treatment with hexane extract. Docking studies into cyclooxygenase-2 active site revealed that the dehydrodieugenol B had binding modes and energies comparable with celecoxib, diclofenac and ibuprofen. Therefore, dehydrodieugenol B was able to alter key components of chronic inflammation, resulting in a reduced inflammatory response and also presenting antifibrogenic and antiangiogenic effects. However, treatment with hexane extract resulted in a reduced inflammatory response with antiangiogenic effects, but caused fibrogenic effects.

Pro-Fibrogenic and Anti-Inflammatory Potential of a Polyphenol-Enriched Fraction from Annona crassiflora in Skin Repair.

A polyphenol-enriched fraction from Annona crassiflora fruit peel (PEF-Ac) containing chlorogenic acid, epi-catechin, procyanidins B2 and C1, quercetin-glucoside, kaempferol, and caffeoyl-glucoside was investigated for its anti-inflammatory, pro-angiogenic, and profibrogenic potential in the healing of cutaneous wounds. Four wounds were performed on the back of C57 mice and the lesions were treated with the vehicle (Vaseline and lanolin) and PEF-Ac at concentrations of 2%, 4%, and 6% for 4 and 7 d. Neutrophils and macrophages activities were evaluated indirectly by the activity of myeloperoxidase and N-acetyl-ß-D-glycosaminidase, angiogenesis was evaluated by hemoglobin dosing and vessel count in histological sections, and collagen deposition was assessed from histological sections stained with picrosirius red. PEF-Ac demonstrated anti-inflammatory activity, with reduced activities of neutrophil and macrophage in the cutaneous wounds. In addition, there was an increase in the synthesis of types I and III collagen, as well as in the percentage of wound closure, mainly after 4 d of treatment. On the other hand, PEF-Ac did not present an effective pro-angiogenic activity. A. crassiflora fruit peel showed anti-inflammatory and profibrogenic properties, indicating a promising natural source of bioactive molecules for treatment of cutaneous wounds.

DisBa-01 inhibits angiogenesis, inflammation and fibrogenesis of sponge-induced-fibrovascular tissue in mice.

Integrins are involved in a number of physio-pathological processes including wound healing, chronic inflammation and neoplasias. Blocking its activity is potentially of therapeutic value in these conditions. We investigated whether DisBa-01, a recombinant His-tag RGD-disintegrin from Bothrops alternatus snake venom, could modulate key events (inflammatory cell recruitment/activation, neovascularization and extracellular matrix deposition) of the proliferative fibrovascular tissue induced by polyether polyurethane sponge implants in mice. The hemoglobin content (µg/mg wet tissue), blood flow measurements (laser Doppler perfusion imaging) and number of vessels in the implants, used as indices of vascularization, showed that the disintegrin dose-dependently reduced angiogenesis in the implants relative to the Saline-treated group. DisBa-01 inhibited neutrophil and macrophage content as determined by the myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAG) activities, respectively. Similarly, down regulation of the fibrogenic component studied (collagen deposition) was observed in DisBa-01-treated implants. VEGF, bFGF, TNF-α, CXCL1 and CCL2 levels were also decreased by the disintegrin. The inhibitory effect of this αvß3-blocking disintegrin on the angiogenic, inflammatory, and fibrogenic components of the fibrovascular tissue induced by the synthetic matrix extends the range of DisBa-01 actions and may indicate its therapeutic potential in controlling angiogenesis in fibroproliferative diseases.