Estudo do potencial uso de creme à base do alcaloide Berberina para feridas cutâneas

Abstract

Skin lesions represent an interruption in the integrity of the skin that can either follow an adequate healing process or enter a period of persistent inflammation. The treatment of wounds has represented a challenge for health professionals, given the need for new therapeutic strategies that guarantee the normal course of the healing process and a curative result for the affected individual. The objective of the research was to develop and carry out the quality control of a cream based on the alkaloid Berberine for wound healing treatment. This is an experimental study that tested the substance in in vitro antimicrobial assays using the microdilution method in broth against Gram positive and negative bacteria, in vitro cell viability test using the colorimetric method of reduction of Methyltetrazolium with macrophages of the J774 lineage, activity anti-inflammatory to analyze the effect of the substance on Nitric Oxide synthesis and production of the pro-inflammatory cytokines TNF-α, IL-1β and PGE-2 in cultured RAW 264.7 macrophages stimulated with LPS and IFN-γ, in addition to developing a topical formulation with preliminary and accelerated stability tests. As for cell viability, the isolate showed macrophage proliferation at concentrations of 125μg/ml, with a percentage above 118.93%. For antibacterial activity, the isolate exhibited minimal inhibitory concentrations of: 125 μg/ml for Staphylococcus aureus, Staphylococcus epidermides and Escherichia cloacae, 250 μg/ml against Escherichia coli and 62.5 μg/ml for Pseudomonas aeruginosa and Shigella flexneri. As for the anti-inflammatory assay, berberine managed to reduce nitrite levels by 95.20% at a concentration of 500μg/mL; by halving this concentration (250μg/mL), the percentage increased to 95.40% and with the lowest concentration (125μg/mL) of the alkaloid used, a 93.16% reduction in nitrite levels of treated macrophages was observed . When evaluating the production of TNFα by macrophages activated by LPS+IFN-γ, it was found that the 3 concentrations of berberine promoted a decrease ranging from 44.29% - 49.13% with a statistically significant difference (p<0.001) to compare with the negative control. As for the production of IL-1β, berberine significantly reduced the levels of IL-1β (p<0.001) when compared to negative control and positive control. The reduction ranged from 18.18% - 27.27%, exhibiting lower percentages than the positive control for the 3 concentrations tested. As for the production of PGE-2, berberine showed a reduction ranging from 40.44% - 43.57%, values that did not show a significant difference compared to the reduction with the treatment made with dexamethasone (50.45%) ( p>0.05). As for the antiglycerin activity, within 48 hours, berberine presented a higher percentage of inhibition ranging from 87.29% - 90.04%. In the 72-hour period, berberine presented a variation of 84.97% - 89.06% in the concentrations of 125μg/ml, 500μg/ml, 1000μg/ml and 2000μg/ml, respectively, and in the 96-hour period, the inhibition of glycation by berberine ranged from 75.85% - 78.07%. The developed formulation showed formulation stability in preliminary and accelerated tests. The production of a coating capable of inhibiting the glycation of advanced products, reducing the levels of cytokines responsible for inflammatory reactions that prolong the healing process and promote the proliferation of cells that contribute to integumentary repair emerges as a positive alternative to improve skin clinical condition of people affected by chronic injuries.