Analysis of In Vitro Cyto- and Genotoxicity of Barbatimão Extract on Human Keratinocytes and Fibroblasts

Abstract

Barbatimão (Stryphnodendron adstringens, Mart.) is a native Brazilian species used in traditional medicine and some commercial preparations owing to its strong wound-healing activity. However, controversy regarding its use due to safety concerns over the potential genotoxic effect of this plant remains. In order to clarify this issue, the effect of hydroalcoholic extract of barbatimão in vitro on cell viability, DNA damage, and induction of apoptosis in two commercial cell lines of keratinocytes (HaCaT) and fibroblasts (HFF-1) was evaluated. Barbatimão stem bark hydroalcoholic extract (70% ethanol) was obtained and lyophilized for subsequent use in all experiments. The main bioactive molecules quantified by HPLC were gallic acid, caffeic acid, quercetin, catechin, and epigallocatechin gallate (EGCG). Barbatimão (0.024 to 1.99 mg/mL) was found to decrease cellular mortality as compared to the control group. GEMO assay, a noncellular DNA protocol that uses H2O2-exposed calf thymus DNA, revealed not only a genotoxic effect of barbatimão, but also a potential genoprotective action against H2O2-triggered DNA fragmentation. These results indicated that barbatimão at concentrations of 0.49 and 0.99 mg/mL, which are near to the levels found in commercial preparations, exerted an in vitro genoprotective effect on cells by decreasing the levels of DNA oxidation quantified by 8-hydroxy-2′-deoxyguanosine (8-OHdG) and reactive oxygen species (ROS) levels. Gene and protein apoptotic markers, quantified by qRT-PCR (BAX/Bcl-2 genes) and immunoassays (Caspases 3 and 8), respectively, also indicated a decrease in apoptotic events in comparison with control cells. Collectively, the results suggest that barbatimão could exert genoprotective and antiapoptotic effects on human keratinocytes and fibroblasts.

Effect of Stryphnodendron adstringens (Mart.) Coville extract in aerobic and anaerobic microorganisms and mammalian cells

“…The effect of the S. adstringens hydroalcoholic extract has also been evaluated for cell viability, DNA damage, and apoptosis induction in keratinocytes (HaCaT), and fibroblast (HFF-1) cell lines. The results have suggested that the extract may have a genoprotective and anti-apoptotic effects for human keratinocytes and fibroblasts (Pellenz et al, 2018).…”

“…Genotoxicity tests demonstrate effects on DNA, gene mutations, and chromosomal aberrations. Some studies have shown that S. adstringens maintains cell viability when analyzed on mammalian cell culture (de Oliveira et al, 2013;Henriques et al, 2016;Ishida et al, 2006), and has a protective effect on cellular genetic material (Costa et al, 2010;Pellenz et al, 2018).…”

In vitro and in vivo effect of novel GA‐CSNPs loaded col‐fibrin nanocomposite scaffold on diabetic wound healing

“…47 It was proven that Gallic acid effectively promoted wound healing with the activation of c-Jun N-terminal kinases (JNK), focal adhesion kinases (FAK), and extracellular signal-regulated kinases (Erk), increased fibroblast and keratinocyte proliferation as well as migration and decreased apoptosis in the keratinocyte and fibroblast cells. 48 Treatment with Gallic acid also helps to accelerate wound contraction, increases hydroxyproline content in the granulation tissues, helps to provide high tensile strength, increases collagen synthesis and deposition, and increases the vascularization process with decreased IL-6, NO, and TNF-α levels. 49 Besides increased fibroblast activity, a decrease in inflammation is the most crucial contribution of Gallic acid in wound healing.…”

Evidências sobre as propriedades do extrato de Stryphnodendron adstringens (Mart.) Coville (Barbatimão) para a prática clínica

“…Research, Society and Development, v. 10, n. 1, e3010111350, 2021 (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i1.11350 Hernandes et al, 2010, investigated the healing effect of back in wounds made in the skin of rats, after of treatment in which the existence of the stimulated proliferation of the keratinocytes (Hernandes et al, 2010). Other study evaluated the modulation of genotoxicity and apoptosis of the Barbatimão extract on DNA damage in human keratinocytes and dermal fibroblast and identified genoprotective effect on cells by decreasing the levels of DNA oxidation and reactive oxygen species (ROS) levels (Pellenz et al, 2018).…”

“…Since 2002, the use of phytotherapeutics medical has been considered a part of healthcare due to its potential for the development of new drugs according with World Health Organization (WHO) (Goes et al, 2020;Moraes et al, 2020;Pellenz et al, 2018;Santos et al, 2020) Stryphnodendron adstringens (Mart.) Coville (Barbatimão) is a native Brazilian species used in traditional medicine and some commercial preparations as natural product (De Santana, Voeks, & Funch, 2016).…”