Investigating wound healing, tyrosinase inhibitory and antioxidant activities of the ethanol extracts of Salvia cryptantha and Salvia cyanescens using in vivo and in vitro experimental models

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Abstract

Ethnopharmacological Relevance

Salvia L. species are widely used against wounds and skin infections in Turkish folk medicine.

Aim of the study

The aim of the present study is to evaluate wound healing activity of the ethanol (EtOH) extracts of Salvia cryptantha and Salvia cyanescens.

Materials and methods

For the assessment of wound healing activity linear incision and circular excision wound models were employed on rats and mice. The wound healing effect was comparatively evaluated with the standard skin ointment Madecassol®. Inhibition of tyrosinase, a key enzyme in skin aging, was achieved using ELISA microplate reader. Antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenger effect, ferrous ion-chelating ability, and ferric-reducing antioxidant power (FRAP) tests.

Results and Conclusions

The EtOH extract of Salvia cryptantha treated groups of animals showed 56.5% contraction, whereas the reference drug Madecassol® showed 100% contraction. On the other hand, the same extract on linear incision wound model demonstrated a significant increase (33.2%) in wound tensile strength as compared to other groups. The results of histopathological examination maintained the upshot of linear incision and circular excision wound models as well.

These findings specify that Salvia cryptantha for wound healing activity can be appealed further phytochemical estimation for spotting its active components.

Graphical abstract

Salvia L. species are widely used against wounds and skin infections in Turkish folk medicine. The ethanolic extract of Salvia cryptantha treated groups of animals showed significant wound healing activity on linear incision and circular excision wound models.

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Introduction

Salvia L. is the largest genus of plants in the Lamiaceae family, with approximately 900 species including annual, biennial, or perennial herbs, along with woody subshrubs (Clebsch and Barner, 2003). The genus is distributed throughout the world with the center of diversity and origin appearing to be Central and Southwestern Asia. Several species are used to treat microbial infections, cancer, malaria, inflammation, loss of memory and to disinfect homes after sickness (Kamatou et al., 2008).

In Turkish folk medicine, some Salvia species have been recorded to be used against wounds, inflammatory and skin diseases (Tabata et al., 1994, Yeşilada et al., 1995, Honda et al., 1996). For instance; the roots of Salvia aethiopsis are boiled together with the resin of Abies bornmülleriana, butter, and beeswax, the mixture is then condensed into an ointment form, which is applied every day on cuts and other wounds for 5–10 days. The aerial parts of Salvia tomentosa Mill. are used against rheumatic pain. Salvia nemorosa is pounded and applied externally to stop bleedings and to heal wounds (Sezik et al., 1997). Infusion prepared with aerial parts of Salvia cryptantha is used for stomach disorders internally; the decoction obtained from aerial parts is used as antiseptic for wounds (Honda et al., 1996).

On the other hand, many people suffer from dark spots, hyperpigmentation, which is considered to be a skin disorder and causes psychological disturbance. Tyrosinase, which catalyzes the oxidation of monophenols, o-diphenols, and o-quinones, is known to be a key enzyme in melanin biosynthesis in plants and animals and used against skin aging (Khan et al., 2006).

Therefore, in the current study, our goal was to examine three relevant bioactivities; wound healing, tyrosinase inhibitory, and antioxidant activities of the ethanol (EtOH) extracts of Salvia cyanescens Boiss and Bal. and Salvia cryptantha Montbret and Bentham from Lamiaceae family. The extracts were tested in mice and rats for wound healing activity using in vivo linear incision and circular excision wound models. Tyrosinase inhibitory activity of the extracts was determined in ELISA microplate reader by high-throughput screening. Furthermore, antioxidant activity was accomplished in the extracts to utilize their relation to wound healing activity by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenger effect, ferrous ion-chelating ability, and ferric-reducing antioxidant power (FRAP) tests.

Section snippets

Plant materials

The samples of Salvia cyanescens and Salvia cryptantha were collected from the Beynam Forest in Ankara in May, 2005. The plants were identified by Prof. Dr. H. Duman from the Department of Biology, Faculty of Art and Science, Gazi University, Ankara, Turkey. The voucher specimens are deposited at the Herbarium of the Faculty of Pharmacy, Ankara University, Ankara, Turkey (AEF 23620 for Salvia cyanescens and AEF 23614 for Salvia cryptantha).

Preparation of the extracts

The aerial parts of the plants were dried at room

Results and discussion

In this study, an enquiry on wound healing activity of the EtOH extracts prepared from aerial parts of Salvia cyanescens and Salvia cryptantha, which have been used in the treatment of inflammatory disorders and wounds traditionally, were carried out on mice and rats by linear incision and circular excision wound models to verify the claimed traditional use of the plant on a scientific base. More to the point, histopathologic examination of the same extracts were also assessed. As shown in

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