Evaluation of Wound Healing Activity of Salvia haenkei Hydroalcoholic Aerial Part Extract on in vitro and in vivo Experimental Models

Mario Scrima,^1,* Carmela Melito,^1,* Filomena Merola,¹ Antonio Iorio,¹ Nicoletta Vito,¹ Andrea Maria Giori,² Angela Ferravante<sup>1

1</sup>R&D Department, IBSA Farmaceutici Italia, Ariano Irpino, Italy; ²R&D Department, IBSA Farmaceutici Italia, Lodi, Italy

*These authors contributed equally to this work

Correspondence: Angela Ferravante
R&D Department, IBSA Farmaceutici Italia, Via Camporeale, Ariano Irpino 83031, AV, Italy
Tel +39 0825881847
Fax +39 0825881812
Email angela.ferravante@ibsa.it

Purpose: The aim of the present study was to evaluate the potential wound healing activity of the hydroalcoholic extract of Salvia haenkei on in vitro and in vivo experimental models.
Materials and Methods: Preliminary analytical characterization of the hydroalcoholic extract of Salvia haenkei was made by reversed-phase high performance liquid chromatography (RP-HPLC) that permitted identification of a qualitative fingerprint of the extract of aerial parts. The wound healing activity of the hydroalcoholic extract of Salvia haenkei was evaluated in vitro by the scratch assay on human dermal fibroblasts and human epidermal keratinocytes and in vivo by standardized mouse excisional splinting model. Real-time PCR (RT-PCR) experiments were performed to analyze gene expression levels of inflammatory markers.
Results: The scratch assay tests showed that the treatment with the hydroalcoholic extract of Salvia haenkei did not induce an increase in the fibroblasts migration rate with respect to the positive control. Instead, the hydroalcoholic extract of Salvia haenkei was effective in improving the wound closure rate on keratinocyte cell cultures with an almost total invasion of the scratch after 48 h of treatment; whereas the positive control, at the same time point, showed only a 67% reduction of the wound size. In vivo experiments showed that the groups treated with the extract of Salvia haenkei completely re-epithelized the wound in 2.7 days, a timing that was comparable with the action of the positive control that took only 2.1 days. Gene expression analysis showed that Salvia haenkei positively regulated the signaling pathway of the nuclear factor-κB (NF-κB) transcription factor.
Conclusion: The results suggested that the hydroalcoholic extract of Salvia haenkei induced a clear wound healing effect.

Keywords: wound healing, cosmetics, animal model, scratch assay