Achyrocline satureioides (Lam.) DC (Asteraceae) Extract-Loaded Nanoemulsions as a Promising Topical Wound Healing Delivery System: In Vitro Assessments in Human Keratinocytes (HaCaT) and HET-CAM Irritant Potential.

Lucélia Albarello Balestrin,Letícia Scherer Koester,Helder Ferreira Teixeira,Elizandra Braganhol,Tainá Kreutz,Valquiria Linck Bassani,Juliana Bidone,Flávia Nathiely Silveira Fachel,Nicolly Espindola Gelsleichter,Cristiana Lima Dora

doi:10.3390/pharmaceutics13081241

Abstract

Achyrocline satureioides (Lam.) DC Asteraceae extracts (ASEs) have been investigated for the treatment of various skin disorders. This study reports the effects of ASE-loaded nanoemulsions (NEASE) on the cellular viability, death by necrosis, and migration of immortalized human keratinocytes (HaCaT cell line), as well as the irritant potential through the hen’s egg chorioallantoic membrane test (HET-CAM). NEASE exhibited a polydispersity index above 0.12, with a droplet size of 300 nm, ζ-potential of −40 mV, and content of flavonoids close to 1 mg/mL. No cytotoxicity of the ASE was observed on HaCaT by MTT assay (up to 10 µg/mL). A significant increase of HaCaT viability was observed to NEASE (up to 5 μg/mL of flavonoids), compared to treatment with the ASE. The necrosis death evaluation demonstrated that only NEASE did not lead to cell death at all the tested concentrations. The scratch assay demonstrated that NEASE was able to increase the cell migration at low flavonoid concentrations. Finally, the HET-CAM test proved the non-irritative potential of NEASE. Overall, the results indicate the potential of the proposed formulations for topical use in wound healing, in view of their promising effects on proliferation and migration in keratinocytes, combined with an indication of the absence of cytotoxicity and non-irritating potential.

Highlights

An Asteraceae extracts (ASEs)-loaded nanoemulsion was prepared in order to investigate the in vitro cytotoxicity profile, proliferative effect, and migration ability in immortalized human keratinocytes (HaCaT cell line), as well as the HET-chorioallantoic membrane (CAM) irritant potential
The results showed an increase in cell viability when ASE was incorporated in nanoemulsions (NEASE )
The evaluation of propidium iodide (PI) incorporation indicated that there was no cell death due to necrosis when treated with Nanoemulsions containing ASE (NEASE) and suggested that the incorporation of the extract into the nanoemulsion may decrease the ASE toxicity induced by necrosis and increase its proliferation ability

Summary

Introduction

A considerable number of medicines that are based on bioactive compounds from medicinal plants are available on the pharmaceutical market Such medicines contain drugs isolated from medicinal plant extracts or synthesized from compounds identified from these plant extracts [1]. Despite the difficulties in relating the pharmacological activity with a specific compound or class of compounds and in elucidating the mechanism of action involved, phytomedicines containing crude plant extracts are widely used in clinical practice [2]. These products contain a variety of compounds that can interact with 4.0/).

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Results

Conclusion