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Updated: Aug 25, 2025
A Novel In Vitro Wound Healing Assay to Evaluate Cell Migration
Published on: March 17, 2018
In Silico, In Vitro, and In Vivo Wound Healing Activity of Astragalus microcephalus Willd
Fatemeh Akbari1, Mohammad Azadbakht1, Ali Bagheri2
1Department of Pharmacognosy, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran.
Methods:
The methanolic root extract was prepared by maceration, and flavonoids were evaluated by LC/MS. In silico examination was performed based on the LC/MS results, and the binding affinity of these compounds to estrogen receptors (ERs) α and β was evaluated. Wound healing evaluation in both in vitro (NHDF cell line, by 500 μg/ml concentration of the extract, 24 h) and in vivo (Wistar rat, topical daily treated with 1.5% of the extract ointment, 21 days) conditions in comparison to control groups was conducted. Rats' control groups included silver sulfadiazine, Vaseline, and the nontreated groups.
Results:
Eleven flavonoids were detected using LC/MS. The in silico study showed that formononetin, kaempferol-based structures, quercetin-3-O-neohesperidoside, and calycosin-7-O-beta-D-glucoside had a high affinity (<-6.3) to ERs α and β. Wound closing measurement showed significant improvement in the group treated with the extract in both in vitro and in vivo assays compared to the control groups. Histopathological results confirmed these findings; inflammation factors decreased, and fibroblast proliferation, fibrosis, and epithelization increased, especially in the extract group.
Conclusion:
This study shows that Astragalus microcephalus has wound healing activity in vitro and in vivo with low toxicity due to the presence of flavonoids, especially isoflavonoids, which show a high affinity to bind to ERs α and β in the skin tissue.
Insights
Astragalus microcephalus root extract demonstrates significant wound healing capabilities in vitro and in vivo. Its flavonoid content shows high affinity for estrogen receptors, contributing to its therapeutic efficacy and low toxicity.
Area of Science:
- Pharmacology
- Natural Products Chemistry
- Dermatology
Background:
- Flavonoids are plant-derived compounds with diverse biological activities.
- Estrogen receptors (ERs) play a role in skin health and wound repair.
- Herbal extracts are explored for their therapeutic potential in wound healing.
Purpose of the Study:
- To investigate the wound healing activity of Astragalus microcephalus root extract.
- To identify key flavonoids in the extract and evaluate their binding affinity to estrogen receptors alpha (ERα) and beta (ERβ).
- To assess the extract's efficacy in both in vitro and in vivo wound models.
Main Methods:
- Methanolic root extract of Astragalus microcephalus prepared by maceration.
- Flavonoid profiling using Liquid Chromatography-Mass Spectrometry (LC/MS).
- In silico molecular docking to assess binding affinity to ERα and ERβ.
- In vitro assays using NHDF cell line and in vivo studies on Wistar rats for wound healing evaluation.
Main Results:
- Eleven flavonoids were identified, with formononetin, kaempferol derivatives, quercetin-3-O-neohesperidoside, and calycosin-7-O-beta-D-glucoside showing high affinity for ERα and ERβ.
- Significant wound closure was observed in both in vitro and in vivo models treated with the extract compared to controls.
- Histopathological analysis revealed reduced inflammation and increased fibroblast proliferation, fibrosis, and epithelization in the extract-treated groups.
Conclusions:
- Astragalus microcephalus extract possesses significant in vitro and in vivo wound healing properties.
- The observed activity is attributed to its flavonoid content, particularly isoflavonoids, which exhibit high binding affinity to ERα and ERβ.
- The extract demonstrates low toxicity, making it a promising candidate for wound care applications.
