Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts

• Published in: Journal of ethnopharmacology Vol. 126; no. 3; pp. 463 - 467
• Main Authors: Fronza, M., Heinzmann, B., Hamburger, M., Laufer, S., Merfort, I.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 10-12-2009; Amsterdam; New York: Elsevier; Elsevier
• Subjects: 3T3 Cells; Animals; bioassays; Biological and medical sciences; Calendula; Calendula - chemistry; Calendula officinalis; cell migration; Cell Movement - drug effects; cell proliferation; Cell Proliferation - drug effects; chemical concentration; cultured cells; dosage; dose response; epithelial cells; Faradiol esters; faradiol myristate; Fibroblasts; Fibroblasts - drug effects; General pharmacology; Hypericum perforatum; in vitro studies; laboratory techniques; Matricaria recutita; Medical sciences; medicinal plants; medicinal properties; Mice; myristic acid; new methods; palmitates; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; plant extracts; Plant Extracts - pharmacology; tissue repair; Wound healing; Wound Healing - drug effects; Hypericum perforatum; Fibroblasts; Faradiol esters; Calendula officinalis; Wound healing; Matricaria recutita; Weed; Pharmacognosy; Essential oil plant; Guttiferae; Extract; Compositae; Wound; Medicinal plant; Ester; Healing agent; Dicotyledones; Angiospermae; Plant origin; Spermatophyta; Chamomile; Fibroblast; Quantitative analysis
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2009.09.014

Presentation of the scratch assay as a convenient and inexpensive
in vitro tool to gain first insights in the wound healing potential of plant extracts and natural compounds.
The present study deals with the optimization of the scratch assay which can be used as an
in vitro model for quantification of fibroblast migration to and proliferation into the wounded area. It is suitable for the first evaluation of the wound re-epithelialization potential of crude herbal extracts, isolated compounds and pharmaceutical preparations. As a proof of concept three preparations from traditional medicinal plants were investigated.
Swiss 3T3 albino mouse fibroblasts were used in monolayers and platelet derived growth factor as positive control. Hexane and ethanolic extracts from
Calendula officinalis and
Matricaria recutita,
Hypericum oil as well as the triterpenoids faradiol myristate and palmitate were studied. To differentiate between proliferation and migration antimitotic mitomycin C was added.
Both extracts of
Calendula officinalis stimulated proliferation and migration of fibroblasts at low concentrations, e.g. 10
μg/ml enhanced cell numbers by 64.35% and 70.53%, respectively. Inhibition of proliferation showed that this effect is mainly due to stimulation of migration. Faradiol myristate and palmitate gave comparable stimulation rates at an almost 50
μg/ml concentration, indicating that they contribute partially, but not most significantly to the wound healing effects of
Calendula preparations. Extracts from
Matricaria recutita were only moderately active.
Hypericum oil was cytotoxic at concentrations higher than 0.5
μg/ml.
The scratch assay in the present form can be used as a promising scientific approach and platform to differentiate between plant extracts known for their wound healing and their anti-inflammatory properties.