Effects of Aqueous Leaf Extract of Ageratum conyzoides Linn on Keratinocytes and Fibroblasts Behavior

Background and Purpose: Healing is a natural process that repairs injured tissue. Several medicinal plants, including Ageratum conyzoides Linn., have been tested for their wound-healing properties. Because the predominant cell populations in mammalian skin are fibroblasts and keratinocytes, our study aimed to evaluate the cell-activating and anti-inflammatory properties of the aqueous leaf extract of A. conyzoides. Specifically, we sought to show that the extract promoted fibroblast and keratinocyte proliferation. Methods: Quantification of the major class of phytochemicals was done after aqueous extraction of powdered dried leaves. Keratinocytes and fibroblasts were extracted from the foreskin of newborn to two-year-old children, and cell viability was evaluated using the trypan blue test; proliferation tests, including MTT, CFSE, and PCNA measurement, were performed. Toxicity of the extract was studied using propidium iodide (PI) + annexin V assay. Results: The extract contained 82.73 ± 0.76 mg GAE/g tannins, 164.13 ± 6.565 mg QE/g flavonoids, and 94.75 ± 1.13 mg GAE/g total phenols. In the presence of the extract, a higher percentage of dead cells (30.5% at 500 mg/mL) was observed; among them, more cells were apoptotic than necrotic. The percentage of dead cells decreased as the extract concentration decreased. Without the extract, cells were viable but did not proliferate; with the extract, cells proliferated as the extract concentration increased. Fibroblast proliferation increased slightly (15.4%), whereas keratinocyte proliferation increased markedly (64.7%). Conclusion: This study shows that the aqueous leaf extract of A. conyzoides dose-dependently activated fibroblast and keratinocyte proliferation but induced moderate toxicity at the highest concentration.