In vitro study after exposure to the aqueous extract of Piper amalago L. shows changes of morphology, proliferation, cytoskeleton and molecules of the extracellular matrix

Abstract

Piper amalago L. is a medicinal plant traditionally used as a healing agent for wounds, burns, abscesses, boils, and insect bites. The current study aimed to evaluate the possible effects of the aqueous crude extract obtained from P. amalago leaves, in different concentrations and in different incubation times, using the in vitro model of mouse fibroblasts (3T3). The extract was tested in different concentrations at the 24 h incubation time for analysis of cell viability, cytotoxicity, proliferation, cell morphology, immunostaining, adhesion and cell spreading assays, as well as to determine the hydroxyproline concentration and activity of the metalloproteinase MMP2. Morphologically, after exposure to the concentrations of 15 and 150 µg/mL, the cells maintained the morphology, yet a greater number of cells with more expansions of the cell body and larger than the control cells were observed. The treated cell culture also showed a greater number of cells, larger cells, a greater expansion of the cell body, adherent cells spread over the substrate, and a more juxtaposed, central and spherical nucleus. The treatment induced greater cell adhesion to the polymer, fibronectin, and collagen I. Biochemical results showed a significant increase in the hydroxyproline amino acid after exposure for 96 h. The extract did not induce loss of cell viability until the concentration reached 150 µg/mL, positively modulating proliferation, morphology, adhesion, degree of spreading, and organization of microfilaments. The extract also promoted a significant increase in the hydroxyproline amino acid.

A Review of Bioactive Compounds and Antioxidant Activity Properties of Piper Species

“…P. amalago can relieve chest pain, and the leaves have been used in traditional medicine in Central America to treat digestive problems and heal burns, abscesses, boils, and insect bites. Chewed leaves are applied to bleeding cuts [ 28 , 29 , 30 , 31 ].…”

Xenogenic bone grafting biomaterials do not interfere in the viability and proliferation of stem cells from human exfoliated deciduous teeth - an in vitro pilot study

“…This in vitro cell culture research followed similar designs, previously demonstrated like the classical work by Dominici (2006), and more recent assays as demonstrated by Santos et al (2021). MTT assay followed Mosmann (1983) Gomez et al (2017); cell proliferation followed Chiba et al (1998); and neutral red assay followed Chiba et al (1998), Repetto et al (2008) and Gomez et al (2017).…”

“…According to the prior results, one of the testing groups and the control group underwent a SEM protocol (Santos et al, 2021) .…”

Possible Mechanism of Dysphania ambrosioides (L.) Mosyakin & Clemants Seed Extract Suppresses the Migration and Invasion of Human Hepatocellular Carcinoma Cells SMMC‐7721

“…In this study, the microfilaments of treated cells were disorganized and the area of the cytoskeleton decreased, suggesting that CSE inhibits the migration of SMMC‐7721 cells by this mechanism. It has been also reported that some plant extracts, such as those of Swertia mussotii and Piper amalago L., also have this damaging effect on cancer cells [37–38] . Subsequently, the results of the scratch test and transwell test confirmed that CSE inhibited the cell proliferation of SMMC‐7721 cells by mediating their invasion and migration.…”

“…It has been also reported that some plant extracts, such as those of Swertia mussotii and Piper amalago L., also have this damaging effect on cancer cells. [37][38] Subsequently, the results of the scratch test and transwell test confirmed that CSE inhibited the cell proliferation of SMMC-7721 cells by mediating their invasion and migration.…”