Assessment of the biosafety profile of Galium verum in vitro on myoblasts and in ovo on chorioallantoic membrane.

BACKGROUND: Cardiovascular diseases are the first cause of death in the world. Ischemic heart disease is the main cause of heart failure. New approaches are continuously sought to identify better therapeutic success. Thereby, current research has been drawn to identifying and completing the therapeutic profile of natural sources. Galium species are representatives exhibiting diuretic and antibacterial potential in living organisms and can treat burns, wounds, and skin diseases. Moreover, it was also observed that these plants manifest cardioprotective effects as well as having antihemolytic, antioxidant, antibacterial, anti-inflammatory, immunomodulatory, and antiproliferative potential. In ischemic heart disease, Galium verum (G. verum) extract manifested preservative properties in terms of contractility, systolic and diastolic function maintenance, and reduced damage to the heart after ischemia. In addition, G. verum extract upregulated the activity of antioxidant enzymes alleviating the production of pro-oxidants.

AIM: To test the ethanolic extract of G. verum on the H9C2(2-1) cell line by evaluating the in vitro biosafety profile and in ovo irritative potential.

METHODS: Cells were tested in vitro for viability (using the MTT test), cellular morphology, cell number, confluence, nuclear morphology (by immunofluorescence staining of cell nuclei and F-actin assay) and in ovo by the hen's egg chorioallantoic membrane (CAM) test and CAM anti-irritant methods to study the irritation potential on the CAM.

RESULTS: The extract demonstrated a dose-dependent stimulatory activity. The viability increased to 170% for the dose of 55 µg/mL and decreased to 135% at 200 µg/mL. The results of cell number, confluence, and morphological analysis did not present significant changes compared with control untreated cells. The immunofluorescence assay showed insignificant apoptotic potential, and the hen's egg CAM test revealed that the extract was in the weak to moderately irritating category with an irritation score of 5.3. When applying the sample to the CAM, only slight coagulation was observed (128 s). The anti-irritant test revealed the protective potential of the extract in the vascular plexus.

CONCLUSION: The ethanolic extract of G. verum manifests a stimulating effect on cardiomyocytes, enhancing cell viability, and maintaining a normal elongated shape, cell number, and confluence, without significant signs of apoptosis and with a weak irritative effect in ovo. In addition, the extract demonstrated a protective effect against hemorrhage, lysis, and coagulation of blood vessels induced by sodium dodecyl sulfate on the CAM.