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

Figure 1  Physiopathological implications in ischemic heart disease.

Figure 2 In vitro cell viability assessment of the ethanolic extract of Galium verum (15, 35, 55, 100, and 200 µg/mL) in H9C2(2-1) cells after 24-h treatment. A one-way analysis of variance followed by Dunnett’s multiple comparisons post-test revealed the statistical differences between the untreated (control) group and the treated groups. ^aP < 0.0001. Gv_EtOH: Ethanolic extract of Galium verum.

Figure 3 Confluence and cell number in H9C2(2-1) cells 24 h after treatment with the ethanolic extract of Galium verum (15, 35, 55, 100, and 200 µg/mL). A: Cell confluence; B: Cell number. Analyzed by one-way analysis of variance and Dunnett’s multiple comparisons post-test. ^aP < 0.0001. Gv_EtOH: Ethanolic extract of Galium verum.

Figure 4 Morphology of H9C2(2-1) cells after 24 h of treatment with the ethanolic extract of Galium verum. The scale bars represent 100 µm.

Figure 5 Immunofluorescence of nuclei and actin filament structures in H9C2(2-1) cells following 24 h of treatment with 15 µg/mL and 200 µg/mL of ethanolic extract of G. verum. Gv_EtOH: Ethanolic extract of Galium verum.

Figure 6 Anti-irritative hen’s egg chorioallantoic membrane test. Stereomicroscopic images of the chorioallantoic membrane treated with controls [H₂O used as negative control; sodium dodecyl sulfate (SDS) 1% used as positive control) and test samples [ethanolic extract of Galium verum (Gv_EtOH) 200 μg/mL] before treatment (t0) and 5 min post-treatment (t5). Scale bars represent 200 μm.

Figure 7 Stereomicroscopic images of anti-irritant assay. The effects observed on the chorioallantoic membrane before and after the application of sodium dodecyl sulfate (SDS), which was used as a positive control, as well as the effects induced by SDS on the chorioallantoic membrane pretreated with the ethanolic extract of Galium verum (Gv_EtOH) extract at a concentration of 200 μg/mL demonstrated the cardioprotective effects of the Gv_EtOH.