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In vitro cytotoxic and genotoxic effects of Cissus verticillata and Sphagneticola trilobata used for treatment of Diabetes Mellitus in Brazilian folk medicine

Ana Paula Schmitz, Patrícia Weimer, Aline Maria Weschenfelder, Alana Witt Hansen, Rage Weidner Maluf, Rochele Cassanta Rossi et al. · 2021 · 4 citationsRead the paper

Cissus verticillata and Sphagneticola trilobata have been used in Brazilian folk medicine for Diabetes Mellitus treatment, although their pharmacological and toxicological profile has not been clearly established. Thus, the aim of this study was to evaluate the preclinical toxicity of the aqueous extracts of C. verticillata and S. trilobata. The main groups of secondary metabolites were investigated, and the species differed by the presence of coumarins in C. verticillata and by tannins in S. trilobata extracts. The highest contents of phenolic compounds and flavonoids were quantified in C. verticillata infusion with 2.594 ± 0.04 mg equivalents of gallic acid g-1 of extract and 1.301 ± 0.015 mg equivalents of catechin g-1 of extract, respectively. While the extract of S. trilobata showed minimum values of these compounds, with 0.002 ± 0.001 mg equivalents of gallic acid g-1 extract and 0.005 ± 0.0004 mg equivalents of catechin g-1 of extract, respectively. These differences implied the results of in vitro antioxidant activity evaluated using ferric reducing antioxidant power (FRAP), in which the sample of C. verticillata at 5 mg mL-1 showed a value of 122 µM ferrous sulfate equivalents (FSE), while S. trilobata showed 0.93 µM FSE at the same concentration. With respect to cytotoxic assay with murine fibroblast cell line (3T3) only S. trilobata exhibited cytotoxic effects measured by MTT and Sulforhodamine B assays, evidenced by the cell viability value of approximately 16%, in both tests after 24 and 72 hours of exposure of the cells to 5 mg mL-1 of the extract. Comparatively, at 5 mg mL-1 the C. verticillata extract showed cell viability of 142% and 95%, respectively, after 24 hours of cell exposure. On the other hand, both species showed genotoxic profiles evidenced by chromosomal aberrations by Allium cepa bioassay, observed by the higher percentage values of chromosome bridges, chromosome loss, and disturbed anaphase for all concentrations of both extracts than those of the negative control. The results support the characterization of the toxicological profile for both species and create an alert regarding the use of S. trilobata, which should be avoided.

What the full text shows

Shown in the lab
  • S. trilobata aqueous extract exhibited significant cytotoxicity in murine fibroblast cells. — ~16% cell viability vs. control (5 mg mL-1 concentration; 24 and 72 hours exposure; 3T3 cell line)
  • C. verticillata aqueous extract showed significantly higher antioxidant capacity than S. trilobata. — 122 µM FSE vs. 0.93 µM FSE (5 mg mL-1 concentration; FRAP assay)
  • C. verticillata contains significantly higher levels of phenolic compounds and flavonoids than S. trilobata. — >1000-fold higher phenolics; >260-fold higher flavonoids (Aqueous infusion extracts)
  • Both species demonstrated genotoxic potential. — Higher percentage of chromosome bridges, chromosome loss, and disturbed anaphase than negative control (All tested concentrations; Allium cepa bioassay)
  • C. verticillata contains kaempferol. — 45.8 μg mL-1 (HPLC quantification of extract)
Setup
Aqueous infusions of C. verticillata and S. trilobata; murine fibroblast cell line (3T3); Allium cepa bioassay.
How
Phytochemical screening and quantification (HPLC, TLC) were performed alongside FRAP antioxidant assays. Cytotoxicity was evaluated using MTT and Sulforhodamine B assays on 3T3 cells, and genotoxicity was assessed via Allium cepa chromosomal aberration analysis.
Limitations
In vitro study only; results may not fully translate to in vivo human toxicity. · Small number of tested flavonoid reference substances.

Summarized by Gemma 4 from the open-access full text · 10 Oct 2026. Read it