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Evaluation of cytotoxicity and genotoxicity of Acacia aroma leaf extracts.

Mattana CM, Cangiano MA, Alcaráz LE, Sosa A, Escobar F, Sabini C, Sabini L, Laciar AL - ScientificWorldJournal (2014)

Bottom Line: The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1-20 mg/mL.The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2.However further studies are needed for longer periods including animal models to confirm the findings.

View Article: PubMed Central - PubMed

Affiliation: Área de Microbiología, Fac de Qca, Bqca y Fcia, UNSL, San Luis, Argentina.

ABSTRACT
Acacia aroma, native plant from San Luis, Argentina, is commonly used as antiseptic and for healing of wounds. The present study was conducted to investigate the in vitro cytotoxicity and genotoxicity of hot aqueous extract (HAE) and ethanolic extract (EE) of A. aroma. The cytotoxic activity was assayed by neutral red uptake assay on Vero cell. Cell treatment with a range from 100 to 5000 μg/mL of HAE and EE showed that 500 μg/mL and 100 μg/mL were the maximum noncytotoxic concentrations, respectively. The CC50 was 658 μg/mL for EE and 1020 μg/mL for HAE. The genotoxicity was tested by the single-cell gel electrophoresis comet assay. The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1-20 mg/mL. The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2. At concentrations below 5 mg/mL, the results of cytotoxicity and genotoxicity of aqueous and ethanolic extracts of Acacia aroma guarantee the safety at cell and genomic level. However further studies are needed for longer periods including animal models to confirm the findings.

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Related in: MedlinePlus

Morphological alterations of monolayers of Vero cells induced by A. aroma extracts, 20x. (a) Hot aqueous extract; (b) ethanolic extract; and (c) cell control.
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Related In: Results  -  Collection


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fig1: Morphological alterations of monolayers of Vero cells induced by A. aroma extracts, 20x. (a) Hot aqueous extract; (b) ethanolic extract; and (c) cell control.

Mentions: The neutral red is a weak cationic dye that readily penetrates cell membranes by nonionic diffusion and accumulates intracellularly in the lysosomes where it joins the lysosomal anionic matrix sites. Alterations in cell surface or membrane of the lysosome sensitive lead to lysosomal fragility and other changes that gradually become irreversible. Such changes caused by the action of xenobiotics result in a decreased uptake and binding of NR. Therefore, it is possible to distinguish dead, damaged, and living cells, which is the basis of this test. Cell treatment with a range from 100 to 5000 µg/mL of HAE and EE showed that 500 µg/mL and 100 µg/mL were the maximum noncytotoxic concentrations, respectively (Table 1). Figure 1 shows the morphological alterations of monolayers of Vero cells induced by cytotoxic concentrations of A. aroma hot aqueous extract (a) and ethanolic extract (b) with respect to cellular control that did not show any change (c). The cytotoxic concentration 50% (CC50) was tested by using neutral red uptake test. The results are graphically represented in Figures 2 and 3. Figure 2 shows the percentage of viability of Vero cells, incubated for 48 h in the presence of A. aroma HAE at different concentrations. In this study, it was found that the CC50 value was 1.8 mg/mL for HAE. Previous studies in our laboratory [19] showed MIC values ranging from 625 to 1250 µg/mL. On this base, for all microorganisms tested, this extract was not cytotoxic to Vero cells at bacteriostatics and bactericidal concentrations. Figure 3 shows the percentage of viability of Vero cells, incubated for 48 h in the presence of A. aroma EE and the CC50 value was 0.465 mg/mL. At 48 h after treatment with the extracts and before the addition of NR, cell monolayers were observed under light microscope. It was possible to detect some structural changes in those cell monolayers treated with high concentrations of extracts with respect to cellular control that did not show any change (Figure 1). Monolayers treated with high concentrations of extracts exhibited holes formation with retraction of cells even attached and generated round cells grouped and refractile intracytoplasmic granulations, in addition to cell detachment. The CC50 value of EE was not cytotoxic to Vero cells at bacteriostatics concentrations (MIC: 78–156 µg/mL) [19]. Our results are in agreement with those of Arias et al. [20]; they did not detect cytotoxicity in A. aroma extracts. Moreover, they have proposed this plant for pharmaceutical formulations.


Evaluation of cytotoxicity and genotoxicity of Acacia aroma leaf extracts.

Mattana CM, Cangiano MA, Alcaráz LE, Sosa A, Escobar F, Sabini C, Sabini L, Laciar AL - ScientificWorldJournal (2014)

Morphological alterations of monolayers of Vero cells induced by A. aroma extracts, 20x. (a) Hot aqueous extract; (b) ethanolic extract; and (c) cell control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4228826&req=5

fig1: Morphological alterations of monolayers of Vero cells induced by A. aroma extracts, 20x. (a) Hot aqueous extract; (b) ethanolic extract; and (c) cell control.
Mentions: The neutral red is a weak cationic dye that readily penetrates cell membranes by nonionic diffusion and accumulates intracellularly in the lysosomes where it joins the lysosomal anionic matrix sites. Alterations in cell surface or membrane of the lysosome sensitive lead to lysosomal fragility and other changes that gradually become irreversible. Such changes caused by the action of xenobiotics result in a decreased uptake and binding of NR. Therefore, it is possible to distinguish dead, damaged, and living cells, which is the basis of this test. Cell treatment with a range from 100 to 5000 µg/mL of HAE and EE showed that 500 µg/mL and 100 µg/mL were the maximum noncytotoxic concentrations, respectively (Table 1). Figure 1 shows the morphological alterations of monolayers of Vero cells induced by cytotoxic concentrations of A. aroma hot aqueous extract (a) and ethanolic extract (b) with respect to cellular control that did not show any change (c). The cytotoxic concentration 50% (CC50) was tested by using neutral red uptake test. The results are graphically represented in Figures 2 and 3. Figure 2 shows the percentage of viability of Vero cells, incubated for 48 h in the presence of A. aroma HAE at different concentrations. In this study, it was found that the CC50 value was 1.8 mg/mL for HAE. Previous studies in our laboratory [19] showed MIC values ranging from 625 to 1250 µg/mL. On this base, for all microorganisms tested, this extract was not cytotoxic to Vero cells at bacteriostatics and bactericidal concentrations. Figure 3 shows the percentage of viability of Vero cells, incubated for 48 h in the presence of A. aroma EE and the CC50 value was 0.465 mg/mL. At 48 h after treatment with the extracts and before the addition of NR, cell monolayers were observed under light microscope. It was possible to detect some structural changes in those cell monolayers treated with high concentrations of extracts with respect to cellular control that did not show any change (Figure 1). Monolayers treated with high concentrations of extracts exhibited holes formation with retraction of cells even attached and generated round cells grouped and refractile intracytoplasmic granulations, in addition to cell detachment. The CC50 value of EE was not cytotoxic to Vero cells at bacteriostatics concentrations (MIC: 78–156 µg/mL) [19]. Our results are in agreement with those of Arias et al. [20]; they did not detect cytotoxicity in A. aroma extracts. Moreover, they have proposed this plant for pharmaceutical formulations.

Bottom Line: The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1-20 mg/mL.The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2.However further studies are needed for longer periods including animal models to confirm the findings.

View Article: PubMed Central - PubMed

Affiliation: Área de Microbiología, Fac de Qca, Bqca y Fcia, UNSL, San Luis, Argentina.

ABSTRACT
Acacia aroma, native plant from San Luis, Argentina, is commonly used as antiseptic and for healing of wounds. The present study was conducted to investigate the in vitro cytotoxicity and genotoxicity of hot aqueous extract (HAE) and ethanolic extract (EE) of A. aroma. The cytotoxic activity was assayed by neutral red uptake assay on Vero cell. Cell treatment with a range from 100 to 5000 μg/mL of HAE and EE showed that 500 μg/mL and 100 μg/mL were the maximum noncytotoxic concentrations, respectively. The CC50 was 658 μg/mL for EE and 1020 μg/mL for HAE. The genotoxicity was tested by the single-cell gel electrophoresis comet assay. The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1-20 mg/mL. The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2. At concentrations below 5 mg/mL, the results of cytotoxicity and genotoxicity of aqueous and ethanolic extracts of Acacia aroma guarantee the safety at cell and genomic level. However further studies are needed for longer periods including animal models to confirm the findings.

Show MeSH
Related in: MedlinePlus