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Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans.

Zia Q, Khan AA, Swaleha Z, Owais M - Int J Nanomedicine (2015)

Bottom Line: Interestingly, AmB-bearing PGA nanoparticles were found to inhibit biofilm formation to a considerable extent.In summary, AmB-PGA nanoparticles showed highly attenuated toxicity when compared with Fungizone, while retaining equivalent active antifungal properties.This study indicates that the AmB-PGA preparation could be a promising treatment for various fungal infections.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, India.

ABSTRACT
In the present study, we developed a self-assembled biodegradable polyglutamic acid (PGA)-based formulation of amphotericin B (AmB) and evaluated its in vitro antifungal potential against Candida albicans. The AmB-loaded PGA nanoparticles were prepared in-house and had a mean size dimension of around 98±2 nm with a zeta potential of -35.2±7.3 mV. Spectroscopic studies revealed that the drug predominantly acquires an aggregated form inside the formulation with an aggregation ratio above 2. The PGA-based AmB formulation was shown to be highly stable in phosphate-buffered saline as well as in serum (only 10%-20% of the drug was released after 10 days). The AmB-PGA nanoparticles were less toxic to red blood cells (<15% lysis at an AmB concentration of 100 μg/mL after 24 hours) when compared with Fungizone(®), a commercial antifungal product. An MTT assay showed that the viability of mammalian cells (KB and RAW 264.7) was negligibly affected at AmB concentrations as high as 200 μg/mL. Histopathological examination of mouse kidney revealed no signs of tissue necrosis. The AmB-PGA formulation showed potent antimicrobial activity similar to that of Fungizone against C. albicans. Interestingly, AmB-bearing PGA nanoparticles were found to inhibit biofilm formation to a considerable extent. In summary, AmB-PGA nanoparticles showed highly attenuated toxicity when compared with Fungizone, while retaining equivalent active antifungal properties. This study indicates that the AmB-PGA preparation could be a promising treatment for various fungal infections.

No MeSH data available.


Related in: MedlinePlus

Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample.Notes: (A) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. (B) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation.Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone®; AmB-L, Ambisome®.
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f4-ijn-10-1769: Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample.Notes: (A) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. (B) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation.Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone®; AmB-L, Ambisome®.

Mentions: Before introducing the novel formulation of AmB–PGA as a potential antifungal therapy approach, we investigated the associated intrinsic toxicity issues both in vitro and in vivo. Given that hemolysis is a serious toxic manifestation of AmB-D,85 we tested for acute toxicity using an in vitro red blood cell toxicity test. The association of AmB with polymers resulted in a large decrease in their hemolytic activity. At a lower concentration (1 μg/mL) and shorter duration (1 hour), no significant hemolysis was observed with any of the formulations (Figure 4A). In fact, no hemolysis was observed with the PGA formulation across the whole range of tested concentrations, ie, 1–100 μg/mL (maximum leakage was 4.0%±0.9%; P<0.001). AmB-L, used as a control, also showed negligible hemolysis (2.60%±0.7%). However, the positive control, AmB-D, showed a sharp increase in toxicity as the concentration was increased from 5 μg/mL to 10 μg/mL (20.0%±5.0% and 90.0%±7.3% respectively); with complete hemolysis at the 50 μg/mL concentration of AmB (Figure 4A).


Self-assembled amphotericin B-loaded polyglutamic acid nanoparticles: preparation, characterization and in vitro potential against Candida albicans.

Zia Q, Khan AA, Swaleha Z, Owais M - Int J Nanomedicine (2015)

Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample.Notes: (A) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. (B) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation.Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone®; AmB-L, Ambisome®.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356689&req=5

f4-ijn-10-1769: Hemolytic activity of AmB and AmB–PGA nanoparticles. The extent of damage caused to red blood cells by the AmB formulation was measured as percent lysis of total erythrocytes used in the individual sample.Notes: (A) Hemolysis caused by the AmB–PGA formulation after 1 hour of incubation with human red blood cells. (B) Hemolysis after a 24-hour incubation period. AmB-D, AmB-L and pure AmB used in preparation of the complex were used as controls. Data are pooled from three different experiments. Each datum point is a mean ± standard deviation.Abbreviations: AmB, amphotericin B; DMSO, dimethyl sulfoxide; PGA, polyglutamic acid; AmB-D, Fungizone®; AmB-L, Ambisome®.
Mentions: Before introducing the novel formulation of AmB–PGA as a potential antifungal therapy approach, we investigated the associated intrinsic toxicity issues both in vitro and in vivo. Given that hemolysis is a serious toxic manifestation of AmB-D,85 we tested for acute toxicity using an in vitro red blood cell toxicity test. The association of AmB with polymers resulted in a large decrease in their hemolytic activity. At a lower concentration (1 μg/mL) and shorter duration (1 hour), no significant hemolysis was observed with any of the formulations (Figure 4A). In fact, no hemolysis was observed with the PGA formulation across the whole range of tested concentrations, ie, 1–100 μg/mL (maximum leakage was 4.0%±0.9%; P<0.001). AmB-L, used as a control, also showed negligible hemolysis (2.60%±0.7%). However, the positive control, AmB-D, showed a sharp increase in toxicity as the concentration was increased from 5 μg/mL to 10 μg/mL (20.0%±5.0% and 90.0%±7.3% respectively); with complete hemolysis at the 50 μg/mL concentration of AmB (Figure 4A).

Bottom Line: Interestingly, AmB-bearing PGA nanoparticles were found to inhibit biofilm formation to a considerable extent.In summary, AmB-PGA nanoparticles showed highly attenuated toxicity when compared with Fungizone, while retaining equivalent active antifungal properties.This study indicates that the AmB-PGA preparation could be a promising treatment for various fungal infections.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, India.

ABSTRACT
In the present study, we developed a self-assembled biodegradable polyglutamic acid (PGA)-based formulation of amphotericin B (AmB) and evaluated its in vitro antifungal potential against Candida albicans. The AmB-loaded PGA nanoparticles were prepared in-house and had a mean size dimension of around 98±2 nm with a zeta potential of -35.2±7.3 mV. Spectroscopic studies revealed that the drug predominantly acquires an aggregated form inside the formulation with an aggregation ratio above 2. The PGA-based AmB formulation was shown to be highly stable in phosphate-buffered saline as well as in serum (only 10%-20% of the drug was released after 10 days). The AmB-PGA nanoparticles were less toxic to red blood cells (<15% lysis at an AmB concentration of 100 μg/mL after 24 hours) when compared with Fungizone(®), a commercial antifungal product. An MTT assay showed that the viability of mammalian cells (KB and RAW 264.7) was negligibly affected at AmB concentrations as high as 200 μg/mL. Histopathological examination of mouse kidney revealed no signs of tissue necrosis. The AmB-PGA formulation showed potent antimicrobial activity similar to that of Fungizone against C. albicans. Interestingly, AmB-bearing PGA nanoparticles were found to inhibit biofilm formation to a considerable extent. In summary, AmB-PGA nanoparticles showed highly attenuated toxicity when compared with Fungizone, while retaining equivalent active antifungal properties. This study indicates that the AmB-PGA preparation could be a promising treatment for various fungal infections.

No MeSH data available.


Related in: MedlinePlus