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Gold Nanoparticle-Photosensitizer Conjugate Based Photodynamic Inactivation of Biofilm Producing Cells: Potential for Treatment of C. albicans Infection in BALB/c Mice.

Sherwani MA, Tufail S, Khan AA, Owais M - PLoS ONE (2015)

Bottom Line: Photodynamic therapy (PDT) has been found to be effective in inhibiting biofilm producing organisms.We investigated the photodynamic effect of gold nanoparticle (GNP) conjugated photosensitizers against Candida albicans biofilm.The GNP-PS conjugate based PDT was found to effectively kill both C. albicans planktonic cells and biofilm populating hyphal forms.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, 202002, India.

ABSTRACT

Background: Photodynamic therapy (PDT) has been found to be effective in inhibiting biofilm producing organisms. We investigated the photodynamic effect of gold nanoparticle (GNP) conjugated photosensitizers against Candida albicans biofilm. We also examined the photodynamic efficacy of photosensitizer (PS) conjugated GNPs (GNP-PS) to treat skin and oral C. albicans infection in BALB/c mice.

Methods: The biomimetically synthesized GNPs were conjugated to photosensitizers viz. methylene blue (MB) or toluidine blue O (TB). The conjugation of PSs with GNPs was characterized by spectroscopic and microscopic techniques. The efficacy of gold nanoparticle conjugates against C. albicans biofilm was demonstrated by XTT assay and microscopic studies. The therapeutic efficacy of the combination of the GNP conjugates against cutaneous C. albicans infection was examined in mouse model by enumerating residual fungal burden and histopathological studies.

Results: The GNP-PS conjugate based PDT was found to effectively kill both C. albicans planktonic cells and biofilm populating hyphal forms. The mixture of GNPs conjugated to two different PSs significantly depleted the hyphal C. albicans burden against superficial skin and oral C. albicans infection in mice.

Conclusion: The GNP-PS conjugate combination exhibits synergism in photodynamic inactivation of C. albicans. The GNP conjugate based PDT can be employed effectively in treatment of cutaneous C. albicans infections in model animals. The antibiofilm potential of PDT therapy can also be exploited in depletion of C. albicans on medical appliances such as implants and catheters etc.

No MeSH data available.


Related in: MedlinePlus

Characterization of in-house synthesized GNPs and PS conjugation onto their surface.(A) Characterization of in-house synthesized gold nanoparticles. (a) Ultraviolet-visible- near infrared spectra of gold nanoparticles synthesized by exposing various amounts of Aloe vera extract to a fixed volume of HAuCl4 solution (10−3 M), keeping the final volume of reaction mixture 10 ml that was made up for each sample by increment in Aloe vera extract content for 24 h. Absorbance is seen to be increasing exhibiting two major peaks, a larger one at 540 nm, the characteristic peak of GNPs. (b) Representative TEM image of gold nanoparticles synthesized using Aloe vera leaf extract exhibiting a large population of spherical gold nanoparticles of approx. 10 nm size. (c) Nanophox particle analysis of synthesized gold nanoparticles. (B) Ultraviolet-visible absorption spectra of gold nanoparticle-photosensitizer conjugate normalized to the maximum absorbance intensity. GNP has signature spectra at 540 nm due to surface plasmon resonance. MB shows absorption peaks at 662 nm and 613 nm. Conjugation of MB to GNP led to appearance of a small peak at 540 nm, intrinsic feature of GNP, which was absent in pure MB spectrum (a). TB spectrum enumerates peaks at 625 nm and 585 nm. GNP-TB conjugate is observed to exhibit an additional shoulder at 540 nm indicating conjugation of GNP to TB (b). Spectrum obtained for the mixture of GNP-MB and GNP-TB (c).
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pone.0131684.g001: Characterization of in-house synthesized GNPs and PS conjugation onto their surface.(A) Characterization of in-house synthesized gold nanoparticles. (a) Ultraviolet-visible- near infrared spectra of gold nanoparticles synthesized by exposing various amounts of Aloe vera extract to a fixed volume of HAuCl4 solution (10−3 M), keeping the final volume of reaction mixture 10 ml that was made up for each sample by increment in Aloe vera extract content for 24 h. Absorbance is seen to be increasing exhibiting two major peaks, a larger one at 540 nm, the characteristic peak of GNPs. (b) Representative TEM image of gold nanoparticles synthesized using Aloe vera leaf extract exhibiting a large population of spherical gold nanoparticles of approx. 10 nm size. (c) Nanophox particle analysis of synthesized gold nanoparticles. (B) Ultraviolet-visible absorption spectra of gold nanoparticle-photosensitizer conjugate normalized to the maximum absorbance intensity. GNP has signature spectra at 540 nm due to surface plasmon resonance. MB shows absorption peaks at 662 nm and 613 nm. Conjugation of MB to GNP led to appearance of a small peak at 540 nm, intrinsic feature of GNP, which was absent in pure MB spectrum (a). TB spectrum enumerates peaks at 625 nm and 585 nm. GNP-TB conjugate is observed to exhibit an additional shoulder at 540 nm indicating conjugation of GNP to TB (b). Spectrum obtained for the mixture of GNP-MB and GNP-TB (c).

Mentions: The gold nanoparticle synthesis was established by characteristic UV-visible spectra. As shown in Fig 1Aa, increase in the amount of plant extract results in an increase in the absorbance at 540 nm [24]. Further, microscopic analysis also ascertained GNPs synthesis. TEM analysis was performed with the nanogold solution harnessed after 24 h of incubation of HAuCl4 with 5 ml of Aloe vera leaf extract. Fig 1Ab shows representative TEM image displaying majorly spheroidal gold nanoparticles. Particles are seen to be in the size range of 10–20 nm. The size of the gold nanoparticle was also confirmed by nanophox analysis (Fig 1Ac).


Gold Nanoparticle-Photosensitizer Conjugate Based Photodynamic Inactivation of Biofilm Producing Cells: Potential for Treatment of C. albicans Infection in BALB/c Mice.

Sherwani MA, Tufail S, Khan AA, Owais M - PLoS ONE (2015)

Characterization of in-house synthesized GNPs and PS conjugation onto their surface.(A) Characterization of in-house synthesized gold nanoparticles. (a) Ultraviolet-visible- near infrared spectra of gold nanoparticles synthesized by exposing various amounts of Aloe vera extract to a fixed volume of HAuCl4 solution (10−3 M), keeping the final volume of reaction mixture 10 ml that was made up for each sample by increment in Aloe vera extract content for 24 h. Absorbance is seen to be increasing exhibiting two major peaks, a larger one at 540 nm, the characteristic peak of GNPs. (b) Representative TEM image of gold nanoparticles synthesized using Aloe vera leaf extract exhibiting a large population of spherical gold nanoparticles of approx. 10 nm size. (c) Nanophox particle analysis of synthesized gold nanoparticles. (B) Ultraviolet-visible absorption spectra of gold nanoparticle-photosensitizer conjugate normalized to the maximum absorbance intensity. GNP has signature spectra at 540 nm due to surface plasmon resonance. MB shows absorption peaks at 662 nm and 613 nm. Conjugation of MB to GNP led to appearance of a small peak at 540 nm, intrinsic feature of GNP, which was absent in pure MB spectrum (a). TB spectrum enumerates peaks at 625 nm and 585 nm. GNP-TB conjugate is observed to exhibit an additional shoulder at 540 nm indicating conjugation of GNP to TB (b). Spectrum obtained for the mixture of GNP-MB and GNP-TB (c).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4493101&req=5

pone.0131684.g001: Characterization of in-house synthesized GNPs and PS conjugation onto their surface.(A) Characterization of in-house synthesized gold nanoparticles. (a) Ultraviolet-visible- near infrared spectra of gold nanoparticles synthesized by exposing various amounts of Aloe vera extract to a fixed volume of HAuCl4 solution (10−3 M), keeping the final volume of reaction mixture 10 ml that was made up for each sample by increment in Aloe vera extract content for 24 h. Absorbance is seen to be increasing exhibiting two major peaks, a larger one at 540 nm, the characteristic peak of GNPs. (b) Representative TEM image of gold nanoparticles synthesized using Aloe vera leaf extract exhibiting a large population of spherical gold nanoparticles of approx. 10 nm size. (c) Nanophox particle analysis of synthesized gold nanoparticles. (B) Ultraviolet-visible absorption spectra of gold nanoparticle-photosensitizer conjugate normalized to the maximum absorbance intensity. GNP has signature spectra at 540 nm due to surface plasmon resonance. MB shows absorption peaks at 662 nm and 613 nm. Conjugation of MB to GNP led to appearance of a small peak at 540 nm, intrinsic feature of GNP, which was absent in pure MB spectrum (a). TB spectrum enumerates peaks at 625 nm and 585 nm. GNP-TB conjugate is observed to exhibit an additional shoulder at 540 nm indicating conjugation of GNP to TB (b). Spectrum obtained for the mixture of GNP-MB and GNP-TB (c).
Mentions: The gold nanoparticle synthesis was established by characteristic UV-visible spectra. As shown in Fig 1Aa, increase in the amount of plant extract results in an increase in the absorbance at 540 nm [24]. Further, microscopic analysis also ascertained GNPs synthesis. TEM analysis was performed with the nanogold solution harnessed after 24 h of incubation of HAuCl4 with 5 ml of Aloe vera leaf extract. Fig 1Ab shows representative TEM image displaying majorly spheroidal gold nanoparticles. Particles are seen to be in the size range of 10–20 nm. The size of the gold nanoparticle was also confirmed by nanophox analysis (Fig 1Ac).

Bottom Line: Photodynamic therapy (PDT) has been found to be effective in inhibiting biofilm producing organisms.We investigated the photodynamic effect of gold nanoparticle (GNP) conjugated photosensitizers against Candida albicans biofilm.The GNP-PS conjugate based PDT was found to effectively kill both C. albicans planktonic cells and biofilm populating hyphal forms.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, 202002, India.

ABSTRACT

Background: Photodynamic therapy (PDT) has been found to be effective in inhibiting biofilm producing organisms. We investigated the photodynamic effect of gold nanoparticle (GNP) conjugated photosensitizers against Candida albicans biofilm. We also examined the photodynamic efficacy of photosensitizer (PS) conjugated GNPs (GNP-PS) to treat skin and oral C. albicans infection in BALB/c mice.

Methods: The biomimetically synthesized GNPs were conjugated to photosensitizers viz. methylene blue (MB) or toluidine blue O (TB). The conjugation of PSs with GNPs was characterized by spectroscopic and microscopic techniques. The efficacy of gold nanoparticle conjugates against C. albicans biofilm was demonstrated by XTT assay and microscopic studies. The therapeutic efficacy of the combination of the GNP conjugates against cutaneous C. albicans infection was examined in mouse model by enumerating residual fungal burden and histopathological studies.

Results: The GNP-PS conjugate based PDT was found to effectively kill both C. albicans planktonic cells and biofilm populating hyphal forms. The mixture of GNPs conjugated to two different PSs significantly depleted the hyphal C. albicans burden against superficial skin and oral C. albicans infection in mice.

Conclusion: The GNP-PS conjugate combination exhibits synergism in photodynamic inactivation of C. albicans. The GNP conjugate based PDT can be employed effectively in treatment of cutaneous C. albicans infections in model animals. The antibiofilm potential of PDT therapy can also be exploited in depletion of C. albicans on medical appliances such as implants and catheters etc.

No MeSH data available.


Related in: MedlinePlus