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Facile synthesis of biocompatible gold nanoparticles from Vites vinefera and its cellular internalization against HBL-100 cells.

Amarnath K, Mathew NL, Nellore J, Siddarth CR, Kumar J - Cancer Nanotechnol (2011)

Bottom Line: Current discovery demonstrates the rapid formation of gold nanoparticles with the phytochemicals present in grapes, which serve a dual role as synergistic reducing agents to reduce gold salts into gold nanoparticles and also as stabilizers to provide a robust coating on the gold nanoparticles in a single step.In addition, the grape-generated gold nanoparticles (GAuNPs, GSH-GAuNPs, LA-GAuNPs) have demonstrated remarkable affinity towards human breast cancer cells (HBL-100) in the present study.Other than gold salts, no "manmade" chemicals are used in this truly biogenic, green nanotechnological process which thereby paves the way for outstanding opening for their application in molecular imaging and cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biochemistry, Dental College & Hospitals, Sathyabama University, Chennai, 600119 Tamil Nadu India.

ABSTRACT

The remarkable health benefits of the chemical cocktails occluded within Vites vinefera (grapes) have been broadly used as dietary supplements and as natural pharmaceuticals in the treatment of various diseases including human cancer. Current discovery demonstrates the rapid formation of gold nanoparticles with the phytochemicals present in grapes, which serve a dual role as synergistic reducing agents to reduce gold salts into gold nanoparticles and also as stabilizers to provide a robust coating on the gold nanoparticles in a single step. Furthermore, the grape-generated gold nanoparticles (GAuNPs), have demonstrated remarkable in vitro stability on specific functionalization with peptides (GSH) and thiol-containing compounds (lipoic acid) followed by the induction of cell-specific response. In addition, the grape-generated gold nanoparticles (GAuNPs, GSH-GAuNPs, LA-GAuNPs) have demonstrated remarkable affinity towards human breast cancer cells (HBL-100) in the present study. These studies thus signified the cellular internalization of GAuNPs and its conjugates by transmission electron microscopy through endocytosis into cancer cells. Notably, at higher concentration of gold nanoparticles conjugate, there was an asymmetric accumulation of gold nanoparticles in the periphery of the cell nucleus of the HBL-100 cells which was confirmed by fluorescence microscopy. Other than gold salts, no "manmade" chemicals are used in this truly biogenic, green nanotechnological process which thereby paves the way for outstanding opening for their application in molecular imaging and cancer therapy.

No MeSH data available.


Related in: MedlinePlus

Phase contrast microscopic pictures of HBL-100 cells a untreated and treated b with 500 μM GAuNPs, c GSH-GAuNPs, and d liopic acid-GAuNPs
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Fig6: Phase contrast microscopic pictures of HBL-100 cells a untreated and treated b with 500 μM GAuNPs, c GSH-GAuNPs, and d liopic acid-GAuNPs

Mentions: Characterization of in vitro nanoparticle uptake and localization is intrinsically linked to cytotoxicological studies because uptake provides evidence of nanoparticle–cell interaction, wherein the delicate intracellular machinery is exposed to nanoparticles. Compared to in vivo studies, in vitro studies benefit from being faster, lower cost, allowing greater control, and minimizing ethical concerns by reducing the number of laboratory animals required for testing. The most commonly used in vitro assessment techniques generally evaluate either viability (live/dead ratio) or toxicity mechanism. Herein, the major viability-based assays are organized into the categories of proliferation, necrosis, or apoptosis DNA damage detection techniques. In our present communication, the cytotoxicty of GAuNPs, GSH-GAuNPs, and LA-GAuNPs under in vitro conditions in HBL-100 cells was examined using two cytotoxicity markers, including MTT reduction, and LDH leakage is used to study the effects of gold nanopatricle conjugates on cellular viability. To check the cytotoxicity of cells treated with GAuNPs (10–150 μL), GSH-GAuNPs (10–150 μL), and LA-GAuNPs (10–150 μL) for 24 h, and a number of viable cells were enumerated by colorimetric MTT assay. After 24 h of post-treatment, HBL-100 cells showed excellent viability even up to 150 μL of GAuNPs, GSH-GAuNPs, and LA-GAuNPs (Fig. 5). Results of MTT assays clearly revealed the cytotoxic effect of GAuNPs in a dose-dependent manner for HBL-100 cell lines and LA-GAuNPs exerted slightly better cytotoxic effect towards HBL-100 cells in comparison to GSH-GAuNPs. Although GAuNPs also showed cytotoxicity towards cancer cells, the effect was much less compared to GSH-GAuNPs and LA-GAuNPs. Treatment with 150 μL or higher doses of GAuNPs and GSH-GAuNPs and LA-GAuNPs to HBL-100 cell increased LDH leakage into culture media (Fig. 6), signifying an AUNP-induced compromise of plasma membrane integrity, and the IC50 of AgNPs was 150 μL. Henceforth, the release of LDH (Fig. 6) in our study is in agreement with the excellent viability of the cells treated with GAuNPs and GSH-GAuNPs and LA-GAuNPs as proven by MTT assay. It is also important to recognize that a vast majority of gold (I) and gold (III) compounds exhibit varying degrees of cytotoxicity to a variety of cells (Basset et al. 2003; Hamer 2007). GAuNPs pretreatment at a concentration of 150 μL reduced the LDH leakage to a minimum, and this concentration is used in subsequent studies.Fig. 6


Facile synthesis of biocompatible gold nanoparticles from Vites vinefera and its cellular internalization against HBL-100 cells.

Amarnath K, Mathew NL, Nellore J, Siddarth CR, Kumar J - Cancer Nanotechnol (2011)

Phase contrast microscopic pictures of HBL-100 cells a untreated and treated b with 500 μM GAuNPs, c GSH-GAuNPs, and d liopic acid-GAuNPs
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4544069&req=5

Fig6: Phase contrast microscopic pictures of HBL-100 cells a untreated and treated b with 500 μM GAuNPs, c GSH-GAuNPs, and d liopic acid-GAuNPs
Mentions: Characterization of in vitro nanoparticle uptake and localization is intrinsically linked to cytotoxicological studies because uptake provides evidence of nanoparticle–cell interaction, wherein the delicate intracellular machinery is exposed to nanoparticles. Compared to in vivo studies, in vitro studies benefit from being faster, lower cost, allowing greater control, and minimizing ethical concerns by reducing the number of laboratory animals required for testing. The most commonly used in vitro assessment techniques generally evaluate either viability (live/dead ratio) or toxicity mechanism. Herein, the major viability-based assays are organized into the categories of proliferation, necrosis, or apoptosis DNA damage detection techniques. In our present communication, the cytotoxicty of GAuNPs, GSH-GAuNPs, and LA-GAuNPs under in vitro conditions in HBL-100 cells was examined using two cytotoxicity markers, including MTT reduction, and LDH leakage is used to study the effects of gold nanopatricle conjugates on cellular viability. To check the cytotoxicity of cells treated with GAuNPs (10–150 μL), GSH-GAuNPs (10–150 μL), and LA-GAuNPs (10–150 μL) for 24 h, and a number of viable cells were enumerated by colorimetric MTT assay. After 24 h of post-treatment, HBL-100 cells showed excellent viability even up to 150 μL of GAuNPs, GSH-GAuNPs, and LA-GAuNPs (Fig. 5). Results of MTT assays clearly revealed the cytotoxic effect of GAuNPs in a dose-dependent manner for HBL-100 cell lines and LA-GAuNPs exerted slightly better cytotoxic effect towards HBL-100 cells in comparison to GSH-GAuNPs. Although GAuNPs also showed cytotoxicity towards cancer cells, the effect was much less compared to GSH-GAuNPs and LA-GAuNPs. Treatment with 150 μL or higher doses of GAuNPs and GSH-GAuNPs and LA-GAuNPs to HBL-100 cell increased LDH leakage into culture media (Fig. 6), signifying an AUNP-induced compromise of plasma membrane integrity, and the IC50 of AgNPs was 150 μL. Henceforth, the release of LDH (Fig. 6) in our study is in agreement with the excellent viability of the cells treated with GAuNPs and GSH-GAuNPs and LA-GAuNPs as proven by MTT assay. It is also important to recognize that a vast majority of gold (I) and gold (III) compounds exhibit varying degrees of cytotoxicity to a variety of cells (Basset et al. 2003; Hamer 2007). GAuNPs pretreatment at a concentration of 150 μL reduced the LDH leakage to a minimum, and this concentration is used in subsequent studies.Fig. 6

Bottom Line: Current discovery demonstrates the rapid formation of gold nanoparticles with the phytochemicals present in grapes, which serve a dual role as synergistic reducing agents to reduce gold salts into gold nanoparticles and also as stabilizers to provide a robust coating on the gold nanoparticles in a single step.In addition, the grape-generated gold nanoparticles (GAuNPs, GSH-GAuNPs, LA-GAuNPs) have demonstrated remarkable affinity towards human breast cancer cells (HBL-100) in the present study.Other than gold salts, no "manmade" chemicals are used in this truly biogenic, green nanotechnological process which thereby paves the way for outstanding opening for their application in molecular imaging and cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biochemistry, Dental College & Hospitals, Sathyabama University, Chennai, 600119 Tamil Nadu India.

ABSTRACT

The remarkable health benefits of the chemical cocktails occluded within Vites vinefera (grapes) have been broadly used as dietary supplements and as natural pharmaceuticals in the treatment of various diseases including human cancer. Current discovery demonstrates the rapid formation of gold nanoparticles with the phytochemicals present in grapes, which serve a dual role as synergistic reducing agents to reduce gold salts into gold nanoparticles and also as stabilizers to provide a robust coating on the gold nanoparticles in a single step. Furthermore, the grape-generated gold nanoparticles (GAuNPs), have demonstrated remarkable in vitro stability on specific functionalization with peptides (GSH) and thiol-containing compounds (lipoic acid) followed by the induction of cell-specific response. In addition, the grape-generated gold nanoparticles (GAuNPs, GSH-GAuNPs, LA-GAuNPs) have demonstrated remarkable affinity towards human breast cancer cells (HBL-100) in the present study. These studies thus signified the cellular internalization of GAuNPs and its conjugates by transmission electron microscopy through endocytosis into cancer cells. Notably, at higher concentration of gold nanoparticles conjugate, there was an asymmetric accumulation of gold nanoparticles in the periphery of the cell nucleus of the HBL-100 cells which was confirmed by fluorescence microscopy. Other than gold salts, no "manmade" chemicals are used in this truly biogenic, green nanotechnological process which thereby paves the way for outstanding opening for their application in molecular imaging and cancer therapy.

No MeSH data available.


Related in: MedlinePlus