Amornpun Sereemaspun

Effect of Gold Nanoparticle on Renal Cell: An Implication for Exposure Risk

Objective. The objective of this study was to evaluate the effect of gold nanoparticles on renal cell. Intervention. This study was performed as an experimental study. A mixture of gold nanoparticle solution and renal cell sediment was prepared and further analyzed. Results. This work revealed that after mixing the renal cell sediment with gold nanoparticle solution, gold nanoparticles can be seen within the renal cells. Conclusions. Conclusively, the gold nanoparticle can penetrate into renal cell. This is the first report on this observation, and further implies the possibility of other nanoparticle nephrotoxicity in the present nanomaterial era.

Expression of TNF-α, TGF-β, IP-10 and IL-10 mRNA in kidneys of hamsters infected with pathogenic Leptospira

Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira. Although several components of this organism have been identified, the molecular mechanisms underlying pathogenesis of this infectious disease are still poorly understood. Besides, direct injury by microbial factors, cytokines produced in response to infection have been proposed to be involved in pathogenesis of leptospirosis. In this study, cytokine gene expression in kidneys was investigated. Hamsters were injected with pathogenic Leptospira interrogans serovar Pyrogenes and were sacrificed on days 3, 5 and 7 after infection. RNA was extracted from kidney tissues. Real-time PCR was performed to demonstrate expression of TNF-alpha, TGF-beta, IP-10 and IL-10 mRNA in kidneys. TNF-alpha, TGF-beta and IP-10 expression could be demonstrated since day 3 post-infection whereas IL-10 expression was detected later on day 5. Leptospira infection resulted in not only expression of a proinflammatory cytokine, TNF-alpha, but also a T cell chemokine, IP-10. Detection of IP-10 suggested the involvement of T cell recruitment in the immune response or pathology in infected kidneys. Expressions of anti-inflammatory cytokines, TGF-beta and IL-10 were also observed. However, the level of TGF-beta expression was prominent since day 3 post-infection whereas IL-10 expression was clearly observed on day 5. Further experiments will provide additional information whether there is a correlation between the expression of these cytokines and pathologies found in an affected organ.

Detection of Leptospira in urine using anti-Leptospira-coated gold nanoparticles.

Serological assays for antibody detection have been widely used for Leptospirosis diagnosis. However, antibody is usually undetectable during the first week after infection. Detection of Leptospira DNA can be done by PCR but this technique requires special equipments and the cost is still relatively high. Here we demonstrate that gold nanoparticles can be used to facilitate Leptospira detection. Gold nanoparticles were coated with rabbit antibody specific to Leptospira interrogans serovar Bratislava and these coated particles were used to detect Leptospira in urine. Agglutination of gold particles indicated the presence of Leptospira in samples tested. The sensitivity of detection was 10 leptospires/ml. No agglutination was observed when anti-Leptospira-coated particles were tested with urine containing the organisms commonly found in urine such as Klebsiella pneumoniae, Escherichia coli and Enterococcus faecalis. This assay is very easy to perform and results could be observed with the naked eyes. Fresh or frozen urine samples could be used. The stability of antibody-coated particles was at least 2 months when kept at 4°C. In conclusion, we have demonstrated that the technique using antibody-coated gold nanoparticles is a promising tool for further validation as a rapid assay for Leptospirosis diagnosis.

Gold nanoparticle as an alternative tool for a urine pregnancy test.

OBJECTIVE The urine pregnancy test is an easily available diagnostic test in the present day and is routinely performed. The test is based on an immunochromatography technique. Here, we used an advanced nanomedicine technique for modification of the urine pregnancy test. MATERIALS AND METHODS The preparation of gold nanoparticle solution in this study followed the standard method. We performed an experiment on both pregnancy-positive and -negative urine samples. First, a mixture with an equal amount (500 microliters) of gold nanoparticle solution and urine sample was prepared. Then, it was further tested for pregnancy by the urine pregnancy test strip. RESULTS The pregnancy-positive mixture became pink, while the pregnancy-negative mixture became gray. The urine pregnancy test strip for a positive mixture had two lines, while the negative mixture had one line. CONCLUSION This application can help the diagnosis of pregnancy and can be an alternative method for a urine pregnancy test. To our knowledge, this is the first report on this application.

Effect of gold nanoparticle on the microscopic morphology of white blood cell

Background:  In medicine, there is limited knowledge on the toxicity of nanoparticles. In medicine, there has been limited knowledge on the effect of nanoparticles on the white blood cell.

Gold nanoparticle as an alternative tool for urine microalbumin test: the first world report.

Urine microalbuminuria is a common indicator for occult preclinical nephropathy. At present, urine microalbumin test is routinely performed by clinical chemistry test. Here, the authors used the advanced nanomedicine technique for modification of the urine microalbumin test. Materials and methods. The preparation of gold nanoparticle solution in this study was according to the standard method reported by Frenkel et al. The author performed an experiment on both microalbumin positive and negative urine samples. First, the mixture between equal amount (500 microliter) of gold nanoparticle solution and urine sample was prepared. Results. The microalbumin-positive mixture becomes purple, while the pregnancy-negative mixture becomes gray. Conclusion. This application can help the diagnosis of microalbumin and can be the alternative way for this urine microalbumin test. This is the first world report on this application.

Cytoprotective effect of glutaraldehyde erythropoietin on HEK293 kidney cells after silver nanoparticle exposure

The toxic effects from exposure to silver nanoparticles (AgNPs), which are broadly present in many consumer products, have long raised concerns. Many studies have focused on the mechanisms of nanosilver, which cause toxicity in human cells, but little is known about prevention of this type of injury. This study investigated the in vitro effects of glutaraldehyde erythropoietin (GEPO), a cytoprotective compound derived from erythropoietin, in terms of cell protection against AgNP-induced injury. HEK293 cells were pretreated with or without GEPO before administration of AgNPs. The protective effects of GEPO in this cell line were assessed by the percentage of viable cells, alterations of cell morphology, and the proliferative capability of the cells. In addition, we assessed the role of GEPO in lowering cellular oxidative stress and regulating expression of the anti-apoptotic protein Bcl2. The results showed rescue effects on the percentage of viable and proliferative cells among GEPO pretreated cells. Pretreatment with GEPO maintained the normal cell shape and ultrastructural morphology. Moreover, GEPO reduced the generation of reactive oxygen species in cells and activated expression of Bcl2, which are the major mechanisms in protection against cellular toxicity induced by AgNPs. In conclusion, our study showed that the cytotoxic effects from exposure to AgNPs can be prevented by GEPO.

Chitosan whisker grafted with oligo(lactic acid) nanoparticles via a green synthesis pathway: Potential as a transdermal drug delivery system

Chitosan whisker (CSWK) grafted with oligo(lactic acid) (OLA) nanoparticles (NPs) in water is developed to aid transferring therapeutic agents through the skin in a transdermal drug delivery system. Although several works in the past have shown grafting of poly(lactic acid) onto chitosan, the present work shows a green grafting system for the first time. The nano-sized CSWK provided effective conjugation of lactic acid even in a heterogeneous water-based system followed by polycondensation to form OLA. The OLA chain length is controlled by the lactic acid content and modulates the lipophilicity of CSWK-OLA. This fine tunes not only the size of the NPs but also the encapsulation efficiency of the hydrophobic drug lidocaine. A detailed chemical structure analysis, including the factors related to the development of NPs, is presented and extends the studies to the model drug encapsulation and delivery.

Chitosan-phenylalanine-mPEG nanoparticles: From a single step water-based conjugation to the potential allergen delivery system.

The chemical modification to obtain biocompatible chitosan (CS) nanoparticles for the application in biological system is still on expectation. By simply mixing CS with hydroxybenzotriazole (HOBt), the CS aqueous solution obtained allows a successful single step conjugation of both hydrophobic biomolecules, i.e. phenylalanine (Phe), and hydrophilic polymers, i.e. poly(ethylene glycol) methyl ether (mPEG), on CS, in water at room temperature. The CS-Phe-mPEG nanoparticles (20-50nm) exhibit positive charge leading to an entrapment of negatively charged house dust mite allergen (HDM) extract (Dermatophagoides pteronyssinus). The HDM-entrapped CS-Phe-mPEG shows biocompatibility as evidenced from the cell viability, the ROS (reactive oxygen species) reduction, and the HaCaTs proliferation. The clinical implementation on the healthy- and HDM-allergic volunteers indicates that the HDM-entrapped CS-Phe-mPEG stimulates cell-mediated immune response in peripheral blood mononuclear cells (PBMCs) and favors T cell immune response as seen from the reduction of interferon-(IFN)-γ and interleukin-(IL)-10 in the PBMCs of the HDM-allergic volunteers.

Immunogold-agglutination assay for direct detection of HPV-16 E6 and L1 proteins from clinical specimens

HPV-16 infection is the most common cause of cervical cancer. As HPV-16 transforms the cell, E6 oncoprotein is over-expressed. Therefore, molecular detection of HPV-16 E6 mRNA is now being used for diagnosis and prediction of cancer development. Besides detecting E6 mRNA, a rapid lateral flow detecting the E6 protein using enzyme immunoassay is also now on market with a sensitivity of 53.5% for cervical intraepithelial neoplasia (CIN)-3 or more severe (CIN-3+). Here, an immunogold-agglutination assay was developed to detect not only HPV-16 E6 protein but also L1, a major capsid protein found in the productive stage of the virus. Evaluation of this test using HPV-16 DNA positive cervical samples showed that the HPV-16 E6 immunogold-agglutination assay results correlated well with the progression of the cervical lesions, i.e., 10.34% of CIN-1, 68.75% of CIN-3 and 80% of cancer (CaCx) and none for healthy normal samples. Interestingly, the HPV-16 L1 protein was found in most of the cases with cancer indicating the possibility of virion production. Immunogold-agglutination assay for E6 protein is simpler, easier to be performed with a sensitivity of 73.1% for CIN-3+ suggesting a good method for laboratory diagnostic use.