Cisplatin-loaded albumin nanoparticle and study their internalization effect by using ß-cyclodextrin.

The present study with aim at enhancing the therapeutic and anti-cancer properties of cisplatin (CPT)-loaded bovine serum albumin (BSA) nanoparticles. The BSA nanoparticles containing CPT (CPT-BSANPs) were successfully prepared by the desolvation technique. The physicochemical characterization of the CPT-BSANPs were used by Fourier transformed infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The particle size of CPT-BSANPs was found less than 200 nm with 75.02 ± 0.15% entrapment efficiency (EE), while zeta potential and PDI were -17.6 mV and 0.2, respectively. In vitro release behavior of the CPT from the carrier suggests that about 64% of the drug gets released after 48 hrs. The anti-cancer activities of the CPT-BSANPs were tested on MCF-7 cell lines. Our studies show that CPT-BSANPs nanoparticles showed specific targeting and enhanced cytotoxicity to MCF-7 cells when compared to the bare CPT. Thus results suggest that CPT-BSANPs fallowed caveolae-mediated endocytosis, it may become better option for intracellular delivery of anticancer drug.

Studies on antimicrobial and wound healing applications of gauze coated with CHX-Ag hybrid NPs.

In this study, chlorhexidine (CHX)-silver (Ag) hybrid nanoparticles (NPs) coated gauze was developed, and their bactericidal effect and in vivo wound healing capacities were tested. A new method was developed to synthesise the NPs, wherein Ag nitrate mixed with sodium (Na) metaphosphate and reduced using Na borohydride. Finally, CHX digluconate was added to form the hybrid NPs. To study the antibacterial efficacy of particles, the minimal inhibition concentration and biofilm degradation capacity against Gram-positive and Gram-negative bacteria was studied using Escherichia coli and Staphylococcus aureus. The results indicated that the NP inhibited biofilm formation and was bactericidal as well. The gauze was doped with NPs, and its wound healing property was evaluated using mice model. Results indicated that the wound healing process was fastened by using the NPs gauze doped with NPs without the administration of antibiotics.

Ketoconazole-conjugated ZnO nanoparticles based semi-solid formulation and study their impacts on skin disease.

In this study, the ketoconazole-conjugated zinc oxide (ZnO) nanoparticles were prepared in a single-step approach using dextrose as an intermediate compound. The physical parameters confirmed the drug conjugation with ZnO and their size was around 70-75 nm. The drug loading and in vivo drug release studies indicated that the -CHO group from the dextrose increase the drug loading up to 65% and their release kinetics were also studied. The anti-fungal studies indicated that the prepared nanoparticles exhibit strong anti-fungal activity and the minimum concentration needed is 10 mg/ml. The nanoparticles loaded semi-solid gel was prepared using carbopol, methylparaben, propyl paraben and propylene glycol. The in vitro penetration of the ketoconazole-conjugated nanoparticles was studied using the skin. The results indicated that the semi-solid gel preparations influenced the penetration and also favoured the accumulation into the skin membrane. The veterinary clinical studies indicated that the prepared gel is highly suitable for treatment of Malassezia.

Chlorhexidine-calcium phosphate nanoparticles - Polymer mixer based wound healing cream and their applications.

In this work, we developed a wound healing cream composed of two different polymers, namely chitosan and gelatin with chlorhexidine along with calcium phosphate nanoparticles. The physicochemical properties of the prepared cream were investigated based on SEM, EDX, Raman, FTIR and the results indicated that the cream contained gelatin, chitosan, calcium phosphate nanoparticles and chlorhexidine. The maximum swelling ratio studies indicated that the ratio was around of 52±2.2 at pH7.4 and the value was increased in acidic and alkaline pH. The antimicrobial activity was tested against bacteria and the results indicated that, both chlorhexidine and the hybrid cream devoid of chlorhexidine exhibited antimicrobial activity but the chlorhexidine impregnated cream showed three fold higher antimicrobial activity than without chlorhexidine. In vivo wound healing promoting activities of hybrid cream containing 0.4mg/L chlorhexidine were evaluated on surgically induced dermal wounds in mice. The results indicated that the cream with incorporated chlorhexidine significantly enhanced healing compared with the control samples. For the field validations, the veterinary clinical animals were treated with the cream and showed enhanced healing capacity. In conclusion, a simple and efficient method for design of a novel wound healing cream has been developed for veterinary applications.

In ovo delivery of Newcastle disease virus conjugated hybrid calcium phosphate nanoparticle and to study the cytokine profile induction.

In this report, the hybrid calcium phosphate (CaP) nanoparticles were synthesized and functionalized with Newcastle disease virus (NDV). These nanoparticles were synthesized by a combination of co-precipitation and polymerization process and functionalized with amino propyl triethoxy silane before coupling to NDV. The 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT) assay of chicken spleen cells incubated with these nanoparticles indicated that, these particles did not exert any significant cytotoxicity. The effects of hybrid CaP nanoparticles on cell cycle were assayed using a flow cytometer. The results demonstrated that the cell viability and proliferation capacity of spleen cells were not affected by hybrid CaP nanoparticles compared with their control cells. The hybrid CaP nanoparticles were characterized by scanning/transmission electron microscopy (SEM/TEM); Fourier transformed infrared spectroscopy (FTIR), X-ray diffraction patterns (XRD), Raman spectroscopy and energy-dispersive X-ray spectroscopy (EDX). These methods revealed that NDV was successfully conjugated on nanoparticles. The ability of the hybrid CaP nanoparticles to induce different cytokine mRNAs in the spleen cells of 18-day old embryonated chicken eggs (ECEs) was studied by quantitative real time polymerase chain reaction (qRT-PCR). NDV conjugated particles induced a high expression of Th1 cytokines such as interferon (IFN)-α, tumor necrosis factor (TNF)-α of and Th2 cytokines, interleukin (IL) 6 and IL-10. Uncoupled NDV induced only Th1 cytokines, IFN-α, INF-γ and TNF-α. The hybrid particles alone did not induce any cytokines. This confirmed that nanoparticle coupling could induce differential cytokine profiles and hence can be used as an alternate strategy to direct favorable immune responses in animals or chickens using appropriate vaccination carrier.

Rapid determination of hydrogen peroxide produced by Lactobacillus using enzyme coupled rhodamine isocyanide/calcium phosphate nanoparticles.

A sensitive method for detecting hydrogen peroxide (H2O2) using rhodamine isocyanide incorporated calcium phosphate nanoparticles (Rho/CaP) was developed. The synthesized nanoparticles were characterized based on transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transformed infrared spectroscopy (FTIR), Raman spectroscopy, energy-dispersive X-ray spectroscopy (EDX) and X-ray diffraction patterns (XRD). To study the application, the nanoparticles were functionalized with horse radish peroxidase (HRP) based on aminopropyl triethoxy silane (APTES) and used as tools to detect H2O2. The detection strategy was based on fluorescence quenching or colorimetric detection. The enzyme immobilized nanoparticles were titrated with different concentrations of H2O2 and a fixed concentration of O-phenylenediamine (OPD). The HRP conjugated Rho/CaP strongly catalyzed H2O2 oxidation of OPD that caused fluorescence quenching at 575 nm. For colorimetric detection, the OPD product was read at 492 nm. In the fluorescence quenching assay, the minimum detectable concentration was ~1 pmol in contrast to ~5 nmol in the colorimetric assay. The minimum detectable concentration by visual detection was ~500 nmol. The specificity of the developed assay method was examined with different interferences which did not produce any significant response. This assay was applied, along with a commercially available kit to compare the H2O2 production capacities of different Lactobacillus strains. The results indicated that the developed assay and commercially available kit methods were highly correlated. The fluorescence quenching kinetics is also discussed.

Formulation of Newcastle disease virus coupled calcium phosphate nanoparticles: an effective strategy for oculonasal delivery to chicken.

In this report, calcium phosphate (CaP) nanoparticles were synthesized by continuous flow method using ß-cyclodextrin (ß-CD) as a medium and functionalized with amino propyl triethoxy silane (APTES). The blood biocompatibility of the nanoparticles was assessed using the whole blood haemolysis, erythrocytes haemolysis and erythrocyte aggregation tests. Based on the results, we found that the synthesized ß-CD-CaP nanoparticles did not cause any remarkable toxic effect. The 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT) assay of chicken peripheral blood mononucleated cells (PBMCs) incubated with these nanoparticles indicated that these particles did not exert any significant cytotoxicity. The aminosilane functional group modified ß-CD-CaP was used as tool for coupling of Newcastle disease virus (NDV). The NDV conjugated nanoparticles were confirmed by using Fourier transformed infrared spectroscopy, X-ray diffraction patterns, Raman spectroscopy differential scanning calorimetry and energy-dispersive X-ray spectroscopy. Immunogenicity trials in chickens proved that ß-CD-CaP-NDV used as a vaccine was better than the commercial vaccine when given oculonasally during the first 2 weeks post vaccination. The birds vaccinated with the above nano-NDV vaccine were completely protected against virulent NDV challenge. This study confirms that the oculonasal ß-CD-CaP-NDV delivery of vaccines is a potential method for enhancing the immune responses of existing commercial vaccines.

Development of antibiotic selection kit towards veterinary applications using glycine passivated magnetic particles.

Glycine functionalized (Gly/Fe3O4) and non-functionalized (Fe3O4) magnetic particles were synthesized in an autoclave and characterized by transmission electron microscopy (TEM), Fourier transformed infrared spectroscopy (FTIR), vibrating sample magnetometer (VSM), energy-dispersive X-ray spectroscopy (EDX), differential scanning calorimetry (DSC), X-ray diffraction patterns (XRD) and zeta potential. The size of the both these particles were in the range of 220-230 nm but the shape of the Gly/Fe3O4 particles was hexagonal in contrast to the spherical shape of Fe3O4 particles. The particle characterization tests confirmed that glycine was functionalized on the Gly/Fe3O4 particles, they were positively charged and possessed strong magnetic property. These particles possessed the ability to bind to bacteria such as Escherichia coli, Streptococcus and Staphylococcus in the range of 72-90%. They were used to entrap bacteria from clinical mastitic milk samples from cows. The entrapped bacteria of the above species from these samples were isolated and used individually in the conventional disc-diffusion method of antibiotic susceptibility determination. The results were compared with that of the bacterial species isolated directly from the mastitic milk samples and were found to be 100% concordant (n=25). The developed portable antibiotic selection kit was tested with twenty five samples of mastitic milk. The results indicated that, antibiotic resistant bacteria turned the methylene blue in to white color while the bacteria that were killed (sensitive) retained the blue color of the dye. Thus the right choice of the antibiotic to treat cows with mastitis could be determined based on the naked eye. In conclusion, the kit gave quicker results, was easy to assay and read and can be 'farm-gate' applicable than the presently available conventional method.