Bridging the Gap with Nanoparticles: A Novel Approach.

Two-piece implants unavoidably present a microgap between the implant and the abutment interface. Although numerous modalities have been attempted to overcome this situation, the implant abutment interface still remains a critical point for microbial colonization, which starts an inflammatory cascade of events eventually compromising the implants. Throughout our life, cells in all biological systems are unprotected to oxidative stress leading to the formation of Reactive oxygen species which is of concern when it comes to placing implants in patients who are periodontally compromised. This necessitates the development of alternative therapeutic modalities, which could counteract as well as prevent the microbial overload and ROS generation thereby improving the longevity of implants. To evaluate and assess the antibacterial, antioxidant and anti inflammatory effectiveness of quercetin-loaded titanium nanocomposites as coatings over healing abutments. Quercetin-loaded titanium nanocomposites were synthesized using green synthesis and confirmation was done using UV spectroscopy. Healing abutments were coated with the formulated nanocomposites, an intra-oral environment was simulated by thermocycling. Their antibacterial, antioxidant, anti-inflammatory, and cytotoxicity were assessed using standard tests. Healing abutments were coated with the formulated nanocomposites, an intra-oral environment was simulated by thermocycling. They showed potent antibacterial, antioxidant, and anti-inflammatory properties, which could prove beneficial in a variety of clinical scenarios in which there is a high risk for implant failure during early osseointegration.

Green synthesis of calcium hydroxide-coated silver nanoparticles using Andrographis paniculata and Ocimum sanctum Linn. leaf extracts: An antimicrobial and cytotoxic activity.

Context: Silver is known for its antibacterial properties since ages. As nanoparticles have smaller size and greater surface area, silver has been utilized in the form of nanoparticles to enhance its antibacterial properties. Calcium hydroxide is a well-known intracanal medicament and serves as a gold standard for root canal disinfection. Using herbal extracts as reducing agents for nanoparticle synthesis appears to be an ecofriendly approach. Aim: The aim of this study was to synthesize calcium hydroxide-based silver nanoparticles using herbs as reducing agents and to test the cytotoxic levels and antimicrobial activity against oral microbes. Materials and Methods: The calcium hydroxide-based silver nanoparticles were synthesized using the leaves of Andrographis paniculata and Ocimum sanctum Linn. Various properties of the synthesized nanoparticles were also characterized by ultraviolet (UV) spectrophotometer analysis, transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier-transform infrared spectroscopy (FTIR) analysis. The cytotoxic effects of these nanoparticles were analyzed using brine shrimp and MTT assay. Antimicrobial activity was assessed by measuring the zone of inhibition. The statistical analysis was done using parametric independent t-test. P value was set at < 0.05. Results: The calcium hydroxide-based silver nanoparticles were successfully synthesized and were confirmed by UV spectrophotometer analysis, TEM, X-ray diffraction (XRD), and FTIR analysis and showed a minimal cytotoxic effect. They also showed a good antimicrobial activity and a remarkable antifungal activity. Conclusions: The green synthesis of CaOHAgNPs yielded an effective nanoparticle preparation that could be used against common oral pathogens as a potential therapeutic agent in the form of root canal irrigant or intracanal medicament in the field of dentistry.

Oligonucleotide therapy: An emerging focus area for drug delivery in chronic inflammatory respiratory diseases.

Oligonucleotide-based therapies are advanced novel interventions used in the management of various respiratory diseases such as asthma and Chronic Obstructive Pulmonary Disease (COPD). These agents primarily act by gene silencing or RNA interference. Better methodologies and techniques are the need of the hour that can deliver these agents to tissues and cells in a target specific manner by which their maximum potential can be reached in the management of chronic inflammatory diseases. Nanoparticles play an important role in the target-specific delivery of drugs. In addition, oligonucleotides also are extensively used for gene transfer in the form of polymeric, liposomal and inorganic carrier materials. Therefore, the current review focuses on various novel dosage forms like nanoparticles, liposomes that can be used efficiently for the delivery of various oligonucleotides such as siRNA and miRNA. We also discuss the future perspectives and targets for oligonucleotides in the management of respiratory diseases.

Quercetin conjugated superparamagnetic magnetite nanoparticles for in-vitro analysis of breast cancer cell lines for chemotherapy applications.

The magnetic nanoparticles attract increasing interest due to their opportunities in cancer therapy and used as drug carriers for several other diseases. The present study investigates the quercetin conjugated superparamagnetic Fe3O4 nanoparticles for in-vitro analysis of breast cancer cell lines for chemotherapy. A simple precipitation method was used to prepare the dextran coated Fe3O4 nanoparticles and the anticancer flavonoid quercetin was conjugated on the surface via carboxylic/amine group using nanoprecipitation method. The structural, morphological and the magnetic properties of the prepared materials were studied by using X-ray diffractometer (XRD), Fourier transformed infer-red spectrometer (FTIR), transmission electron microscope (TEM) and vibrating sample magnetometer (VSM). The MTT (3-(4,5-dimethylthiahiazol-2-yl)-2,5-diphenyl tetrazolium) assay of dextran coated Fe3O4 nanoparticles did not exhibit notable toxicity against MCF7 cells, whereas the cytotoxicity of quercetin conjugated Fe3O4 nanoparticles increased significantly in comparison with pure quercetin. The incubation of MCF-7 cells with quercetin conjugated Fe3O4 nanoparticles (QCMNPs) shows significant changes in cellular morphology observed through fluorescent microscopy. The results validate the prepared quercetin conjugated Fe3O4 nanoparticles are promising anticancer agents for targeted drug delivery.

Immunization with baculovirus displayed H6 hemagglutinin vaccine protects mice against lethal H6 influenza virus challenge.

Low pathogenic influenza viruses of H6 hemagglutinin (HA) subtype have a high prevalence among aquatic and domestic birds and have caused outbreaks in poultry worldwide. The first human infection with wild avian influenza H6N1 virus was reported in Taiwan and these subtype viruses may continue to evolve and accumulate changes which increasing the potential risk of human-to-human transmission. To develop a vaccine against influenza viruses of the H6 subtype, we displayed the HA gene on the baculovirus surface (Bac-HA), and studied its vaccine efficacy against a lethal challenge with mouse-adapted RG-H6(Shorebird) virus carrying the H6 HA gene from A/shorebird/DE/12/2004 (H6N8) virus and 7 genes from A/Puerto Rico/8/1934 (H1N1) virus. Immunization with 256 HA units of Bac-HA via the intranasal route triggered HA-specific serum and mucosal antibodies in mice besides increased HA inhibition titers compared to mice immunized subcutaneously. Moreover, we observed an increase in cellular immune response (IL-4) and improved in vitro neutralization activity in the mice immunized intranasally with live Bac-HA compared to mice immunized with inactivated influenza virus (IV). Interestingly, Bac-HA intranasal immunized mice showed one fold higher neutralization titer against heterologous H6 influenza virus compared to inactivated IV immunized mice. In addition, the live Bac-HA, administered through either immunization route, as well as the adjuvanted inactivated Bac-HA, administered subcutaneously, conferred 100% protection to mice challenged with homologous mouse-adapted RG-H6(Shorebird) virus. The reduction in viral titers and extend of histopathological changes of Bac-HA immunized mice lungs further demonstrated the protective efficacy of Bac-HA. Hence, the recombinant baculovirus subunit vaccine is an alternative candidate against H6 subtypes that could be propagated and administered with minimal biosafety concerns.

Rhinolithiasis due to supernumerary ectopic tooth: very rare case.

An ectopic supernumerary tooth causing the formation of rhinolith was never reported before in the medical literature. A 30 years old male patient presented to our hospital with one sided nasal obstruction, recurrent epistaxis, and nasal discharge. Anterior rhinoscopy, nasal endoscopy and X-ray paranasal sinuses revealed a rhinolith in the left nasal cavity. Preoperative evaluation and post operative examination of the specimen proved that the nidus of rhinolith was a supernumerary ectopic tooth.

Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton).

Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita, and the brain of catla, Catla catla, respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 degrees C with an optimum of 28 degrees C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 degrees C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp. were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.

Natural aquatic insect carriers of Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV).

Five different species of aquatic insects were collected from nursery ponds containing the freshwater prawn Macrobrachium rosenbergii infected with Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV). The insects were screened as potential natural carriers of MrNV and XSV. RT-PCR (reverse transcription polymerase chain reaction) analysis gave positive results for MrNV and XSV in Belostoma sp., Aesohna sp., Cybister sp. and Notonecta sp., and negative results for Nepa sp. An Aedes albopictus mosquito cell line (C6/36) was used for infectivity assays, with viral inoculum prepared from the aquatic insects, since C6/36 cells have recently been shown to be susceptible to infection with MrNV and XSV. The C6/36 cells were harvested 4 d post-challenge for examination by electron microscopy. This revealed aggregation of viral particles throughout the cytoplasm for cells challenged with inocula from all the insect species except Nepa sp. Our results indicate that several aquatic insect species may present a risk for MrNV and XSV transmission to M. rosenbergii.

Silencing VP28 gene of white spot syndrome virus of shrimp by bacterially expressed dsRNA.

An in vivo expression system to produce large amounts of virus-derived dsRNAs in bacteria to provide a practical control of white spot syndrome virus (WSSV) in shrimp was developed. The bacterially synthesized dsRNA specific to VP28 gene of WSSV promoted gene-specific interference with the WSSV infection in shrimp. Virus infectivity was significantly reduced in WSSV-challenged shrimp injected with VP28-dsRNA and 100% survival was recorded. The inhibition of the expression of WSSV VP28 gene in experimentally challenged animals by VP28-dsRNA was confirmed by RT-PCR and Western blot analyses. Furthermore, we have demonstrated the efficacy of bacterially expressed VP28-dsRNA to silence VP28 gene expression in SISK cell line transfected with eukaryotic expression vector (pcDNA3.1) inserted with VP28 gene of WSSV. The expression level of VP28 gene in SISK cells was determined by fluorescent microscopy and ELISA. The results showed that the expression was significantly reduced in cells transfected with VP28dsRNA, whereas the cells transected with pcDNA-VP28 alone showed higher expression. The in vivo production of dsRNA using prokaryotic expression system could be an alternative to in vitro method for large-scale production of dsRNA corresponding to VP28 gene of WSSV for practical application to control the WSSV in shrimp farming.

Protective efficiency of DNA vaccination in Asian seabass (Lates calcarifer) against Vibrio anguillarum.

Vibriosis is one of the most prevalent fish diseases caused by bacteria belonging to the genus Vibrio. Vibriosis caused by Vibrio anguillarum produces a 38-kDa major outer membrane porin protein (OMP) for biofilm formation and bile resistant activity. The gene encoding the porin was used to construct DNA vaccine. The protective efficiency of such vaccine against V. anguillarum causing acute vibrio haemorrhagic septicaemia was evaluated in Asian seabass (Lates calcarifer Bloch), a common species of the Indian coast and a potential resource for the aquaculture industry. In vitro protein expression of porin gene was determined by fluorescent microscopy after transfection of seabass kidney cell line (SISK). Fish immunized with a single intramuscular injection of 20 microg of the OMP38 DNA vaccine showed significant serum antibody levels in 5th and 7th weeks after vaccination, compared to fish vaccinated with the control eukaryotic expression vector pcDNA3.1. Asian seabass vaccinated with the OMP38 DNA vaccine was challenged with pathogenic V. anguillarum by intramuscular injection. A relative percent survival (RPS) rate of 55.6% was recorded. Bacterial agglutination and serum complement activity was analysed by using DNA vaccinated seabass serum above 80% of analysed strain was killed at the highest agglutination titre. Histopathological signs of V. anguillarum challenged fish were observed in around 45% of pVAOMP38, 90% of PBS and 87% of pcDNA3.1-vaccinated control fish. The results indicate that L. calcarifer vaccinated with a single dose of DNA plasmid encoding the major outer membrane protein shows moderate protection against acute haemorrhagic septicaemia and mortality by V. anguillarum experimental infection.