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1.
Proc Inst Mech Eng H ; 227(5): 510-22, 2013 May.
Article in English | MEDLINE | ID: mdl-23637261

ABSTRACT

The goal of the experiment outlined in this article is to improve upon noncemented methods of arthroplasty for clinical application in elderly patients. This was done by determining whether titanium implants with a novel nanostructured zirconia surface, which was created by ion beam-assisted deposition, would prevent impaired osseointegration of intramedullary implants in 1-year-old rats receiving a protein-deficient diet. Specifically, we asked whether the implant with the nanostructured zirconia surface would increase expression of markers of bone maturation within the remodeling of peri-implant woven bone. The control implants, which were made of commercially pure titanium, had a polished surface ex vivo but are known to acquire a microstructured titania surface in vivo. Ten 1-year-old rats received experimental implant (group A) and 10 had control (group B) implants. Ten 3-month-old rats received normal protein diet and the control implant (group C). Animals were euthanized 8 weeks after implantation, and transverse sections of femur-implant samples were used for histology, micro-computed tomography and immunohistochemical evaluations. In group B, the expression of α2ß1 and α5ß1 integrins, which are known to mediate osteoblast adhesion, glycosaminoglycans, heparan sulfate and chondroitin sulfate, was less than half of that in group C. Important to this study, the zirconia surface used in group A prevented these deficiencies. Therefore, these results indicate that nanostructured zirconia surface created on clinical implants by ion beam-assisted deposition may prevent impaired osseointegration in elderly patients by promoting quicker maturation of peri-implant woven bone.


Subject(s)
Femur/physiology , Femur/surgery , Hip Prosthesis , Metal Nanoparticles/chemistry , Osseointegration/physiology , Titanium/chemistry , Zirconium/chemistry , Animals , Coated Materials, Biocompatible/chemistry , Femur/cytology , Male , Metal Nanoparticles/ultrastructure , Particle Size , Rats , Rats, Sprague-Dawley , Surface Properties , Treatment Outcome
2.
Int J Med Microbiol ; 303(3): 105-13, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23474241

ABSTRACT

Live, attenuated Salmonella enterica serovar Typhi strain Ty21a, a licensed oral typhoid fever vaccine, has also been employed for use as a vector to deliver protective antigens of Shigella and other pathogens. Importantly, lipopolysaccharide (LPS) alone has been shown to be a potent antigen for specific protection against shigellosis. We reported previously the plasmid cloning of heterologous LPS biosynthetic genes and the expression in Ty21a of either S. sonnei or of S. dysenteriae 1 LPS's. The resulting plasmids encoding Shigella LPS's were reasonably stable for >50 generations of growth in nonselective media, but still contained an antibiotic resistance marker that is objectionable to vaccine regulatory authorities. Deletion of this antibiotic-resistance marker inexplicably resulted in significant plasmid instability. Thus, we sought a method to insert the large ∼12kb S. sonnei LPS gene region into the chromosome, that would allow for subsequent removal of a selectable marker and would result in 100% genetic stability. Toward this objective, we optimized an existing recombination method to mediate the insertion of a ∼12kb region encoding the S. sonnei LPS genes into the Ty21a genome in a region that is nonfunctional due to mutation. The resulting strain Ty21a-Ss simultaneously expresses both homologous Ty21a and heterologous S. sonnei O-antigens. This chromosomal insert was shown to be 100% genetically stable in vitro and in vivo. Moreover, Ty21a-Ss elicited strong dual anti-LPS serum immune responses and 100% protection in mice against a virulent S. sonnei challenge. This new vaccine candidate, absolutely stable for vaccine manufacture, should provide combined protection against enteric fevers due to Salmonella serovar Typhi as shown previously (and some Paratyphi infections) and against shigellosis due to S. sonnei.


Subject(s)
Bacterial Vaccines/immunology , Drug Carriers/administration & dosage , Drug Delivery Systems , O Antigens/biosynthesis , Salmonella typhi/genetics , Shigella sonnei/immunology , Animals , Antibodies, Bacterial/blood , Bacterial Vaccines/administration & dosage , Biosynthetic Pathways , Chromosomes, Bacterial , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Disease Models, Animal , Dysentery, Bacillary/prevention & control , Female , Genomic Instability , Mice , Mice, Inbred BALB C , Molecular Biology/methods , Molecular Sequence Data , O Antigens/genetics , Plasmids , Sequence Analysis, DNA , Shigella sonnei/genetics , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/immunology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/immunology
3.
Microb Pathog ; 52(2): 140-7, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22023990

ABSTRACT

The mechanism of Cronobacter pathogenesis in neonatal meningitis and potential virulence factors (aside from host cell invasion ability) remain largely unknown. To ascertain whether Cronobacter can invade and transcytose across intestinal epithelial cells, enter into the blood stream and then transcytose across the blood-brain-barrier, we have utilized human intestinal INT407 and Caco-2 cells and brain microvascular endothelial cell (HBMEC) monolayers on Transwell filters as experimental model systems. Our data indicate a wide range of heterogeneity with respect to invasion efficiency among twenty-three Cronobacter isolates screened. For selected isolates, we observed significant levels of transcytosis for Cronobacter sakazakii across tight monolayers of both Caco-2 and HBMEC, mimicking in vivo ability to cross the intestine as well as the blood brain barrier, and at a frequency equivalent to that of a control meningitis-causing Escherichia coli K1 strain. Finally, EM analysis demonstrated intracellular Cronobacter bacteria within host vacuoles in HBMEC, as well as transcytosed bacteria at the basolateral surface. These data reveal that certain Cronobacter isolates can invade and translocate across both cultured human intestinal epithelial cells and HBMEC, thus demonstrating a potential path for neonatal infections of the central nervous system (CNS) following oral ingestion.


Subject(s)
Cronobacter/pathogenicity , Endothelial Cells/microbiology , Epithelial Cells/microbiology , Transcytosis , Cell Line , Cytoplasm/microbiology , Escherichia coli/pathogenicity , Humans , Intestines/cytology , Microscopy, Electron , Vacuoles/microbiology , Virulence
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