Dentin Priming with Amphipathic Antimicrobial Peptides.

The evolution of bonded restorations has undergone great progress over several decades. Nonetheless, life spans of bonded restorations are limited mainly because of the eventual incidence of recurrent caries. Over time, water and waterborne agents (acids, enzymes) degrade the components of the dentin/restoration interface, allowing bacterial colonization and dentin reinfection at the margins of the restoration. We developed a 2-tier protective technology consisting of priming/coating dentin with amphipathic and antimicrobial peptides (AAMPs) to obtain hydrophobic/water-repellent and antibiofilm dentin-resisting recurrent caries around bonded restorations. We tested a series of AAMPs to assess their structure-function relationships as well as the effects of different dentin-conditioning methods on the structural features of AAMP-coated dentin. We found relation between the secondary structure of AAMPs (high portion of ß-sheet), the antimicrobial potency of AAMPs, and the AAMPs' ability to form hydrophobic coatings on dentin. We also determined that AAMPs had preferential adsorption on the mineral phase of dentin, which suggested that peptides arrange their cationic and hydrophilic motifs in direct contact with the negatively charged minerals in the hydrophilic dentin. These results led us to explore different dentin-conditioning methods that would increase the mineral/collagen ratio and their effect on AAMP immobilization. We innovatively imaged the spatial distribution of the AAMPs in relation to the dentinal tubules and collagen network using a minimally invasive multimodal imaging technique: multiphoton-second harmonic generation. Using multiphoton-second harmonic generation imaging, we determined that partial deproteinization of dentin increased the amount of immobilized AAMPs as compared with the total etched dentin at the dentin surface and extended deeply around dentinal tubules. Last, we analyzed the release rate of AAMPs from dentin coatings in artificial saliva to predict their stability in the clinical setting. In conclusion, priming dentin with AAMPs is a versatile new approach with potential to fortify the otherwise vulnerable adhesive-based interfaces.

Studies on thermostable antigens, production of species-specific antiadrenal sera and comparison of immunological techniques in meat speciation.

Heat-stable antigens (BE forms: resistant to heat and ethanol precipitation) of adrenal and muscle tissues of cattle, buffalo, sheep, goat and pig were prepared for use in detection of adulteration in meats. The physico-chemical characteristics of these antigens revealed that the antigens of adrenals had only one component corresponding to 'Troponin T'. Muscle antigens also contained a major troponin T component but were associated with low molecular weight fractions. Rabbit antiadrenal BE sera were developed and made species specific by immunoabsorption. The species-specific antisera were employed for identification of origin of fresh and cooked meats and their mixtures, using an immunodiffusion test-agar gel precipitation test (AGPT), counterimmunoelectrophoresis (CIEP), enzyme-linked immunosorbent assay (ELISA) and the unlabelled antibody peroxidase antiperoxidase (PAP) technique. The results indicated that absorbed antisera could successfully differentiate the fresh, cooked meats and the meat mixtures from the species under study. AGPT and CIEP were useful in identification of 5-10% addition, using water extracts of fresh meats and BE forms of cooked meats, whereas ELISA and PAP could detect adulteration down to the level of 1% when water extracts were used. Among the tests employed in the study, the PAP technique proved to be most sensitive. The antisera were also proved useful in identifying the species in canned meat products, milk, serum, plasma, semen, urine, organs, skin and spoilt flesh, employing AGPT and CIEP.