Biopolymer-based membranes associated with osteogenic growth peptide for guided bone regeneration.

Barrier membranes for guided bone regeneration (GBR) mainly promote mechanical maintenance of bone defect space and induce osteopromotion. Additionally, biopolymer-based membranes may provide greater bioactivity and biocompatibility due to their similarity to extracellular matrix (ECM). In this study, biopolymers-based membranes from bacterial cellulose (BC) and collagen (COL) associated with osteogenic growth peptide (OGP(10-14)) were evaluated to determine in vitro osteoinductive potential in early osteogenesis; moreover, histological study was performed to evaluate the BC-COL OGP(10-14) membranes on bone healing after GBR in noncritical defects in rat femur. The results showed that the BC-COL and BC-COL OGP(10-14) membranes promoted cell proliferation and alkaline phosphatase activity in osteoblastic cell cultures. However, ECM mineralization was similar between cultures grown on BC OGP(10-14) and BC-COL OGP(10-14) membranes. In vivo results showed that all the membranes tested, including the peptide-free BC membrane, promoted better bone regeneration than control group. Furthermore, the BC-COL OGP(10-14) membranes induced higher radiographic density in the repaired bone than the other groups at 1, 4 and 16 weeks. Histomorphometric analyses revealed that the BC-COL OGP(10-14) induced higher percentage of bone tissue in the repaired area at 2 and 4 weeks than others membranes. In general, these biopolymer-based membranes might be potential candidates for bone regeneration applications.

Rare-earth doped fluoride phosphate glasses: structural foundations of their luminescence properties.

We report a detailed structural investigation of a series of fluoride-phosphate glasses with different phosphate/fluoride ratios in the system xSr(PO3)2-(100 - x)[AlF3-CaF2-SrF2-MgF2] with x = 5, 10, 20, 40. Raman and multinuclear solid NMR spectroscopies confirm that the polyphosphate network structure is successively transformed to a structure dominated by Al-O-P linkages with increasing AlF3 content. Average numbers of Al-O-P linkages have been quantified by 27Al/31P NMR double-resonance techniques. The majority of the fluoride species are found in an alkaline earth metal/aluminum rich environment. The local environments for rare-earth ions have been characterized by EPR spectroscopy of Yb3+ ion spin probes and by photoluminescence experiments on Eu3+ dopant ions, including the 5D0 → 7F2 and 5D0 → 7F1 transition intensity ratio, the normalized phonon sideband intensities in the excitation spectra, and the lifetime of the 5D0 excited state. The results indicate clear correlations between these parameters as a function of composition, and confirm that even at the highest fluoride levels, there is still some residual rare-earth phosphate coordination.

Regenerated cellulose scaffolds: Preparation, characterization and toxicological evaluation.

Regenerated cellulose scaffolds (RCS) may be used as alloplastic materials for tissue repair. In this work, the RCS were obtained by viscose process and characterized by scanning electron microscopy (SEM), wide angle X-ray diffraction (WAXD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetry analysis (TG). In vitro enzymatic degradation assay and toxicological assays were also evaluated. The physicochemical characterizations revealed the formation of a porous material with distinct thermal profile and crystallinity compared to pristine cellulose pulp. Enzymatic degradation assay revealed that lysozyme showed a mildest catalytic action when compared to cellulase, Tricoderma reesei (Tr). Nevertheless, both enzymes were efficient for degrading the RCS. RCS did not show cytotoxicity, mutagenic or genotoxic effects. The systematically characterization of this work suggests that RCS presented distinct features that make it a viable material for future studies related to the development of scaffolds for biological applications.

Carbonic anhydrase XII in valve interstitial cells promotes the regression of calcific aortic valve stenosis.

AIMS: Calcific aortic valve stenosis (CAVS) is the most common heart valve disease. In the present work we sought to determine the reversibility of mineralization in the aortic valve. METHODS AND RESULTS: By using in vitro analyses we found that valve interstitial cells (VICs) have the ability to resorb minerals. We documented that agonist of P2Y2 receptor (P2Y2R) promoted the expression of carbonic anhydrase XII (CAXII) at the cell membrane of VICs, whereby minerals are resorbed. P2Y2R-mediated mineral resorption was corroborated by using mouse VICs isolated from wild type and P2Y2R(-/-) mice. Measurements of extracellular pH (pHe) by using core-shell nanosensors revealed that P2Y2R-mediated CAXII export to the cell membrane led to an acidification of extracellular space, whereby minerals are resorbed. In vivo, we next treated LDLR(-/-)/ApoB(100/100)/IGF2 mice, which had developed CAVS under a high-fat/high-sucrose diet for 8 months, with 2-thioUTP (a P2Y2R agonist) or saline for the next 2 months. The administration of 2-thioUTP (2mg/kg/day i.p.) reduced the mineral volume in the aortic valve measured with serial microCT analyses, which improved hemodynamics and reduced left ventricular hypertrophy (LVH). Examination of leaflets at necropsy confirmed a lower level of mineralization and fibrosis along with higher levels of CAXII in mice under 2-thioUTP. In another series of experiment, the administration of acetazolamide (a CA inhibitor) prevented the acidification of leaflets and the regression of CAVS induced by 2-thioUTP in LDLR(-/-)/ApoB(100/100)/IGF2 mice. CONCLUSION: P2Y2R-mediated expression of CAXII by VICs acidifies the extracellular space and promotes the regression of CAVS.

Production of biocompatible and antimicrobial bacterial cellulose polymers functionalized by RGDC grafting groups and gentamicin.

Bacterial cellulose (BC), a three-dimensional fibril, is a natural polymer that can be used for many applications. BC effectiveness may be improved by enhancing surface characteristics contributing to a better physiologic interaction with human and animal cells and to intrinsically present antimicrobial agents. In the present study, gentamicin-activated BC membranes were obtained by chemically grafting RGDC peptides (R: arginine; G: glycine; D: aspartic acid; C: cysteine) using coupling agent 3-aminopropyltriethoxysilane (APTES) followed by covalent attachment of gentamicin onto the surface of the BC membrane network. X-ray photoelectron spectroscopy (XPS) analyses showed that the BC-APTES contained 0.7% of silicon in terms of elemental composition, corresponding to a grafting ratio of 1:12. The presence of silicon and nitrogen in the BC-APTES confirmed the surface functionalization of the BC membrane. Fourier-transform infrared (FTIR) analyses show the formation of the secondary amide as supported by the valence bond C═O (ν(C═O)), a characteristic vibrational transition at 1650 cm(-1) which is particularly intense with the BC-RGDC-gentamicin membrane. Energy-dispersive X-ray (EDX) analyses showed a low level of carbon and nitrogen (C + N) in pure BC but a high level of (C + N) in BC-RGDC-gentamicin confirming the surface modification of the BC membrane by RGDC and gentamicin enrichment. Of great interest, the gentamicin-RGDC-grafted BC membranes are bactericidal against Streptococcus mutans but nontoxic to human dermal fibroblasts and thus may be useful for multiple applications such as improved wound healing and drug delivery systems.

Mechanical strain induces the production of spheroid mineralized microparticles in the aortic valve through a RhoA/ROCK-dependent mechanism.

Calcific aortic valve disease (CAVD) is a chronic disorder characterized by an abnormal mineralization of the leaflets, which is accelerated in bicuspid aortic valve (BAV). It is suspected that mechanical strain may promote/enhance mineralization of the aortic valve. However, the effect of mechanical strain and the involved pathways during mineralization of the aortic valve remains largely unknown. Valve interstitial cells (VICs) were isolated and studied under strain conditions. Human bicuspid aortic valves were examined as a model relevant to increase mechanical strain. Cyclic strain increased mineralization of VICs by several-fold. Scanning electron microscope (SEM) and energy dispersive X-ray (EDX) analyses revealed that mechanical strain promoted the formation of mineralized spheroid microparticles, which coalesced into larger structure at the surface of apoptotic VICs. Apoptosis and mineralization were closely associated with expression of ENPP1. Inhibition of ENPP1 greatly reduced mineralization of VIC cultures. Through several lines of evidence we showed that mechanical strain promoted the export of ENPP1-containing vesicles to the plasma membrane through a RhoA/ROCK pathway. Studies conducted in human BAV revealed the presence of spheroid mineralized structures along with the expression of ENPP1 in areas of high mechanical strain. Mechanical strain promotes the production and accumulation of spheroid mineralized microparticles by VICs, which may represent one important underlying mechanism involved in aortic valve mineralization. RhoA/ROCK-mediated export of ENPP1 to the plasma membrane promotes strain-induced mineralization of VICs.

Estudo microbiológico comparativo entre sabão de clorexidina 2% e um novo sabão antisséptico para lavagem pré-operatória das mãos / Microbiological study comparing 2% chlorhexidine soap and a new antiseptic soap for preoperative handwashing

Objetivos: avaliar, por meio de estudo microbiológico, dois tipos de sabão propostos para a lavagem das mãos na preparação dos cirurgiões no tempo pré-operatório; comparando um sabão com potência antibacteriana já conhecida e um novo sabão formulado a partir de óleos vegetais. Materiais e métodos: Dez voluntários fizeram a escovação das mãos, segundo protocolos pré-estabelecidos para a rotina de medidas de antissepsia em centros cirúrgicos, com 3 sabões diferentes, sendo um sabão comum (comercial) sem poder antisséptico que serviu como Grupo Controle (I), um sabão de digluconato de clorexidina a 2% (Grupo II) e um novo sabão feito a partir de óleos vegetais numa concentração de 20%, que foi desenvolvido pelo Instituto de Química Unesp/araraquara no Grupo de Materiais Fotônicos, denominado surfactante a 20% (Grupo III). antes de lavar as mãos, logo após e uma hora depois com o uso de luvas cirúrgicas, foi realizada a coleta microbiológica. Resultados: Pelos resultados da aNoVa, verificam-se as seguintes diferenças significativas para o número de colônias bacterianas: entre tipos de sabão (menor número de colônias no sabão do Grupo II), entre tempos (redução do número de colônias no sabão do Grupo II) e efeito significativo da interação sabão versus tempo. Conclusão: o sabão de digluconato de clorexidina 2% mostrou um comportamento melhor em reduzir o número de colônias bacterianas das mãos imediatamente após a lavagem e continuou sendo superior após uma hora com o uso de luvas, quando comparado ao sabão de surfactante a 20%.

Objective: To evaluate, by means of a microbiologic study, two kinds of soaps suggested by surgeons for presurgical handwashing, comparing a well-known antibacterial soap with a new soap formulated from vegetable oils. Materials and methods: Ten volunteers performed handwashing according to previously established protocols for routine antisepsis in operating rooms using 3 different soaps: a common, commercially marketed soap, serving as the control group (Group 1), with no antibacterial characteristics; a soap with 2% chlorhexidine (Group II); and a new soap formulated from vegetable oils at a concentration of 20%, known as surfactant, which was designed by the Chemistry Institute (Unesp/Araraquara - Grupo de Materiais Fotônicos) (Group III). The microbiological samples were collected immediately before and after handwashing and one hour later with the volunteer wearing surgical gloves. Results: ANOVA revealed that the following significant differences are found in the number of bacterial colonies: between soap types (a smaller number of colonies in the Group II soap), between periods (reduction in the number of colonies in the Group II soap), and the significant effect of the soap versus time interaction. Conclusion: The 2% chlorhexidine soap performed better in reducing the number of bacterial colonies on the hands immediately after handwashing and after one hour with the use of surgical gloves, when compared to the 20% surfactant soap.

Determination of olive oil acidity by CE.

In this work, a CE method for the determination of olive oil acidity was proposed. The method was based on an ethanolic extraction (at 60 degrees C) of the oil long-chain free fatty acids (LC-FFAs) components followed by CE determination in pH 6.86 phosphate buffer at 15 mmol/L concentration containing 4 mmol/L sodium dodecylbenzenesulfonate (SDBS), 10 mmol/L polyoxyethylene 23 lauryl ether (Brij 35), 2% v/v 1-octanol and 45% v/v ACN under indirect UV detection at 224 nm. Although this electrolyte promoted baseline separation of myristic acid (C14:0) (internal standard (IS)) and olive oil major components (palmitic acid (C16:0), oleic acid (C18:1c) and linoleic acid (C18:2cc)) in less than 8 min, after a few injections, the electropherogram profiles were severely altered (peak broadening, migration time shifts, etc.) and the current increased substantially. An adsorption study was conducted revealing that the dissolution of the capillary external polyimide coating during the electrophoretic run caused the detrimental effect. After removal of the capillary tip coating, ten consecutive injections could be performed without any disturbances and this simple procedure was, therefore, implemented during quantitative purposes. The reliability of the proposed method was further investigated by the determination of acidity of an extra virgin olive oil sample in comparison to the established methodology (AOCS method Ca 5a-40, alkaline volumetric titration (AVT)). No statistical differences were found within 95% confidence level. A % acidity of 0.39 +/- 0.02 was found for the olive oil sample under consideration.

Analysis of sphingosine 1-phosphate by capillary electrophoresis coupled to laser-induced fluorescence detection: use of a transparent fused-silica capillary.

C18-sphingosine 1-phosphate (C18-SPP) is a sphingolipid with important functional and structural roles in cells. In this paper we report a new capillary electrophoresis technique that is coupled to a laser-induced fluorescence detection (CE-LIF) method to quantify the level of C18-SPP in biological samples. The method utilizes a commercial standard C17-sphingosine 1-phosphate (C17-SPP) derivatized with naphthalene-2,3-dicarboxaldehyde (NDA), a fluorogenic dye used to label primary amines. The detection limit for the C17-SPP NDA was 0.54 fmol. We quantified the C18-SPP in leukemic human cells before and after irradiation by gamma rays. We demonstrated that the amounts of this sphingolipid decreased after the irradiation. In a second part of this work, we used the technique to evaluate the ability of a novel transparent fused-silica capillary, which allows the use of fused-silica capillaries without burning a window. Solarization, homogeneity, and sensitivity were studied using this capillary. The results demonstrate that this new durable capillary can provide a sensitive and reproducible quantitation procedure for CE-LIF studies.

Spectroscopic investigation of a new hybrid glass formed by the interaction between croconate ion and calcium polyphosphate.

In this work, a new organic-inorganic hybrid material has been synthesized by the incorporation of croconate ion into a calcium polyphosphate coacervate. The hybrid so obtained was characterized by means of electronic and vibrational spectroscopies. The material is a homogeneous mixture described by a structural model, which includes helical chains of polyphosphate ions, where the calcium ion occupies the internal vacancies of the structure. The croconate ion appears to be occupying the regions outside the polymeric structure, surrounded by several water molecules. The electronic spectrum of the incorporated material shows a broad band peaking at the same wavelength region (363 nm) observed for the aqueous solution of croconate ion, and manifesting the Jahn-Teller effect as evidenced by the doublet structure of the band. The infrared spectrum is widely dominated by the absorption bands of the polyphosphate ion and the appearance of the carbonyl stretching band at ca. 1550 cm(-1) indicates the presence of croconate ion incorporated in the structure. The Raman spectrum of the material shows several vibrational bands related to the oxocarbon moiety; most of them are shifted in comparison with the free ion. These shifts can be understood in terms of strong hydrogen bonding interactions between water molecules and the oxocarbon moiety. The low temperature methodology proposed here can be well used in the preparation of new phosphate glasses containing organic moieties opening the route to an entirely new class of hybrid glasses.