On the prevalence of flow instabilities from high-fidelity computational fluid dynamics of intracranial bifurcation aneurysms.

High-fidelity computational fluid dynamics (HF-CFD) has revealed the potential for high-frequency flow instabilities (aka "turbulent-like" flow) in intracranial aneurysms, consistent with classic in vivo and in vitro reports of bruits and/or wall vibrations. However, HF-CFD has typically been performed on limited numbers of cases, often with unphysiological inflow conditions or focused on sidewall-type aneurysms where flow instabilities may be inherently less prevalent. Here we report HF-CFD of 50 bifurcation aneurysm cases from the open-source Aneurisk model repository. These were meshed using quadratic finite elements having an average effective spatial resolution of 0.065 mm, and solved under physiologically-pulsatile flow conditions using a well-validated, minimally-dissipative solver with 20,000 time-steps per cardiac cycle Flow instability was quantified using the recently introduced spectral power index (SPI), which quantifies, from 0 to 1, the power associated with velocity fluctuations above those of the driving inflow waveform. Of the 50 cases, nearly half showed regions within the sac having SPI up to 0.5, often with non-negligible power into the 100's of Hz, and roughly 1/3 had sac-averaged SPI > 0.1. High SPI did not significantly predict rupture status in this cohort. Proper orthogonal decomposition of cases with highest SPIavg revealed time-varying energetics consistent with transient turbulence. Our reported prevalence of high-frequency flow instabilities in HF-CFD modelling of aneurysms suggests that care must be taken to avoid routinely overlooking them if we are to understand the highly dynamic mechanical forces to which some aneurysm walls may be exposed, and their prevalence in vivo.

Osteoblast-like cell adhesion to bone sialoprotein peptides.

A number of studies have suggested that biomimetic peptides can be used in the design of a new generation of prosthetic implants to promote the successful biointegration of the implant materials. In the current study, the in vitro bioactivities of several peptides representing RGD (Arg-Gly-Asp)-containing sequences of bone sialoprotein (BSP) toward an osteoblast-like cell line (MC3T3-E1) were examined to provide insight into the molecular basis of BSP's interaction with bone cells. BSP residues 283-288, 281-290, 278-293 and 278-302 were coated on polystyrene surfaces in 96-well non-tissue (untreated) culture plates, and their osteoblast adhesive properties compared to intact BSP and fibronectin as positive controls. BSP peptides 278-302 and 278-293 were found to be the most potent in their adhesive activity, increasing the number of adherent cells to 350% of control levels at an added concentration of 1 microM. Since these two peptides were equivalent in potency, it is suggested that the region 294-302 beyond the RGD domain is not necessary for cell binding. In comparison, peptides 283-288 and 281-290 were only active at concentrations greater than 200 microM. 50-70% of the peptide-stimulated adhesion was inhibited by the pretreatment of cell suspensions with solution phase RGD, suggesting that a portion of the peptides' adhesive effects was specific and integrin-mediated, although other non-RGD flanking regions were probably also involved in the mechanism of adhesion. Importantly, a modified BSP peptide, in which an aspartic acid residue at position 288 of the RGD sequence was replaced by a glutamic acid residue to form RGE, was completely inactive as a cell adhesion stimulus at concentrations up to 200 microM. Thus, despite the potential role of non-RGD flanking regions, an intact RGD tripeptide was essential for all of the adhesive activity of the BSP peptides.

Thermal and chemical modification of titanium-aluminum-vanadium implant materials: effects on surface properties, glycoprotein adsorption, and MG63 cell attachment.

The microstructure, chemical composition and wettability of thermally and chemically modified Ti-6Al-4V alloy disks were characterized and correlated with the degree of radiolabeled fibronectin-alloy surface adsorption and subsequent adhesion of osteoblast-like cells. Heating either in pure oxygen or atmosphere (atm) resulted in an enrichment of Al and V within the surface oxide. Heating (oxygen/atm) and peroxide treatment both followed by butanol treatment resulted in a reduction in content of V, but not in Al. Heating (oxygen/atm) or peroxide treatment resulted in a thicker oxide layer and a more hydrophilic surface when compared with passivated controls. Post-treatment with butanol, however, resulted in less hydrophilic surfaces than heating or peroxide treatment alone. The greatest increases in the adsorption of radiolabeled fibronectin following treatment were observed with peroxide/butanol-treated samples followed by peroxide/butanol and heat/butanol, although binding was only increased by 20-40% compared to untreated controls. These experiments with radiolabeled fibronectin indicate that enhanced adsorption of the glycoprotein was more highly correlated with changes in chemical composition, reflected in a reduction in V content and decrease in the V/Al ratio, than with changes in wettability. Despite promoting only a modest elevation in fibronectin adsorption, the treatment of disks with heat or heat/butanol induced a several-fold increase in the attachment of MG63 cells promoted by a nonadhesive concentration of fibronectin that was used to coat the pretreated disks compared to uncoated disks. Therefore, results obtained with these modifications of surface properties indicate that an increase in the absolute content of Al and/or V (heat), and/or in the Al/V ratio (with little change in hydrophilicity; heat+butanol) is correlated with an increase in the fibronectin-promoted adhesion of an osteoblast-like cell line. It would also appear that the thermal treatment-induced enhancement of cell adhesion in the presence of this integrin-binding protein is due to its increased biological activity, rather than a mass effect alone, that appear to be associated with changes in chemical composition of the metallic surface. Future studies will investigate the influence of the surface chemical composition of various implantable alloys on protein adsorption and receptor-mediated cell adhesion. In addition, by altering the properties of bound osteogenic protein enhancing exposure to cell integrin binding domains, it may be possible to develop implant surfaces which enhance the attachment, adhesion and developmental response of osteoblast precursors leading to accelerated osseointegration.

Adsorption and dissolution behavior of human plasma fibronectin on thermally and chemically modified titanium dioxide particles.

Titanium is known for its biocompatibility and is widely used in dental and orthopedic reconstructive surgery. There are reports that osteointegration of these implants is not optimal. The objective of this study was to modify titanium dioxide particles and examine the resultant effects on protein adsorption to these altered surfaces using a model cell binding protein, human plasma fibronectin (HPF). HPF is an important matrix glycoprotein that plays a major role in cell and protein attachment, Titanium dioxide surfaces were modified by heating the titanium dioxide powder at 800 degrees C for 1 h or treating with an oxidizing agent: peroxide in ammonium hydroxide followed by peroxide in hydrochloric acid. Oxidized and control samples were further treated with 9:1 butanol:water for 30 min. Brunauer-Emmett-Teller showed no change in particle surface area as a result of thermal or chemical treatment. Hydrophobicity increased with butanol treatment of titanium dioxide. Diffuse reflectance Fourier transform infrared spectroscopy showed the presence of -CH2 and -CH3 vibrations in the region of 2850-3000 cm(-1) for both the heated, butanol and peroxide/butanol-treated samples. The absence of increased C-O and O-C=O features as determined by electron spectroscopy for chemical analysis indicates that butanol adsorption is not occurring via an esterification mechanism. The interaction between butanol and pre-heated or peroxide-treated titanium dioxide may be one of association (weak electrostatic and/or Van der Waals forces) rather than direct ionic bonding. Maximum HPF adsorption on modified or unmodified titanium dioxide occurred within 30 min, with greater protein adsorption occurring on butanol-treated samples. Desorption was minimal with all modifications. Zeta potential measurements showed that HPF adsorption caused an increase in the negative zeta potential with the greatest change noted for the butanol-treated samples. These findings suggest that wettability and surface charge both play an important role in protein adsorption to titanium dioxide. Thus, by modifying the physico-chemical properties of titanium dioxide surfaces, it may be possible to alter protein adsorption and hence optimize cell attachment.

Physicochemical study of plasma-sprayed hydroxyapatite-coated implants in humans.

This study represents the first report of the physical and chemical changes occurring in coatings of failed hydroxyapatite (HA)-coated titanium implants obtained from a comprehensive, multicenter human dental implant study. A total of 53 retrieved samples were obtained and compared with unimplanted controls with the same manufacturer and similar manufacture dates. Forty-five retrieved implants were examined for surface characteristics and bulk composition. Implants were staged based on implantation history: stage 1 (implants retrieved between surgical placement and surgical uncovering), stage 2 (implants retrieved at surgical uncovering and evaluation), stage 3 (implants retrieved between surgical uncovering evaluation and occlusal loading), and stage 4 (implants retrieved after occlusal loading). Scanning electron microscopy showed progressive coating thinning with implantation time. At later stages, bare Ti metal was detected by energy-dispersive X-ray analysis and electron spectroscopy for chemical analysis. Increases in Ti and Al (2-7.5 atm % each) were detected at the apical ends of all stage 4 samples. In unimplanted coatings, X-ray diffraction analysis demonstrated the presence of amorphous calcium phosphate, beta-tricalcium phosphate, tetracalcium phosphate, and calcium oxide in addition to large hydroxyapatite crystals (c axis size, D002 = 429 +/- 13 A; a axis size, D300 = 402 +/- 11 A, a/c aspect ratio 0.92). The nonapatitic phases disappeared with increased implantation time, although there was a persistence of amorphous calcium phosphate. Bulk coating chemical analysis showed that Ca/P ratios for implant controls (1.81 +/- 0.01) were greater than stoichiometric HA (1.67) and decreased for implant stages 3 and 4 (1.69 +/- 0.09 and 1.67 +/- 0.09, respectively), explained by the dissolution of the non apatitic phases. Crystal sizes also changed with implantation times, being smaller than the control at all but stage 4. Fourier transform infrared analyses agreed with these results, and also indicated the accumulation of bone (protein and carbonate-apatite) in the retrieved coatings. The accumulation of bone was not stage dependent. These findings indicate that there was some biointegration with the surrounding bone, but the greatest changes occurred with the HA coating materials, their loss, and chemical change.

Effect of evaporation and mixing technique on die spacer thickness: a preliminary study.

STATEMENT OF PROBLEM: Casting relief is required for proper seating of castings to allow for luting agent thickness. The application of die spacer to the die is the most common method of obtaining casting relief. Die spacer film thicknesses that are outside the ideal range of 25 to 40 microm can cause clinical problems. Thickness can be affected by the separation of die spacer constituents, which may not be reconstituted by mixing, in the bottle and by the evaporation of volatile components while the bottle is open. PURPOSE: The purpose of this study was to determine the effects of component evaporation and die spacer mixing technique on applied die spacer thickness. MATERIAL AND METHODS: Bottles of Gold Tru-fit die spacer were left open for 0, 1, 4, 8, and 24 hours at 22 degrees C. Spacer solutions were shaken either by hand per the manufacturer's directions or on a dental vibrator for 1 minute. One even brush stroke of spacer was applied to clean glass slides. Three die spacer films were made for each combination of time and mixing technique. Eighteen thickness measurements per sample at various sites were recorded with profilometer tracings. Statistical differences were determined with a 2-way ANOVA. RESULTS: Handshaking provided greater die spacer thickness, which increased with the time that the bottle was open. Vibration provided lower thickness with no statistical increase with time. CONCLUSION: Insufficient agitation caused lower film thickness. Excessive evaporation caused higher film thickness.

A methodological study for the analysis of apatite-coated dental implants retrieved from humans.

The stability of thermally processed hydroxyapatite coatings for oral and orthopedic bioprostheses has been questioned. Information on the chemical changes, which occur with hydroxyapatite biomaterials post-implantation in humans, is lacking. The purpose of this investigation was to begin to examine post-implantation surface changes of hydroxyapatite-coated implants using scanning electron microscopy (SEM), x-ray microanalysis (EDAX), Fourier transform infrared spectroscopy (FTIR), and x-ray diffraction (XRD). Three retrieved dental implant specimens from humans following clinical failure due to peri-implantitis were examined. Unimplanted cylinders served as controls. Clinically, the retrieved specimens were all enveloped by a fibrous tissue capsule with bone present at the apical extent of the implant. SEM analysis showed that the retrieved surfaces were coated with both calcified and proteinaceous deposits. EDAX scans of the retrieved specimens demonstrated evidence of hydroxyapatite coating loss reflected by increasing titanium and aluminum signals. Other foreign ions such as sodium, chloride, sulfur, silica, and magnesium were detected. XRD of the control specimens showed that the samples were predominantly apatite; however, two peaks were detected in the diffraction pattern, which are not characteristic of hydroxyapatite, indicating that small amounts of one or more other crystalline phases were also present. The retrieved specimens showed slightly larger average crystal size relative to the control sample material, and the non-apatite lines were not present. FTIR evaluation of the retrieved specimens revealed the incorporation of carbonate and organic matrix on or into the hydroxyapatite. Narrowing of and increased detail in the phosphate peaks indicated an increase in average crystal size and/or perfection relative to the controls, as did the XRD results. Based on these results, we conclude that chemical changes may occur within the coating, with the incorporation of carbonate and concomitant reduction in hydroxyapatite coating thickness. Thermodynamic dissolution-reprecipitation of the coating itself and subsequent surface insult by bacterial and local inflammatory components may be involved with these changes.

Surface analysis of human plasma fibronectin adsorbed to commercially pure titanium materials.

Protein binding on metallic implant surfaces, such as titanium, is governed by the physico-chemical nature of the metallic surface. Human plasma fibronectin (HPF) is an important matrix glycoprotein that mediates cell and protein attachment to each other or to the extracellular matrix present during wound healing. The objective of this study was to investigate the adsorption of HPF onto polished commercially pure titanium (cpTi) by using atomic force microscopy (AFM) and electron spectroscopy for chemical analysis (ESCA) and to measure the resultant surface contact angle before and after HPF binding. Two types of cpTi disks, one highly polished in our laboratory (HSS) and one commercially prepared (31), were reacted with HPF solutions of varying concentrations (1 microg/mL-10 ng/mL). ESCA survey spectra of samples coated with 1 microg/mL of fibronectin showed an increase in organic nitrogen and carbon compared with uncoated controls. Contact angle measurements of HSS and 31 cpTi disks showed no significant difference in average contact angle (36.3 degrees +/- 3.5 and 39.1 degrees +/- 3.1) despite differences in local root mean square (RMS) surface roughness (4.45 +/- 0.46 nm and 22.37 +/- 4.17 nm) as measured by AFM. Images obtained by AFM showed that 31 specimens were more irregular, with large parallel polishing grooves. Adsorbed HPF appeared in a globular form with an average length of 16.5 +/- 1.0 nm, a height of 2.5 +/- 0.5 nm, and a width of 9.6 +/- 1.2 nm. Fibronectin coating on both HSS and 31 cpTi specimens resulted in a significant increase in hydrophobicity compared to uncoated specimens. These results indicate the significance of HPF on cpTi and may explain how cpTi implants function in situ.

Chaser (Csr), a new gene affecting larval foraging behavior in Drosophila melanogaster.

Chaser (Csr) was uncovered in a gamma mutagenesis screen to identify genes that modify the larval foraging behavior of sitters to rovers. Rover larvae have significantly longer path lengths than sitters while foraging on a yeast and water paste. This difference is influenced by one major gene, foraging (for), which has two naturally occurring alleles, forR (rover) and fors (sitter). In a mutagenesis screen for modifiers of for, we identified three lines with viable mutations on chromosome 3 that alter foraging behavior. Each of these mutations increased larval path lengths in fors/fors larvae in a dominant fashion, and were not separable by recombination. These mutations are therefore probably allelic and define a new gene that we have called Csr. Csr was genetically localized using the lethal-tagging technique. This technique resulted in seven lines with a significant decrease in larval path-length and recessive lethal mutations on chromosome 3. We refer to these as reverted Csr (Csrrv) lines. Deficiencies that uncovered cytologically visible chromosome rearrangements in three of the seven reverted lines were used in a complementation analysis. In this way we mapped the lethal mutations in the Csrrv lines to cytological region 95F7-96A1 on the right arm of chromosome 3.

Outpatient anesthesia.

More complex methods of outpatient anesthesia, including I.V. sedation and general anesthesia, have become commonplace. Patient selection and preoperative evaluation are discussed, as well as the choice of who will deliver anesthesia. Appropriate outpatient facilities and monitoring are reviewed. The authors' favorite methods of anesthesia, caveats, and suggestions are presented, as well as prevention and treatment of anesthesia problems and emergencies.