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PubMed1994

Diaphragm and lung afferents contribute to inspiratory load compensation in awake ponies.

We determined the effect of pulmonary vagal (hilar nerve) denervation (HND) and diaphragm deafferentation (DD) on inspiratory load compensation. We studied awake intact (I; n = 10), DD (n = 5), HND (n = 4), and DD+HND (n = 7) ponies at rest and during mild (1.8 mph, 5% grade) and moderate (1.8 mph, 15% grade) treadmill exercise before, during, and after resistance of the inspiratory circuit was increased from approximately 1.5 to approximately 20 cmH2O.l-1.s. During the first loaded breath in I ponies at rest, inspiratory time (TI) increased, expiratory time decreased, and inspiratory drive increased. There were minimal changes after the first breath, and inspiratory minute ventilation (VI) and arterial PCO2 did not change (P > 0.10) from control values. On the first loaded breath during exercise, TI increased but inspiratory drive either did not change or decreased from control values. TI and drive increased after the first breath, but the increases were insufficient to maintain VI and arterial PCO2 at control levels. First-breath load compensation remained after DD, HND, and DD+HND, but after DD+HND tidal volume and VI were compensated 5-10% less (P < 0.05) than in I ponies. In all groups inspiratory drive, tidal volume, and VI were markedly augmented on the first breath after loading was terminated with a gradual return toward control. We conclude that diaphragm and pulmonary afferents contribute to but are not essential for inspiratory load compensation in awake ponies.

Animals↗

PubMed2006

In vitro anti-bacterial and biological properties of magnetron co-sputtered silver-containing hydroxyapatite coating.

Bacterial infection after implant placement is a significant rising complication. In order to reduce the incidence of implant-associated infections, several biomaterial surface treatments have been proposed. In this study, the effect of in vitro antibacterial activity and in vitro cytotoxicity of co-sputtered silver (Ag)-containing hydroxyapatite (HA) coating was evaluated. Deposition was achieved by a concurrent supply of 10 W to the Ag target and 300 W to the HA target. Heat treatment at 400 degrees C for 4 h was performed after 3 h deposition. X-ray diffraction, contact angles measurements, and surface roughness were used to characterize the coating surfaces. The RP12 strain of Staphylococcus epidermidis (ATCC 35984) and the Cowan I strain of Staphylococcus aureus were used to evaluate the antibacterial activity of the Ag-HA coatings, whereas human embryonic palatal mesenchyme cells, an osteoblast precursor cell line, were used to evaluate the in vitro cytotoxicity of the coatings. X-ray diffraction analysis performed in this study indicated peaks corresponding to Ag and HA on the co-sputtered Ag-HA surfaces. The contact angles for HA and Ag-HA surfaces were observed to be significantly lower when compared to Ti surfaces, whereas no significant difference in surface roughness was observed for all groups. In vitro bacterial adhesion study indicated a significantly reduced number of S. epidermidis and S. aureus on Ag-HA surface when compared to titanium (Ti) and HA surfaces. In addition, no significant difference in the in vitro cytotoxicty was observed between HA and Ag-HA surfaces. Overall, it was concluded that the creation of a multifunctional surface can be achieved by co-sputtering the osteoconductive HA with antibacterial Ag.

Anti-Bacterial Agents↗

PubMed2003

TEM and STEM analysis on heat-treated and in vitro plasma-sprayed hydroxyapatite/Ti-6Al-4V composite coatings.

A cogent understanding of the microstructure, and indeed nano-structure, of hydroxyapatite (HA) and the interface between Ti-6Al-4V and HA is crucial to its appropriateness as a biomaterials. This paper reports the analysis of plasma-sprayed HA/Ti-6Al-4V composites by transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) to elucidate the intricate nature of the materials following plasma spray processing and in vitro evaluation. The novel Ti-6Al-4V/HA composite coating, with approximately 48 wt% HA, had demonstrated attractive tensile adhesion strength (approximately 28 MPa) and improved Young's modulus (approximately 55 GPa). Experimental results demonstrated that amorphous calcium phosphate and fine HA grains were formed during rapid splat solidification in the as-sprayed composite coatings. Small Ti-6Al-4V grains were observed adjacent to the amorphous calcium phosphate. The coatings were further heat treated at 600 degrees C for 6 h, and significant crystallisation of the amorphous calcium phosphate phase took place. However, complete crystallisation was not achieved at this temperature, as the coatings invariably contained a small amount of amorphous calcium phosphate phase in some local regions. After immersion in simulated body fluid for 2 weeks and 10 weeks, TEM and STEM confirmed that the interfaces inside the coating maintained good microstructural integrity.

Alloys↗

PubMed1978

Hemodynamic studies after De Vega's tricuspid annuloplasty.

Postoperative hemodynamic studies were conducted at rest and during exercise in 24 patients who, in addition to mitral and/or aortic valve surgery, had De Vega's tricuspid annuloplasty. To determine the degree of tricuspid insufficiency (TI), right atrial pressure (PRA) tracings, biplane right ventricular cineangiograms, and ultrasonic Doppler flow patterns were obtained. The TI improved in 14 patients, but still remained mild to moderate in 20 patients. In addition, a mild-to-moderate tricuspid stenosis with pressure gradients from 2 to 7.8 mm Hg were found in 12 patients. There was no significant change in the mean PRA, with 8.0 +/- 4.5 mm Hg preoperatively and 7.5 +/- 3.5 mm Hg postoperatively at rest. During exercise, PRA rose to 17.0 +/- 6.5 mm Hg. The unsatisfactory results are also due to the persistent elevation of left atrial and pulmonary artery pressure after left heart surgery. De Vega's annuloplasty does not answer the tricuspid challenge, since the results are unpredictable.

Adolescent↗

PubMed2007

Plasma-controlled nanocrystallinity and phase composition of TiO2: a smart way to enhance biomimetic response.

This contribution sheds light on the role of crystal size and phase composition in inducing biomimetic apatite growth on the surface of nanostructured titania films synthesized by reactive magnetron sputtering of Ti targets in Ar+O(2) plasmas. Unlike most existing techniques, this method enables one to deposit highly crystalline titania films with a wide range of phase composition and nanocrystal size, without any substrate heating or postannealing. Moreover, by using this dry plasma-based method one can avoid surface hydroxylation at the deposition stage, almost inevitable in wet chemical processes. Results of this work show that high phase purity and optimum crystal size appear to be the essential requirement for efficient apatite formation on magnetron plasma-fabricated bioactive titania coatings.

Apatites↗

PubMed1995

Effect of titanium surface roughness on proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63).

The effect of surface roughness on osteoblast proliferation, differentiation, and protein synthesis was examined. Human osteoblast-like cells (MG63) were cultured on titanium (Ti) disks that had been prepared by one of five different treatment regimens. All disks were pretreated with hydrofluroic acid-nitric acid and washed (PT). PT disks were also: washed, and then electropolished (EP); fine sandblasted, etched with HCl and H2SO4, and washed (FA); coarse sandblasted, etched with HCl and H2SO4, and washed (CA); or Ti plasma-sprayed (TPS). Standard tissue culture plastic was used as a control. Surface topography and profile were evaluated by brightfield and darkfield microscopy, cold field emission scanning electron microscopy, and laser confocal microscopy, while chemical composition was mapped using energy dispersion X-ray analysis and elemental distribution determined using Auger electron spectroscopy. The effect of surface roughness on the cells was evaluated by measuring cell number, [3H]thymidine incorporation into DNA, alkaline phosphatase specific activity, [3H]uridine incorporation into RNA, [3H]proline incorporation into collagenase digestible protein (CDP) and noncollagenase-digestible protein (NCP), and [35S]sulfate incorporation into proteoglycan. Based on surface analysis, the five different Ti surfaces were ranked in order of smoothest to roughest: EP, PT, FA, CA, and TPS. A TiO2 layer was found on all surfaces that ranged in thickness from 100 A in the smoothest group to 300 A in the roughest. When compared to confluent cultures of cells on plastic, the number of cells was reduced on the TPS surfaces and increased on the EP surfaces, while the number of cells on the other surfaces was equivalent to plastic. [3H]Thymidine incorporation was inversely related to surface roughness. Alkaline phosphatase specific activity in isolated cells was found to decrease with increasing surface roughness, except for those cells cultured on CA. In contrast, enzyme activity in the cell layer was only decreased in cultures grown on FA- and TPS-treated surfaces. A direct correlation between surface roughness and RNA and CDP production was found. Surface roughness had no apparent effect on NCP production. Proteoglycan synthesis by the cells was inhibited on all the surfaces studied, with the largest inhibition observed in the CA and EP groups. These results demonstrate that surface roughness alters osteoblast proliferation, differentiation, and matrix production in vitro. The results also suggest that implant surface roughness may play a role in determining phenotypic expression of cells in vivo.

Alkaline Phosphatase↗

PubMed2003

Calcium phosphate coatings obtained by Nd:YAG laser cladding: physicochemical and biologic properties.

The plasma spray (PS) technique is the most popular method commercially in use to produce calcium phosphate (CaP) coatings to promote fixation and osteointegration of the cementless prosthesis. Nevertheless, PS has some disadvantages, such as the poor coating-to-substrate adhesion, low mechanical strength, and brittleness of the coating. In order to overcome the drawbacks of plasma spraying, we introduce in this work a new method to apply a CaP coating on a Ti alloy using a well-known technique in the metallurgical field: laser surface cladding. The physicochemical characterization of the coatings has been carried out by means of X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDX). The biologic properties of the coatings have been assessed in vitro with human osteoblast-like MG-63 cells. The overall results of this study affirm that the Nd:YAG laser cladding technique is a promising method in the biomedical field.

Alloys↗

PubMed2004

Fatigue characteristics of bioactive glass-ceramic-coated Ti-29Nb-13Ta-4.6Zr for biomedical application.

A new surface-coating method by which CaP invert glass is used to improve the bioactivity of titanium alloys has been developed recently. In this method, the powder of CaP invert glass (CaO-P2O5-TiO2-Na2O) is coated on the surface of titanium alloy samples and heated between 1073 and 1123 K. With this treatment, a calcium phosphate layer mainly containing beta-Ca3(PO4)2 phase can be coated easily on titanium alloy samples. In the present study, the effect of this coating process on the fatigue properties of Ti-29Nb-13Ta-4.6Zr, a new metastable beta alloy for biomedical applications, has been investigated. The fatigue endurance limit of the coated alloy was found to be about 15% higher than that of uncoated alloy, as a result of the formation of a hard (alpha + beta) layer and a small amount of the omega phase during the coating process. The coating exhibits excellent adhesion to the substrate during the tensile and fatigue tests. Subsequent ageing at 673 K for 259.2 ks greatly improves the fatigue resistance of the coated alloy due to isothermal omega phase precipitation, and does not have obvious detrimental effect on the coating properties.

Adhesiveness↗

PubMed2006

Laser-deposited Ti-Nb-Zr-Ta orthopedic alloys.

The complex quaternary Ti-35Nb-7Zr-5Ta orthopedic alloy has been successfully deposited from a powder feedstock consisting of a blend of elemental titanium, niobium, zirconium, and tantalum powders, using the laser engineered net-shaping (LENStrade mark) process. In the as laser-deposited form, these alloys exhibit a substantially higher tensile strength as compared with more conventionally processed counterparts of similar composition, while maintaining excellent ductility and a low modulus. Furthermore, the as-deposited alloys appear to exhibit a <001> texture, with a substantially large number of grains of the beta phase aligning one of their <001> axes nearly normal to the substrate or parallel to the growth direction. The microstructure of the as-deposited as well as tensile-tested alloys have been characterized in detail using scanning electron microscopy (SEM), orientation microscopy (OM), and transmission electron microscopy (TEM). Formation of a high density of shear bands, possibly arising from slip localization due to precipitates of the omega phase in the beta matrix, is clearly evident in the tensile-tested sample. The enhanced tensile strength and low modulus in these laser-deposited alloys coupled with the ability to form near-net shape components makes LENS an attractive processing technology for orthopedic implants.

Alloys↗

PubMed1998

Prostaglandins mediate the effects of titanium surface roughness on MG63 osteoblast-like cells and alter cell responsiveness to 1 alpha,25-(OH)2D3.

Surface roughness affects proliferation, differentiation (alkaline phosphatase and osteocalcin), local factor production (transforming growth factor (TGF beta) and prostaglandin E2 (PGE2)], and response to 1,25-(OH)2D3 (1,25) of MG63 osteoblast-like cells. In this study, we examined whether the effect of surface roughness on MG63 cells is mediated by prostaglandins produced by the cells. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then machined and acid-etched (MA). Disks were also coarse grit-sandblasted (SB), coarse grit-sandblasted and acid-etched (CA), or plasma-sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on the Ti disks in the presence or absence of 10(-7) M indomethacin (Indo), a specific inhibitor of cyclooxygenase activity, resulting in decreased prostaglandin production. When the cells reached confluence, cell number, cell layer alkaline phosphatase specific activity (ALPase), and osteocalcin (OC) and latent TGF beta (LTGF beta) production were determined. In addition, confluent cultures which had been grown in the absence of Indo were exposed to 10(-7) M 1,25, 10(-7) M Indo, or a combination of the two for 24 h. On the rougher surfaces, cell number was decreased and ALPase, OC, and LTGF beta were increased. When indomethacin was present throughout the culture period, the effect of surface roughness on cell number, OC, and LTGF beta was abolished. ALPase was reduced, but surface roughness-dependent effects were still observed. Addition of indomethacin to confluent cultures for 24 h had no effect on any of the parameters examined, with one exception: Cells cultured on MA surfaces exhibited a more differentiated phenotype. 1,25 increased all parameters examined on SB, CA, and PS surfaces. When indomethacin was added with 1,25, the 1,25-dependent effects on cell number and OC and LTGF beta production were abolished; however, ALPase was unaffected. This indicates that bone cell response to systemic hormones may be modified by implant surface roughness. This effect may be mediated, at least in part, by prostaglandins produced by the same cells.

Alkaline Phosphatase↗

PubMed2006

[Application of two-photon excitation fluorescence imaging to real-time investigation of mouse preimplantation embryo].

The blue shift of a two-photon excitation absorption peak allows the application of a single wavelength to the simultaneous excitation of several fluorochromes with disparate emission characteristics. An output at 730 nm of a mode-locked femtosecond Ti-sapphire laser was used to excite four different commonly used fluorochromes, namely Hoechst 33342, Fluo-4, PI and Indo-1, and characteristic fluorescence images were obtained using (455 +/- 15) nm, (540 +/- 15)nm, (580 +/- 16) nm and (500 +/- 15) nm filters respectively. An approach to exciting with a single wavelength, staining with two different fluorochromes, and collecting fluorescence in two separate channels was employed to study mouse preimplantation embryos by 3D and 4D real-time imaging. Combined with the merits of two-photon excitation fluorescence imaging, such as better penetration, less photon-damage, and higher signal-to-noise ratio, this approach was supposed to be a novel multi-parameter investigating tool for mouse preimplantation embryos development study.

Animals↗

PubMed2004

Nociceptive responses of anterior cingulate cortical ensembles in behaving rats.

OBJECTIVE: To confirm the role of anterior cingulated cortices (ACC) in the coding of pain affect by exploring the neural ensemble coding pattern within the anterior cingulate cortex in behaving rats with a multichannel recording technique. METHODS: In five adult male Sprague-Dawley rats, two arrays of eight stainless steel microwires were bilaterally implanted into ACC. Noxious radiant heat stimulation was applied to the tail, bilateral fore-paws and hind-paws of freely moving rats. Neuroelectric signals were obtained from the microwires and sent to a multichannel recording device via cables and connectors. The time stamps of neuronal activities were stored on a personal computer for off-line analysis. RESULTS: Noxious heat stimuli evoked predominantly excitatory and sustained neural activity within ACC, reflecting the processing of pain unpleasantness; pain-related anticipatory responses could be seen near the stimulation start, indicating the behavioral preparation for escape; ACC neurons had broad receptive fields by showing quite similar pain-related responses to stimuli on either side of the hind-paw, suggesting that they are not eligible for the localization of a stimulus. CONCLUSION: ACC has played a major role in processing the affective-motivational aspect of pain.

Animals↗

PubMed2005

Surface properties and cell response of low metal ion release Ti-6Al-7Nb alloy after multi-step chemical and thermal treatments.

Ti-6Al-7Nb samples treated by innovative multi-step chemical and thermal processes were characterized in order to evaluate their surface properties and cell interaction. The main object was to asses if the treatments were effective in order to obtain a surface presenting at the same time bone-like apatite induction ability, low metal ion release, good cell response and high protein binding. The morphology, crystallographic structure, porosity and wettability of the treated materials were investigated, as well as their interaction with simulated body fluid during soaking for different times. Cytotoxicity, protein adsorption tests and in vitro fibroblast and osteoblast-like cell cultures were also performed.

Animals↗

PubMed2002

Surface modification of titanium carbide with carbyne-containing nanocoatings.

The aim of this research is to investigate a novel approach to surface engineering of biomaterials that are based on transition metals of the groups IVA-VIA. The approach taken relies on the fact that, during the electropolishing of TiC surfaces, the removal of Ti atoms from the TiC surface surpasses that of C atoms. This leads to enrichment of the TiC surface with carbon. Transmission electron microscopic investigation showed that carbon-based films contain carbynes in the form of nanorod-like clusters with lengths in the range of 5-100 nm. This carbyne-containing layer is 50-100 nm thick. It was generalized that carbyne-containing nanofilms are formed on the carbide surface of transition metals of groups IVA-VIA during electropolishing. Since carbynes, being one-dimensional chain-like structures [(-C identical to C-)n/(=C=C=)n] with sp1 carbon-carbon hybridization, have the highest degree of biocompatibility because of their biological activity, the development of such surface bioengineering with carbynes extends applications of biomaterials based on transition metals of the groups IVA-VIA.

Carbamates↗

PubMed1998

Effect of upper airway negative pressure on inspiratory drive during sleep.

To determine the effect of upper airway (UA) negative pressure and collapse during inspiration on regulation of breathing, we studied four unanesthetized female dogs during wakefulness and sleep while they breathed via a fenestrated tracheostomy tube, which was sealed around the permanent tracheal stoma. The snout was sealed with an airtight mask, thereby isolating the UA when the fenestration (Fen) was closed and exposing the UA to intrathoracic pressure changes, but not to flow changes, when Fen was open. During tracheal occlusion with Fen closed, inspiratory time (TI) increased during wakefulness, non-rapid-eye-movement (NREM) sleep and rapid-eye-movement (REM) sleep (155 +/- 8, 164 +/- 11, and 161 +/- 32%, respectively), reflecting the removal of inhibitory lung inflation reflexes. During tracheal occlusion with Fen open (vs. Fen closed): 1) the UA remained patent; 2) TI further increased during wakefulness and NREM (215 +/- 52 and 197 +/- 28%, respectively) but nonsignificantly during REM sleep (196 +/- 42%); 3) mean rate of rise of diaphragm EMG (EMGdi/TI) and rate of fall of tracheal pressure (Ptr/TI) were decreased, reflecting an additional inhibitory input from UA receptors; and 4) both EMGdi/TI and Ptr/TI were decreased proportionately more as inspiration proceeded, suggesting greater reflex inhibition later in the effort. Similar inhibitory effects of exposing the UA to negative pressure (via an open tracheal Fen) were seen when an inspiratory resistive load was applied over several breaths during wakefulness and sleep. These inhibitory effects persisted even in the face of rising chemical stimuli. This inhibition of inspiratory motor output is alinear within an inspiration and reflects the activation of UA pressure-sensitive receptors by UA distortion, with greater distortion possibly occurring later in the effort.

Air Pressure↗

PubMed1998

Titanium surface roughness alters responsiveness of MG63 osteoblast-like cells to 1 alpha,25-(OH)2D3.

Surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells. This study examined whether surface roughness alters cellular response to circulating hormones such as 1 alpha,25-(OH)2D3. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then were machined and acid-etched (MA). Ti disks also were sandblasted (SB), sandblasted and acid etched (CA), or plasma sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were: PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on standard tissue culture polystyrene (plastic) or the Ti surfaces and then treated for 24 h with either 10(-8) M or 10(-7) M 1 alpha,25-(OH)2D3 or vehicle (control). Cellular response was measured by assaying cell number, cell layer alkaline phosphatase specific-activity, and the production of osteocalcin, latent (L) TGF beta, and PGE2. Alkaline phosphatase activity was affected by surface roughness; as the surface became rougher, the cells showed a significant increase in alkaline phosphatase activity. Addition of 1 alpha,25-(OH)2D3 to the cultures caused a dose-dependent stimulation of alkaline phosphatase activity that was synergistic with the effect caused by surface roughness alone. 1 alpha,25-(OH)2D3 also caused a synergistic increase in osteocalcin production as well as local factor (LTGF beta and PGE2) production on the rougher CA, SB, and PS surfaces, but it had no effect on the production on smooth surfaces. The inhibitory effect of surface roughness on cell number was not affected by 1 alpha,25-(OH)2D3 except on the SB surface. 1 alpha,25-(OH)2D3 decreased cell number, increased alkaline phosphatase activity and osteocalcin production, and had no effect on LTGF beta or PGE2 production by MG63 cells grown on tissue culture polystyrene. These data suggest that bone cell response to systemic hormones is modified by surface roughness and that surface roughness increases the responsiveness of MG63 cells to 1 alpha,25-(OH)2D3. They also suggest that the endocrine system is actively involved in normal bone healing around implants.

Alkaline Phosphatase↗

PubMed2004

Determination of amphetamine in rat brain by in vivo microdialysis and ion-pairing liquid chromatography with electrospray tandem mass spectrometry.

An ion-pairing liquid chromatography/electrospray tandem mass spectrometry (LC/ES-MS/MS) method with in vivo microdialysis for the determination of amphetamine in rat brain has been developed. A microdialysis probe was surgically implanted into the striatum of the rat and artificial cerebrospinal fluid (aCSF) was used as the perfusion medium. Samples were collected and then analyzed off-line by LC/ES-MS/MS. A reversed-phase C18 column was employed for LC separation and MS/MS was utilized for detection. Trifluoroacetic acid (TFA) was added to the mobile phase (acetonitrile/water) as an ion-pairing reagent. Detection was by ES-MS/MS directly, and no post-column addition of organic modifier was needed. Dual linear ranges were determined from 0.1-0.5 microg/mL and 0.005-0.1 microg/mL, respectively. The detection limit, based on a signal-to-noise ratio of 3, was 0.001 microg/mL (5 nM). Good precision and accuracy were obtained. The applicability of this newly developed method was demonstrated by continuous monitoring of amphetamine concentrations in rat brain. Amphetamine reached a maximum concentration of 0.086 +/- 0.017 microg/mL over 20-40 min after a single 3.0 mg/kg intraperitoneal administration.

Amphetamine↗

PubMed2002

The electrochemical behavior of Ti-6Al-7Nb alloy with and without plasma-sprayed hydroxyapatite coating in Hank's solution.

The electrochemical behavior of Ti-6Al-7Nb alloy with and without plasma-sprayed hydroxyapatite (HA) coating was investigated in Hank's balanced salt solution at 37 degrees C. This behavior was evaluated by analyzing both corrosion potential variation with time curves and potentiodynamic polarization curves. The polarization curves were determined both in aerated and deaerated solutions. It was found that the corrosion potential of uncoated sample is always more noble than that of coated one. In aerated solutions the polarization curve of a coating-free alloy presents a typical passive/transpassive behavior, which is absent in the HA-coated alloy and in deaerated solution for both coated and uncoated conditions. In the latter polarization curves, the current density continuously increases with the increase of the potential. The corrosion rate determined from polarization curves is higher for HA-coated alloys, but it still can be considered negligible (less than 1 microA/cm(2)).

Alloys↗