Simulated Physiological Solution Research Articles

The effect of calcium phosphate ceramic composition and structure on in vitro behavior. II. Precipitation

The formation of a biologically equivalent carbonate-containing apatite on the surface of synthetic calcium phosphate ceramics (CPC) may be an important step leading to bonding with bone. Reactions of several single phases CPCs upon immersion into a simulated physiologic solution (SPS) with an electrolyte composition of human plasma were determined. The CPCs covered a wide range of solution stabilities from low-soluble hydroxyapatites (HA) to metastable tricalcium phosphates (TCP) and tetracalcium phosphate (TTCP). Changes in chemical compositions of SPS and infrared spectral features after CPC immersion were analyzed. New phase formation was observed on all the CPCs. However, kinetics, compositions, and structures of the new phases were significantly different. The studied CPCs can be characterized by the time to new phase formation in vitro; the minimum time for measurable precipitate formation was found to increase in the order: not-well-crystallized HAs < well-crystallized HAs < alpha-TCP, TTCP < beta-TCP. Among the CPCs only not-well-crystallized HAs led to immediate new phase formation. The metastable CPCs, beta-TCP, alpha-TCP, and TTCP required an induction time during which dissolution occurred. beta-TCP showed the longest induction time and the lowest lattice ion uptake rate of all the CPCs tested. Only the not-well-crystallized HAs elicited immediate formation of carbonated HA. The well-crystallized HAs and beta-TCP did not elicit carbonated apatite formation within the time frame of the experiment. Instead, intermediate phases were formed. On alpha-TCP amorphous calcium phosphate (ACP) with a relatively low carbonate content was formed. TTCP was found to transform extensively to poorly crystallized carbonated apatite after 2 days of immersion.

The effect of dissolution on plasma sprayed hydroxylapatite coatings on titanium

Plasma sprayed hydroxylapatite (HA) coated titanium specimens were immersed into Ca-free Hank's balanced salt solution for periods of l, 2, 4, and 6 weeks. At each of the respective time intervals the HA coatings were analyzed with X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and Raman spectroscopy to determine the effect of dissolution on the structure and composition of the coatings. At the 2- and 6-week intervals, additional samples were removed from solution and shear tested to evaluate the effect of dissolution on the coating-to-substrate interfacial bond strength. XRD results revealed that the plasma sprayed coatings retained their basic apatitic structure throughout the 6-week period in solution. There was, however, a trend towards a more definitive baseline, suggesting a loss of amorphous material by dissolution. FTIR and Raman analyses of the as-sprayed and dissolution specimens showed that the phosphate groups were not lost during the time in solution; however, there was a decrease in hydroxyl content as a result of dissolution mechanisms. FTIR also revealed an increase in carbonate content within the coating during immersion in the simulated physiological balanced salt solution. The average shear bond strengths for the as-sprayed, 2-, and 6-week dissolution specimens were 14·82 ± 3·52 MPa, 12·51 ± 3·41 MPa, and 12·54 ± 2·02 MPa, respectively. Duncan's multiple range test confirmed that the shear bond strength was not significantly reduced after 6 weeks in solution, but there was evidence that suggested a decrease in interfacial bond strength as a result of dissolution mechanisms.

The effect of residual glassy phase in a bioactive glass-ceramic on the formation of its surface apatite layerin vitro

A bioactive glass containing (in wt%) SiO2 48, P2O5 9.5, Na2O 20 and CaO 22.5 was transformed into a glass-ceramic through a heat treatment. The apatite formation on the surface of this glass-ceramic was examined in a simulated physiological solution. The data from X-ray diffraction, infrared reflection spectroscopy, scanning electron microscopy together with energy-dispersive X-ray analysis and composition imaging of backscattered electrons showed that the formation of the surface apatite layer depends on the relative amount of residual glassy phase in the glass-ceramic. The apatite layer was found to formin vitro on its surface if the glass-ceramic contained a residual glassy phase in a relative proportion more than a limiting volume. It lay on a layer rich in silica. However, only, a silica-rich layer was developed within the surface region of the glass-ceramic during the interaction with solution if the glass was almost completely crystallized. It is proposed that the apatite formation on the surface of the glass-ceramic is mainly caused by its residual glass. The residual glass facilitating apatite formation is considered to provide a negatively charged surface developed during its corrosion in the surrounding solution. The negatively charged surface attracts calcium ions and creates a solution within the glass — solution interface that is highly supersaturated with respect to hydroxyapatite. This leads to the formation of apatite on the surface of the glass-ceramic.

Compositional dependence of calcium phosphate layer formation in fluoride Bioglasses.

Bioglasses form a double layer composed of apatite and a silica-rich layer when placed in a simulated physiological solution as well as in living tissue [A.E. Clark, C.G. Pantano, and L. L. Hench, "Auger spectroscopic analysis of bioglass corrosion films," J. Am. Ceram. Soc., 59(1-2), 37-39 (1976).]. In the present work, the mechanisms of the calcium phosphate layer and the silica-rich layer formation of fluoride Bioglasses in Tris-buffer solution are studied as a function of the SiO2 content. Fourier Transform Infrared Reflection Spectroscopy (FTIRS) is used to investigate the mechanism of formation of calcium phosphate and silica-rich layers on the glass surface. Ion concentration in reacted solution and elemental depth profiles are obtained by Induced Coupled Plasma Atomic Emission Spectrometry (ICP) and Auger Electron Spectroscopy (AES), respectively. Si--O bonds with one nonbridging oxygen and Si--O--Si bonds form at the early stage of reaction. Strong phosphorus ion uptake occurs when an amorphous calcium phosphate layer crystallizes. Glasses with high silica content (conventional glass) form the silica-rich layer first followed by a calcium phosphate layer on top. However, glasses with low silica content (invert glass) form both layers simultaneously. The rate of apatite formation decreases with increasing SiO2 content, especially in the region of conventional glass compositions. Ion release rates decreases as SiO2 content increases, with a significant change occurring at the compositional boundary between invert and conventional glasses.

Compositional variations in the surface and interface of calcium phosphate ceramic coatings on Ti and Ti-6Al-4V due to sintering and immersion

The compositions of the surface and the interface of calcium phosphate ceramic (CPC) coatings electrophoretically deposited and sintered on titanium or its alloy, were determined by scanning Auger electron spectroscopy before and after 4 wk of immersion in a simulated physiological solution. In the CPC coatingmetal interfaces, the phosphorus diffused beyond the titanium oxide layer. The phosphorus concentration in the interface followed a Gaussian distribution for both unalloyed and alloyed titanium. The diffusion depleted P in the ceramic adjacent to the metal. The surface of the ceramic, however, was substantially unchanged. A major change in the compositional depth profiles was induced by immersion: thick and uniform titanium phosphide layers of constant composition were observed on the Ti-based metal substrates.

The electrochemistry of a glass surface and its application to bioactive glass in solution

Hydroxyapatite (HAP) precipitates on the surface of bioactive glasses after their exposure to a simulated physiological solution and the precipitation changes with the ionic species and the pH value in the solution. The formation of the electric double layer around the glass surface can be deduced theoretically from these results.

Preparation of zirconia-toughened bioactive glass-ceramics

Bioactive glass-ceramics toughened by tetragonal zirconia polycrystal (TZP) were prepared by hot-pressing mixed powders of the MgO-CaO-P2O5-SiO2 glass and TZP containing 20 to 80% alumina. The bending strength and the fracture toughness of the composite materials were improved compared with those of the material without TZP. These composites showed high bending strengths (400 to 500MPa) and high fracture toughness (∼ 2.8MPa m1/2). The existence of a crack deflection mechanism was observed by scanning electron microscopy. After soaking in simulated physiological solution at 100 °C, no phase transformation from tetragonal to monoclinic of TZP in the composites and no degradation in bending strength occurred.

Compositional dependence of formation of an apatite layer on glass-ceramics in simulated physiological solution

In simulated physiological solution, an apatite layer is formed on the surface of apatite-containing glass-ceramics having the ability to bond to living bone. In this study, the influence of composition in the system CaO-P2O5-SiO2-MgO, Al2O3 on apatite layer formation is investigated. On CaO-P2O5-SiO2 glass-ceramics, an apatite layer was formed rapidly in simulated physiological solution. However, a solution containing an excess of Mg2+ prevented apatite layer formation. On glass-ceramics containing MgO, the amount of apatite formed on the surface decreased. An apatite layer was not formed on glass-ceramics containing Al2O3. The prevention of apatite layer formation on glass-ceramics containing MgO is attributed to an increase of Mg2+ concentration in the solution. It is thought that glass-ceramics containing Al2O3 form are Al2O3-rich layer, and that this layer prevents the formation of an apatite layer.

Doping influence on the interaction between a bioactive glass and a simulated physiological solution: Chemical and EPR tests

The adhesion between bioactive glass and metal is improved by the addition of small amounts of transition ion oxides. Because it is difficult to carry outin vivo tests to check with continuity, chemical information about the interactions of the different doped glasses with living tissues, the behaviour of the different doped bioactive glasses in a liquid simulating physiological conditions was studied. Chemical and EPR analyses show that the mineralization phenomena around the bioglasses in the tissues also take place at an inorganic level. This simulation shows that the ions released from the different vitreous systems are correlated with the adopted doping. In particular an increase in doping with iron involves a decrease in the release rate of the other ions. Furthermore the doping agents allow a control of the release rate and consequently the obtainment of the best biological adaptability.

Anodic polarization behavior of Ti-Ni and Ti-6A1-4V in simulated physiological solutions.

Anodic polarization measurements made in Hanks' physiological solution at 37 degrees C and a pH of 7.4 show titanium materials to be the most passive of the following metals: titanium, Ti-6A1-4V, Ti-Ni (memory alloy), MP35N (Co-Ni-Cr-Mo), Co-Cr-Mo, 316L stainless steel, and nickel. The influence of the amino acids, cysteine, and tryptophan on the corrosion behavior of Ti-Ni and Ti-6A1-4V was studied. Cysteine caused a lower breakdown potential for Ti-Ni, but it did not affect the breakdown of Ti-6A1-4V, although an increase in current density for Ti-6A1-4V was observed. Tryptophan produced no significant effects.

Compositional dependence of the formation of calcium phosphate films on bioglass.

Bioglass, which has a composition of sodium carbonate, calcium carbonate, phosphorous pentoxide and silica, has been shown to bond to living bone. This ability is dependent on controlled surface reactions. Investigators with 45S5 bioglass have demonstrated that the formation of a SiO2-rich layer and a calcium phosphate film on its surface in an aqueous environment is associated with the film bonding the bioglass to bone. The objects of this research were: 1. To study SiO2 dependence on the formation of a silica-rich layer and calcium phosphate films on a bioglass surface in a simulated physiological solution, and 2. To establish a correlation between in vitro surface reactions and in vivo bonding ability. It was discovered that three types of reactions occur in a simulated physiological solution depending on bioglass composition: 1. A calcium phosphate film and SiO2-rich layer form simultaneously and the reaction rate is fast for bioglasses which have a lower content of SiO2 (approximately 46 mol% SiO2). 2. A SiO2-rich layer forms first and a calcium phosphate film develops later between the aqueous environment and the SiO2-rich layer for bioglasses whose SiO2 content is between 46--55 mol %. 3. A calcium phosphate film does not form for glasses whose SiO2 content is more than 60 mol %.

Degradation resistance of some candidate composite biomaterials.

The degradation resistance of matrix, fiber and composite systems which we have been studying as candidate orthopedic materials has been examined in two appropriate environments. Both resistance to steam sterilization in an autoclave environment and resistance to a simulated physiologic solution have been studied. In the autoclave study, samples were placed in a pressure cooker at 123 degrees C for differing amounts of time and tested for retention of mechanical properties. Results indicate that most of the materials tested could be autoclaved several times, as long as autoclave times did not exceed 1 hr. Longer autoclave times result in an accelerated degradation and loss of strength of all materials except the polypropylene. Polysulfone degrades after even the shortest autoclave duration. Resistance to the simulated physiologic environment was tested by measured retention of mechanical properties after immersion times in pseudo-extracellular fluid (PECF) at 37 degrees C for as long as three years. None of the materials showed any significant changes in properties after immersion in the PECF.

Myoelectrical and mechanical activity of isolated canine stomach perfused in vitro with fluorocarbon. Role of glucose, phosphate, calcium and strontium in the perfusate.

Myoelectrical and mechanical activities were recorded from whole isolated canine stomachs perfused, intravascularly, with fluorocarbon emulsion, oxygenated in vitro. The perfusate was composed of an oxygen carrier, fluorcarbon, emulsified with a surfactant added to a simulated physiological solution. Bipolar electrodes and a strain gauge were used for recordings. Stomachs perfused as above for 5 h displayed normal electrical and mechanical activities. 'Normal' meaning identical with those observed under in vivo conditions or during perfusion with homologous blood. Normal response to electrical stimulation of a vagus nerve branch or to intraarterial injection of methacholine or pentagastrin were also observed. These functions of smooth muscles were not altered when glucose-free or glucose-and phosphate-free perfusate were used. In calcium-free perfusate, electrical and mechanical activities were absent and no responses to stimulants used were recorded. When calcium was added to the perfusate, these functions were restored to normal. Strontium was found to be an adequate substitute for calcium in fluorocarbon perfusate. Gastric secretions were alkaline and contained fluorocarbon emulsion.

Effect of ex vivo perfusion of isolated canine stomach with fluorocarbon on the composition of gastric tissues.

Isolated canine stomachs were perfused for 6 h with fluorocarbon emulsion suspended in a simulated physiological solution of electrolytes with glucose added. Gastric mucosa and extramucosal tissues of perfused stomachs were sampled for biochemical analysis of high energy phosphates, glucose and electrolytes. Comparable samples were also taken from normal canine stomachs, dissected under similar surgical conditions as the stomachs used for perfusion. Gastric tissue ATP, ADP and AMP were reduced but CP was increased in the tissues perfused with fluorocarbon as compared with controls. Gastric tissue water content and sodium were increased but potassium was reduced in a stomach so perfused. The biochemical tests performed were considered as viability tests of organs preserved-perfused in fluorocarbon prior to possible transplantation. Significance of the changes observed is discussed in the light of the current knowledge on energy metabolism.