Rapid calcification propensity testing in blood using a temperature controlled microfluidic polymer chip.

Phosphate toxicity is a major threat to cardiovascular health in chronic kidney disease. It is associated with oxidative stress, inflammation and the accumulation of calcium phosphate commonly known as calcification in soft tissues leading to functional disorders of blood vessels. An improved calcification propensity test for the assessment of phosphate toxicity was developed, which measures the velocity of calcium phosphate mineralization from colloidal precursors in vitro. This so called T50 test measures the transformation from a primary into a secondary form of nanosized colloidal plasma protein-calcium phosphate particles known as calciprotein particles. The T50 test in its previous form required a temperature controlled nephelometer and several hours of continuous measurement, which precluded rapid bed side testing. We miniaturized the test using microfluidic polymer chips produced by ultrasonic hot embossing. A cartridge holder contained a laser diode for illumination, light dependent resistor for detection and a Peltier element for thermo control. Increasing the assay temperature from 37°C to 75°C reduced the T50 test time 36-fold from 381 ± 10 min at 37°C to 10.5 ± 0.3 min at 75°C. Incorporating sputtered micro mirrors into the chip design increased the effective light path length, and improved signal-to-noise ratio 9-fold. The speed and reproducibility of the T50 chip-based assay run at 75°C suggest that it may be suitable for rapid measurements, preferably in-line in a dialyser or in a portable microfluidic analytic device with the chip inserted as a disposable cartridge.

Emerging drugs for hyperphosphatemia.

Cardiovascular mortality is the leading cause of death in the uremic patient. Hyperphosphatemia is considered an independent risk factor associated with cardiovascular morbidity and mortality in dialysis patients. As phosphate control is not efficient with diet or dialysis, phosphate binders are commonly prescribed in patients with chronic renal failure. Aluminum salts, the first phosphate binders, even if effective, have several side effects due to their deposition in CNS, bone and hematopoietic cells. Calcium-containing phosphate binders, used in the last 15 years, increase total body calcium load and may exacerbate metastatic calcification, thus, increasing the risk of cardiovascular mortality. Recently two new compounds non-aluminum and non-calcium phosphate binders, sevelamer hydrochloride and lanthanum carbonate, have been introduced. Sevelamer, besides the effect on phosphate, has been associated with reduction of coronary and aortic calcification and with other pleiotropic effects especially on lipid metabolism. Lanthanum carbonate has similar phosphate control to calcium-based binders with less incidence of hypercalcemia but long-term clinical studies are needed for testing long-term exposure. Recently the authors found in dialysis patients, that salivary phosphorus correlated with serum phosphorus. Therefore, they supposed that the use of salivary phosphate binders could reduce its absorption and represent a chance for reducing the serum phosphate concentration in uremic patients.

Behavior in simulated body fluid of calcium phosphate coatings obtained by laser ablation.

Three types of calcium phosphate coatings onto titanium alloy substrates, deposited by the laser ablation technique, were immersed in a simulated body fluid in order to determine their behavior in conditions similar to the human blood plasma. Neither the hydroxyapatite coating nor the amorphous calcium phosphate coating do dissolve and the alpha-tricalcium phosphate phase of the coating of beta-tricalcium phosphate with minor alpha phase slightly dissolves. Precipitation of an apatitic phase is favored onto the hydroxyapatite coating and onto the coating of beta-tricalcium phosphate with minor alpha phase. Onto the titanium alloy substrate reference there is also precipitation but at larger induction times. However, onto the amorphous calcium phosphate coating no precipitate is formed.

Non-decay type fast-setting calcium phosphate cement: setting behaviour in calf serum and its tissue response.

Non-decay type fast-setting calcium phosphate cement (nd-FSCPC) was evaluated in terms of its setting behaviour in calf serum and its tissue response to investigate the feasibility of its clinical use in surgical applications. Non-decay type cements were prepared by adding various amounts of sodium alginate to the liquid phase of base cements, fast-setting calcium phosphate cement (FSCPC) and conventional calcium phosphate cement (c-CPC). Cement pastes were immersed in serum at 37 degrees C immediately after mixing, and decay behaviour, setting time and mechanical strength were measured to evaluate the possibility of their use in surgical applications. Also, nd-FSCPC was implanted into rat subcutaneous tissue for the initial evaluation of biocompatibility of this potential bioactive cement. nd-FSCPC set in approximately 6-7 min in serum, even when the cement paste was immersed in the serum immediately after mixing, whereas c-CPC and FSCPC decayed completely upon immersion. nd-FSCPC transforms to hydroxyapatite (HA) within 24 h and shows a diametral tensile strength of approximately 4-5 MPa. As a result of transformation to HA, nd-FSCPC showed excellent tissue response when implanted subcutaneously in rats. We conclude that nd-FSCPC has good potential value for use in orthopaedics, plastic and reconstructive surgery, and oral and maxillofacial surgery, where the cement is exposed to blood.

Effect of subvoxel processes on non-destructive characterization of ß-tricalcium phosphate bone graft substitutes.

The geometric features of bone graft substitutes, such as the pore and pore interconnection sizes, are of paramount importance for their biological performance. Such features are generally characterized by micro-computed tomography (µCT). Unfortunately, the resolution of µCT is often too limited. The aim of this study was to look at the effect of µCT resolution on the geometric characterization of four different bone graft substitutes. An attempt was also made to improve the characterization of these materials by applying a subvoxelization algorithm. The results revealed that both approaches increased the accuracy of the geometric characterization. They also showed that the interconnection size in particular was affected. Comparing the results obtained from the scanned and numerical subvoxelization datasets revealed a minor difference of less than 2.5% for the porosity values. The difference for the pore sizes was up to 10%. Considerable differences of up to 35-50% were found for the interconnection sizes. The present study demonstrates how complex geometric characterization is and how important it is for biomaterial researchers to be aware of the impact of µCT resolution on the pore and pore interconnection sizes.