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

PLoS One. 2020 Apr 7;15(4):e0230493. doi: 10.1371/journal.pone.0230493. eCollection 2020.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcinosis / blood
  • Calcinosis / diagnosis
  • Calcium Phosphates / blood*
  • Humans
  • Kidney Failure, Chronic / pathology
  • Lab-On-A-Chip Devices
  • Microfluidics / instrumentation
  • Microfluidics / methods*
  • Polymers / chemistry*
  • Renal Dialysis
  • Signal-To-Noise Ratio
  • Temperature

Substances

  • Calcium Phosphates
  • Polymers
  • calcium phosphate

Grants and funding

This work was supported by a fund of the Federal Ministry for economic affairs and energy of the Federal Republic of Germany (https://www.bmwi.de/Redaktion/EN/Artikel/SME-Sector/technology-neutral-project-support-01.html; grant number ZIM ZF4429601AJ7) awarded to WKS and by grants awarded to WJD by the IZKF Aachen of the Medical Faculty of RWTH Aachen (https://www.izkf.rwth-aachen.de/index.php/en/izkf-aachen) and by the German Research Foundation (https://www.dfg.de/en/index.jsp; DFG SFB/TRR219-Project C-03). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. AP and WJD are co-founders and share holders of Calciscon AG, a start-up company based at Nidau, Switzerland. AP is an employee of Calciscon AG, who provided support in the form of salaries for author AP, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.