In‐vitro analysis of early calcification in aortic valvular interstitial cells using Laser‐Induced Breakdown Spectroscopy (LIBS)

Abstract

Keywords: calcific aortic valve disease, valvular interstitial cells, calcium deposition, laser induced breakdown spectroscopy (LIBS)Calcific aortic valve disease (CAVD) is a major cardiovascular disorder caused by osteogenic differentiation of valvular interstitial cells (VICs) within aortic valves. Conventional methods like colorimetric assays and histology fail to detect small calcium depositions during in-vitro VIC cultures. Laser-induced breakdown spectroscopy (LIBS) is a robust analytical tool used for inorganic materials characterizations, but relatively new to biomedical applications. We employ LIBS, for the first time, for quantitative in-vitro detection of calcium depositions in VICs at various osteogenic differentiation stages. VICs isolated from porcine aortic valves were cultured in osteogenic media over various days. Colorimetric calcium assays based on arsenazo dye and Von Kossa staining measured the calcium depositions within VICs. Simultaneously, LIBS signatures for Ca I (422.67 nm) atomic emission lines were collected for estimating calcium depositions in lyophilized VIC samples. Our results indicate excellent linear correlation between the calcium assay and our LIBS measurements. Furthermore, unlike the assay results, the LIBS results could resolve calcium signals from cell samples with as early as 2 days of osteogenic culture. Quantitatively, the LIBS measurements establish the limit of detection for calcium content in VICs to be~0.17 AE 0.04 mg which indicates a 5-fold improvement over calcium assay. Picture: Quantitative LIBS enables in-vitro analysis for early stage detection of calcium deposition within aortic valvular interstitial cells (VICs).

Jolly green MOF: confinement and photoactivation of photosystem I in a metal–organic framework

“…The LIBS experimental set-up is described in detail elsewhere. 53,56,57 Briey, it uses a Q-switched Nd-YAG laser of a nominal wavelength of 1064 nm operating at 200 mJ per pulse, and a pulse width of 8 ns (Make: Insight Model: 122551-R) to thermally ablate the samples to collect atomic emission signatures of the constituent analyte species. To improve the statistical average of the signal intensity for each analyte of interest, spectral data over $40 shots were collected.…”

Hybrid nanocomposites of nanostructured Co3O4interfaced with reduced/nitrogen-doped graphene oxides for selective improvements in electrocatalytic and/or supercapacitive properties

“…The Co concentrations in the composites under study were measured via inductively coupled plasma optical emission spectroscopy (ICP-OES) (Make: Perkin Elmer; Model: Optima 4300 DV) and our in-house developed quantitative laser-induced breakdown spectroscopy (Q-LIBS) as described elsewhere. [72][73][74][75] X-ray diffraction (XRD) was carried out on a Phillips X'Pert-Pro diffractometer equipped with a Cu Ka source at 45 kV and 40 mA. The Fourier transformed infrared (FTIR) spectrum was obtained using a Nicolet 6700 FT-IR Spectrometer at the wavenumber range of 400 to 3000 cm À1 .…”

Quantification of toxic metallic elements using machine learning techniques and spark emission spectroscopy

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