scholarly journals Application of FTIR and Raman Spectroscopy to Characterisation of Bioactive Materials and Living Cells

2003 ◽  
Vol 17 (2-3) ◽  
pp. 275-288 ◽  
Author(s):  
I. Notingher ◽  
J. R. Jones ◽  
S. Verrier ◽  
I. Bisson ◽  
P. Embanga ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Near Infrared ◽  
Cell Damage ◽  
Living Cells ◽  
Biologically Active ◽  
Bioactive Materials ◽  
45S5 Bioglass ◽  
Ftir And Raman Spectroscopy

Both Fourier Transform Infrared (FTIR) and Raman spectroscopy have been applied to thein vitrocharacterisation of biomaterials, mainly surface reactions leading to the formation of a biologically active hydroxycarbonate apatite (HCA) layer on the sample surface when immersed in simulated body fluids (SBF). The HCA layer indicates the degree of bioactivity of the sample, because it leads to a strong bond between the biomaterial and living tissue. Reflection measurements using FTIR allow quick, non-destructive detection of the HCA layer for solid and powder samples. Due to the low Raman scattering efficiency and low absorption of water in the visible-near infrared region, Raman micro-spectroscopy was successfully used for thein situcharacterisation of 20 and 40µm diameter 45S5 Bioglass®fibres. Thein situcapabilities of the Raman micro-spectrometer have also been extended to the characterisation of living cells attached on bioinert silica and bioactive 45S5 Bioglass®and 58S substrates. Using a high power 785 nm laser, living cells in physiological conditions can be real-time sampled over long periods of time without inducing cell damage and with good signal strength. Cell death can be monitored because it proved to induce strong changes in the Raman signature in the spectral regions 1000–1150 cm–1and 1550–1650 cm–1.

scholarly journals Measurement and analysis of sarcomere length in rat cardiomyocytes in situ and in vitro

2010 ◽  
Vol 298 (5) ◽  
pp. H1616-H1625 ◽  
Author(s):  
G. Bub ◽  
P. Camelliti ◽  
C. Bollensdorff ◽  
D. J. Stuckey ◽  
G. Picton ◽  
...  
Keyword(s):  
Measurement Errors ◽  
Sarcomere Length ◽  
Ex Vivo ◽  
Living Cells ◽  
Perfused Heart ◽  
Experimental Conditions ◽  
Rat Cardiomyocytes ◽  
Two Photon

Sarcomere length (SL) is an important determinant and indicator of cardiac mechanical function; however, techniques for measuring SL in living, intact tissue are limited. Here, we present a technique that uses two-photon microscopy to directly image striations of living cells in cardioplegic conditions, both in situ (Langendorff-perfused rat hearts and ventricular tissue slices, stained with the fluorescent marker di-4-ANEPPS) and in vitro (acutely isolated rat ventricular myocytes). Software was developed to extract SL from two-photon fluorescence image sets while accounting for measurement errors associated with motion artifact in raster-scanned images and uncertainty of the cell angle relative to the imaging plane. Monte-Carlo simulations were used to guide analysis of SL measurements by determining error bounds as a function of measurement path length. The mode of the distribution of SL measurements in resting Langendorff-perfused heart is 1.95 μm ( n = 167 measurements from N = 11 hearts) after correction for tissue orientation, which was significantly greater than that in isolated cells (1.71 μm, n = 346, N = 9 isolations) or ventricular slice preparations (1.79 μm, n = 79, N = 3 hearts) under our experimental conditions. Furthermore, we find that edema in arrested Langendorff-perfused heart is associated with a mean SL increase; this occurs as a function of time ex vivo and correlates with tissue volume changes determined by magnetic resonance imaging. Our results highlight that the proposed method can be used to monitor SL in living cells and that different experimental models from the same species may display significantly different SL values under otherwise comparable conditions, which has implications for experiment design, as well as comparison and interpretation of data.

Potential of solubility, enzymatic methods and NIRS to predict in situ rumen escape protein

1997 ◽  
Vol 45 (2) ◽  
pp. 291-306 ◽  
Author(s):  
J.L. De Boever ◽  
B.G. Cottyn ◽  
J.M. Vanacker ◽  
C.V. Boucque
Keyword(s):  
Phosphate Buffer ◽  
Near Infrared ◽  
Streptomyces Griseus ◽  
Maize Silage ◽  
Near Infrared Reflectance ◽  
In Vitro Method ◽  
Rumen Degradation ◽  
The Difference

The percentage of feed protein escaping rumen degradation was measured by the in situ method (%EPsitu) for 29 compound feeds, untreated and formaldehyde-treated soyabean meal and 12 forages: 3 grass silages, 2 maize silages, fresh grass, grass hay, fodder beets, fresh potatoes, ensiled beet pulp, chopped ear-maize silage and brewers' grains. Loss of particles through bag pores was determined by the difference between the washable fraction (W) and the fraction soluble in borate-phosphate buffer at pH 6.7 (S). W - S was most pronounced for compound feeds (on average 14.4 percentage units), for brewers' grains and maize silages. A correction of %EPsitu, assuming that W - S degrades like the potentially degradable fraction, was not appropriate. Solubility in borate-phosphate buffer after 1 h, enzymic degradability by protease from Streptomyces griseus or ficin after 1, 6 and 24 h and near infrared reflectance spectroscopy (NIRS) (for compound feeds alone) were examined as a routine method to predict %EPsitu. With the buffer and S. griseus the effect of pH (6.7 vs. 8.0) and at pH 8.0 the effect of amount of substrate (500-mg sample vs. 20 mg N) were tested. With ficin, 500-mg samples were incubated at pH 6.7. Predictions were better when compound feeds and forages were considered separately. However, the best in vitro method was different for the 2 feed categories, being solubility in buffer for the compound feeds and enzymic degradation of a constant amount of protein with S. griseus at pH 8.0 for forages. NIRS showed potential to predict %EPsitu of compound feeds, but needs more reference samples. The Dutch feed tables appeared more accurate than the best in vitro method for compound feeds, but was too inaccurate for some forages like fodder beets, maize silage and ear-maize silage.

scholarly journals A near-infrared phosphorescent iridium(iii) complex for imaging of cysteine and homocysteine in living cells and in vivo

RSC Advances ◽  
2017 ◽  
Vol 7 (83) ◽  
pp. 52621-52625 ◽  
Author(s):  
Yongquan Wu ◽  
Renmiao Wu ◽  
Huifang Li ◽  
Hong Zeng ◽  
Yuanyan Li ◽  
...  
Keyword(s):  
Near Infrared ◽  
Living Cells

A novel NIR-emitting iridium(iii) complex was developed to detect Cys/Hcy levels in vitro and in vivo.

scholarly journals Identification of hepatitis C in human blood serum by near-infrared Raman spectroscopy

2008 ◽  
Vol 22 (5) ◽  
pp. 387-395 ◽  
Author(s):  
Jamil Saade ◽  
Marcos Tadeu Tavares Pacheco ◽  
Maira Regina Rodrigues ◽  
Landulfo Silveira Jr.
Keyword(s):  
Raman Spectroscopy ◽  
Hepatitis C ◽  
Blood Serum ◽  
Human Blood ◽  
Near Infrared ◽  
Viral Infections ◽  
Human Blood Serum ◽  
Biochemical Alterations ◽  
Viral Hepatitis C

Hepatitis C has become one of the higher health problems around the world. Near-infrared Raman spectroscopy had been used to spectrally differentiate among health human blood serum from the one with hepatitis C contaminationin vitro. In this study a Raman spectrometer with 80 mW, 830 nm excitation, liquid-nitrogen cooled CCD and imaging spectrograph were used to collect Raman scattering from 24 blood samples (14 healthy and 10 diseased) with collection time of 120 s. It has been used an algorithm based on the Principal Components Analysis (PCA) for main spectral features identification and Mahalanobis distance for blood spectrum classification depending on the serology. It was observed that the highest spectral differences between the two types of human blood serum were found in 1002, 1169, 1262 and 1348 cm−1Raman bands. The spectral analysis using multivariate statistics presented good results when compared to classical diagnosis for viral hepatitis C, showing that Raman spectroscopy can classify human blood serum spectrum in one of the two categories by identifying biochemical alterations that occur in the presence of viral infections.

scholarly journals On the in Vitro Biocompatibility Testing of Bioactive Glasses

Materials ◽  
2020 ◽  
Vol 13 (8) ◽  
pp. 1816 ◽  
Author(s):  
Devis Bellucci ◽  
Elena Veronesi ◽  
Massimo Dominici ◽  
Valeria Cannillo
Keyword(s):  
Predictive Tool ◽  
Bioactive Glasses ◽  
Bioactive Materials ◽  
In Vitro Biocompatibility ◽  
Marrow Mesenchymal Stem Cells ◽  
Biocompatibility Testing ◽  
45S5 Bioglass ◽  
First Time

In this work, a new 3D cellular model—based on human bone marrow mesenchymal stem cells (BM–MSCs)—was used for the first time to test the 45S5 Bioglass® (45S5). Such a model, previously used to evaluate the biologic performance of two novel bioactive glasses, suggested out the regenerative potential of such materials. In fact, BM–MSCs were able both to adhere and colonize the biomaterials, as well as differentiate toward osteoblasts—even in absence of specific growth factors. Surprisingly, BM–MSCs were not able to colonize 45S5 granules (almost no adhesion and/or colonization was observed), and thus, were not capable of providing any osteogenic differentiation. Therefore, the model seems to suggest that the two novel bioactive glasses have a better biologic performance than 45S5. If this hypothesis is confirmed also by in vivo tests, the 3D model may become a predictive tool for discriminating between different potential bioactive materials by comparatively evaluating them, and preliminarily selecting the best ones in relation to their biocompatibility potential—before proceeding with further experiments in vivo. This approach could favor the reduction of costs and time of pre-clinical and clinical trials.

A novel near-infrared xanthene-based fluorescent probe for detection of thiophenol in vitro and in vivo

2020 ◽  
Vol 44 (40) ◽  
pp. 17360-17367
Author(s):  
Yongquan Wu ◽  
Aiping Shi ◽  
Huiying Liu ◽  
Yuanyan Li ◽  
Weican Lun ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Near Infrared ◽  
Living Cells

A novel near-infrared xanthene-based fluorescent probe for detection of thiophenol in living cells and mice.

In vitro quantitation of human femoral artery atherosclerosis using near-infrared Raman spectroscopy

2012 ◽  
Author(s):  
Ava C. Dykes ◽  
Pavlos Anastasiadis ◽  
John S. Allen ◽  
Shiv K. Sharma
Keyword(s):  
Raman Spectroscopy ◽  
Femoral Artery ◽  
Near Infrared

Evaluation of an In Vitro Bioassay for the Detection of Purified Ricin and Castor Bean in Beverages and Liquid Food Matrices

2007 ◽  
Vol 70 (10) ◽  
pp. 2377-2382 ◽  
Author(s):  
JENNIFER L. BRZEZINSKI ◽  
DAVID L. CRAFT
Keyword(s):  
Castor Bean ◽  
Colorimetric Assay ◽  
Cell Damage ◽  
Jurkat Cells ◽  
Biologically Active ◽  
Detection Methods ◽  
Mouse Bioassay ◽  
Animal Testing ◽  
Vitro Assay

The potential use of ricin as a biological weapon in food highlights the necessity for the development of food-specific detection methods. Current methods for the detection of ricin consist of various immunoassays, which detect only one subunit of the ricin toxin and therefore may not be indicative of a biologically active molecule. An in vivo assay, such as a mouse bioassay, can indicate the biological activity of the toxin; however, this method is not feasible for laboratories that do not have animal testing facilities. The purpose of this study was to develop an in vitro assay for the detection of biologically active ricin in beverages and liquid foods. Acidic and high-protein beverages were spiked with either purified ricin or ground castor beans and added to cultured human Jurkat cells. After an overnight incubation, the supernatant was tested for lactate dehydrogenase (LDH) activity with a colorimetric assay. LDH was released from the cytosol upon cell damage and was positively correlated with cell death. Ricin was detectable in all the matrices tested, with a sensitivity of 10 to 100 pg/ml. Biologically active ricin was detectable in all the matrices incubated with ground castor bean material. This method provides a confirmatory way to detect biologically active ricin that can be utilized by laboratories lacking animal facilities.

Inhibition of the alpha-amidation of gastrin: effects on gastric acid secretion

1990 ◽  
Vol 258 (5) ◽  
pp. G810-G814 ◽  
Author(s):  
C. J. Dickinson ◽  
L. Marino ◽  
T. Yamada
Keyword(s):  
Acid Secretion ◽  
Gastric Acid Secretion ◽  
Gastric Acid ◽  
Enzyme Synthesis ◽  
Normal Serum ◽  
Biologically Active ◽  
Sprague Dawley

Formation of biologically active amidated gastrin from glycine-extended progastrin processing intermediates (G-Gly) is achieved via the action of peptidyl-glycyl alpha-amidating monooxygenase. Since this enzyme requires copper for optimal activity, we examined the effects of a known copper chelator, diethyldithiocarbamate (DDC), on gastrin posttranslational processing and gastric acid secretion in vivo. DDC (400 mg.kg-1.day-1 ip X 3 days) administered to male Sprague-Dawley rats decreased antral amidated gastrin content, but increased antral G-Gly content. The ratio of amidated gastrin to G-Gly, which reflects in situ amidating activity, was decreased in DDC-treated rats. In contrast, tissue amidating potential, assayed directly under optimal copper concentrations in vitro, was increased in the antrum and unchanged in the pituitary. DDC markedly increased both basal and gastrin-stimulated gastric acid outputs despite the presence of normal serum amidated gastrin levels. These results suggest that copper chelation with DDC inhibits amidating activity in situ but selectively increases antral amidating enzyme synthesis. The marked increase in acid secretion despite normal circulating amidated gastrin concentrations, combined with the enhanced secretory response to exogenously administered gastrin, suggests the possibility that gastrin receptors are upregulated by the events precipitated via DDC administration.

Sign in / Sign up

Export Citation Format

Share Document