Application of X-ray microanalysis to the study of cell physiology in cells attached to biomaterials

Roomans, G. M.

doi:10.22203/ecm.v003a01

Cited by 12 publications

(18 citation statements)

References 34 publications

(32 reference statements)

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“…Moreover, high concentrations of sulfur and phosphorous and moderate concentrations of magnesium and calcium were also found at P6. From a microanalytical point of view, phosphorus is an element that is associated with cell mass and organic constituents, nucleic acids contents and the level of cellular phosphorylation, and cells characterized by a severe structural damage show a decrease in intracellular concentrations of phosphorus [21]. Therefore, the high values found at P6 would confirm the vital status of these cells, whereas the decrease at P7 could be related to a possible structural damage of these cells.…”

Section: Discussionmentioning

confidence: 99%

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

et al. 2012

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Temporo-mandibular joint disc disorders are highly prevalent in adult populations. Autologous chondrocyte implantation is a well-established method for the treatment of several chondral defects. However, very few studies have been carried out using human fibrous chondrocytes from the temporo-mandibular joint (TMJ). One of the main drawbacks associated to chondrocyte cell culture is the possibility that chondrocyte cells kept in culture tend to de-differentiate and to lose cell viability under in in-vitro conditions. In this work, we have isolated human temporo-mandibular joint fibrochondrocytes (TMJF) from human disc and we have used a highly-sensitive technique to determine cell viability, cell proliferation and gene expression of nine consecutive cell passages to determine the most appropriate cell passage for use in tissue engineering and future clinical use. Our results revealed that the most potentially viable and functional cell passages were P5–P6, in which an adequate equilibrium between cell viability and the capability to synthesize all major extracellular matrix components exists. The combined action of pro-apoptotic (TRAF5, PHLDA1) and anti-apoptotic genes (SON, HTT, FAIM2) may explain the differential cell viability levels that we found in this study. These results suggest that TMJF should be used at P5–P6 for cell therapy protocols.

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Section: Discussionmentioning

confidence: 99%

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

et al. 2012

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“…We also expressed our data for Na, Cl, and K composition as a ratio of P concentration to obviate possible technical artifacts due to overpenetration of the electron probe in the sample, which may occur when EPXMA is used to analyze cells growing on solid supports (Roomans, 2002). According to criteria established by Abraham et al (1985), the use of elemental P ratios was possible because no significant differences in intracellular [P] between untreated control and STS‐treated cells.…”

Section: Discussionmentioning

confidence: 99%

Changes in intracellular sodium, chlorine, and potassium concentrations in staurosporine‐induced apoptosis

Arrebola

Zabiti

Cañizares

et al. 2005

Journal Cellular Physiology

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Ion gradients across the plasma membrane, fundamentally K(+), play a pivotal role in the execution phase of apoptosis. However, little is known about other monovalent anions (Cl(-)) or cations (Na(+)) in apoptosis. In addition, the relationship between changes in total ion composition and morphological and biochemical events are poorly understood. We investigated simultaneous changes in sodium (Na), chlorine (Cl), and potassium (K) concentrations in stauroporine-induced apoptosis by quantitative electron probe X-ray microanalysis (EPXMA) in single cells. Apoptotic cells identified unequivocally from the presence of chromatin condensation in backscattered electron images were characterized by an increase in intracellular Na, a decrease in intracellular Cl and K concentrations, and a decrease in K/Na ratio. The ouabain-sensitive Rb-uptake assay demonstrated a net decrease in Na(+)/K(+)-ATPase activity, suggesting that increases in Na and decreases in K and the K/Na ratio in apoptotic cells were related with inhibition of the Na(+)/K(+)-ATPase pump. These changes in diffusible elements were associated with externalization of phosphatidyl serine and oligonucleosomal fragmentation of DNA. This alteration in ion homeostasis and morphological hallmarks of apoptosis occur in cells that have lost their inner mitochondrial transmembrane potential and before the plasma membrane becomes permeable.

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“…In parallel with the decrease of the total K + and Na + content, a pronounced reduction in the Cl and P content was also found in apoptotic human monocytes [54]. As apoptosis and necrosis are both associated with a decrease in the K + /Na + ratio, the differences of element ratios may not allow the discrimination between those two types of cell death [52]. According to our observations, the sum of K + and Na + contents is lower in shrunken apoptotic cells vs normal cells.…”

Section: Discussionmentioning

confidence: 99%

Thymocyte K<sup>+</sup>, Na<sup>+</sup> and Water Balance During Dexamethasone- and Etoposide-Induced Apoptosis

Yurinskaya¹,

Moshkov²,

Rozanov³

et al. 2005

Cell Physiol Biochem

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The mechanism of apoptotic cell volume decrease was studied in rat thymocytes treated with dexamethasone (Dex) or etoposide (Eto). Cell shrinkage, i.e. dehydration, was quantified by using buoyant density of the thymocytes in a continuous Percoll gradient. The K⁺ and Na⁺ content of cells from different density fractions were assayed by flame emission analysis. Apoptosis was tested by microscopy and flow cytometry of acridine orange stained cells as well as by flow DNA cytometry. Treatment of the thymocytes with 1 µM Dex for 4-5.5 h or 50 µM Eto for 5 h resulted in the appearance of a new distinct high-density cell subpopulation. The cells from this heavy subpopulation but not those with normal buoyant density had typical features of apoptosis. Apoptotic increase of cell density was accompanied by a decrease in cellular K⁺ content, which exceeded the simultaneous increase in cellular Na⁺ content. Cellular loss of K⁺ contributed to most of the estimated loss of cellular osmolytes, but owing to the parallel loss of cell water, the decrease in cytosolic K⁺ concentration was less than one third. Due to gain of Na⁺ and loss of cell water the cytosolic Na⁺ concentration in thymocytes rose following treatment with Dex (5.5 h) or Eto (5 h) by a factor of about 3.6 and 3.1, respectively.

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Application of X-ray microanalysis to the study of cell physiology in cells attached to biomaterials

Cited by 12 publications

References 34 publications

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

A Combined Approach for the Assessment of Cell Viability and Cell Functionality of Human Fibrochondrocytes for Use in Tissue Engineering

Changes in intracellular sodium, chlorine, and potassium concentrations in staurosporine‐induced apoptosis

Thymocyte K<sup>+</sup>, Na<sup>+</sup> and Water Balance During Dexamethasone- and Etoposide-Induced Apoptosis

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