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Spiral and square microstructured surfaces: the effect of the decreasing size of photo-immobilised hyaluronan domains on cell growth
No full textAbstract
Spiral and squared micropatterned surfaces of decreasing dimensions were realized by photo-immobilizing a photoreactive hyaluronan (Hyal) derivative on silanized glass substrates. The microstructured surfaces were observed by atomic force microscope and scanning electron microscope. Scanning electron microscope analysis revealed the presence of a spiral (ranging from 100 microm down to 1 microm in the central part) and a square pattern consisting of a central square of 100 microm x 100 microm and squares of different dimensions decreasing from the centre to the edges of the micropatterned area (2 microm x 1 microm). Three cell types were tested on all the microstructured surfaces: human coronary artery endothelial cells (HCAEC), human dermal fibroblasts (C54), and NIH 3T3 fibroblasts. Cell adhesion analysis demonstrated that HCAEC and C54 did not adhere to the immobilized Hyal on silanized glass but adapted their shape to the different sizes of the square and spiral patterns. Also, in both geometric patterns, the reduction of the adhesive glass width induced C54 to create bonds amongst themselves. NIH 3T3 cells adhered inside the squares and the spiral but reducing the adhesive glass width induced NIH 3T3 to adhere to immobilized Hyal. This fact is explained by the interactions between the cells and the immobilized Hyal as a consequence of the CD44/Hyal binding
Chemistry and topographic domains: micropatterned surfaces. Production, physical-chemical and biological characterisation
No full textThe presence of micro and nano-domains on a surface
allows the manipulation of two fundamental external signals: cellsubstrate
and cell-cell interaction, in order to create a pattern of
highly oriented cells capable of arranging themselves in tissue. Cell
guidance also depends on the different chemical and/or
topographic domains present on a surface and on their geometry.
Thus, structures are realised so that they also contain different
chemical and geometrical domains. Among the many known
methods for modifying surfaces, the photoimmobilisation process
is a useful technique for creating microstructures with both
chemical and topographic patterns.
In most approaches, cells are localized to adhesive regions on a
substrate, thus limiting their use to one cell type. More recently,
approaches have been developed for patterning two or more cell
types in spatially defined co-cultures. These approaches can be used to study the effects of cell shape, cell-matrix interactions, and heterotypic cell-cell
contact on various cell functions. Many studies on patterned co-cultures have involved
the selective adhesion of one cell type compared to the normal adhesion of the other.
In this chapter, the results of our research on micro-topography and cell
behaviour are reviewed and discussed.
Patterns with different geometries have been obtained by photoimmobilisation of
the polysaccharide hyaluronic acid (Hyal) on glass surfaces. The resulting surface
patterns have been utilised to study the influence of microstructures as a function of
chemical, topographic and dimensional properties on both primary and tumoral cell
lines. Cell adhesion and distribution have been analysed as a function of the shape and
area of the microdomains.
Studies carried out with the aim to analyse heterotypic cell-cell interaction as a
function of geometry and dimension domains are also reported. In particular, the
possibility of obtaining co-cultured microstructured surfaces by seeding fibroblasts on
patterned samples with already adhered endothelial cells has been investigated.
The chapter ends with an outline of the results recently obtained by other authors
studying the combined effect of topographic domains and mechanical/gravitational
stress, a very promising topic in the field of cell microenvironment topography
Heterotypic interaction of fibroblasts and endothelial cells on restricted area
No full textThe polysaccharide hyaluronic acid (Hyal) was photoimmobilized on glass surfaces to obtain a pattern with squares and rectangles of different dimensions and chemistry. The microstructured surfaces were characterized by Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM), Attenuated Total Reflection Infrared Spectroscopy (ATR FT-IR), and Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS). Surface analysis revealed the presence of a pattern consisting of alternating glass and Hyal microstructures whose dimensions decreased from the center to the edge of the sample. The behavior of Human Coronary Artery Endothelial Cells (HCAEC) and human tumoral dermal fibroblasts (C54) was studied on these micropatterned surfaces. Neither HCAEC nor C54 adhered to the immobilized Hyal but both adapted their shape to the different sizes of the glass squares and rectangles. The number of adherent HCAEC and C54 depended on the dimensions of both the glass domains and the nuclei of the cells. Co-cultured C54 on HCAEC patterned surfaces showed a heterotypic cell-cell interaction in the same chemical and topographic domain for the first time. In comparison to other techniques for patterning two different cell types, our approach was non cytotoxic and allowed arbitrary geometric patterns to form on different biocompatible substrata
Heterotypic cell-cell interaction on micropatterned surfaces
No full textPurpose. The aim of this paper was to study the influence of chemical and topographical signals on cell behaviour and to obtain a heterotypic cell-cell interaction on microstructured domains.
Methods. The polysaccharide hyaluronic acid (Hyal) was photoimmobilised on glass surfaces in order to obtain a pattern with squares and rectangles of different dimensions and chemistry. The microstructured surfaces were characterised by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The behaviour of Human Coronary Artery Endothelial Cells (HCAEC) and human tumoral dermal fibroblasts (C54) was investigated on these micropatterned surfaces by adhesion studies. Moreover heterotypic interaction among C54 and HCAEC adhered on patterned surfaces was evaluated by time-lapse video microscopy.
Results. Surface analysis revealed the presence of a pattern consisting of alternating glass and Hyal microstructures whose dimensions decreased from the centre to the edge of the sample. Neither HCAEC nor C54 adhered to the immobilised Hyal but both adapted their shape to the different sizes of the glass squares and rectangles. The number of adherent cells depended on the dimensions of both the glass domains and the nuclei of the cells. Co-cultured C54 on HCAEC patterned surfaces showed a heterotypic cell-cell interaction in the same chemical and topographic domain.
Conclusions. A heterotypic cell-cell interaction occurred in the same chemical and topographic micro-domains but in narrow areas only. Moreover, the number of cells adhering to the glass domains and cell morphology depended on the dimensions of both adhesive areas and cell nuclei
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Gonadal ERα/β, AR and TRPV1 gene expression: modulation by pain and morphine treatment in male and female rats.
No full textThe results of several studies strongly indicate a bidirectional relationship among gonadal hormones and pain. While gonadal hormones play a key role in pain modulation, they have been found to be affected by pain therapies in different experimental and clinical conditions. However, the effects of pain and pain therapy on the gonads are still not clear. In this study, we determined the long-lasting (72 h) effects of inflammatory pain (formalin test) and/or morphine on estrogen receptor (ER), androgen receptor (AR) and TRPV1 gene expression in the rat testis and ovary. The animals were divided into groups: animals receiving no treatment, animals exposed only to the experimental procedure (control group), animals receiving no pain but morphine (sham/morphine), animals receiving pain and morphine (formalin/morphine), and animals receiving only formalin (formalin/saline). Testosterone (T) and estradiol (E) were determined in the plasma at the end of the testing. In the sham/morphine rats, there were increases of ERα, ERβ, AR and TRPV1 mRNA expression in the ovary; in the testis, ERα and ERβ mRNA expression were reduced while AR and TRPV1 expression were unaffected by treatment. T and E plasma levels were increased in morphine-treated female rats, while T levels were greatly reduced in morphine-treated and formalin-treated males. In conclusion, both testicular and ovarian ER (ERα and ERβ) and ovarian AR and TRPV1 gene expression appear to be affected by morphine treatment, suggesting long-lasting interactions among opioids and gonads
Central CRH administration changes formalin pain responses in male and female rats
No full textCorticotropin-releasing hormone (CRH) is suggested to be involved in the regulation of pain. To better evaluate the CRH-mediated behavioral alterations in the formalin inflammatory pain test, we administered CRH or the CRH receptor antagonist α-helical CRH(9-41) (ahCRH) intracerebroventricularly to male and female rats and compared the effects with those of saline control. Nociceptive stimulation was carried out through a subcutaneous injection of dilute formalin (50μL, 10%) in the plantar surface of the hind paw. In both sexes, formalin-induced responses, recorded for 60min, were affected by CRH but not by ahCRH treatment. Paw flexing duration was decreased in both sexes during the formalin interphase period in the CRH-treated group compared to saline control groups; however, licking of the injected paw was markedly increased by the same treatment at other time periods. Treatments induced only a few changes in spontaneous non-pain behaviors, which do not account for the effects on pain response. In conclusion, these data demonstrate the ability of CRH to affect the behavioral responses to an inflammatory nociceptive stimulus, and that the effects can be in opposite directions depending on the behavioral response considere
Gonadal ERα/β, AR and TRPV1 gene expression: Modulation by pain and morphine treatment in male and female rats
No full textThe results of several studies strongly indicate a bidirectional relationship among gonadal hormones and pain. While gonadal hormones play a key role in pain modulation, they have been found to be affected by pain therapies in different experimental and clinical conditions. However, the effects of pain and pain therapy on the gonads are still not clear. In this study, we determined the long-lasting (72. h) effects of inflammatory pain (formalin test) and/or morphine on estrogen receptor (ER), androgen receptor (AR) and TRPV1 gene expression in the rat testis and ovary. The animals were divided into groups: animals receiving no treatment, animals exposed only to the experimental procedure (control group), animals receiving no pain but morphine (sham/morphine), animals receiving pain and morphine (formalin/morphine), and animals receiving only formalin (formalin/saline). Testosterone (T) and estradiol (E) were determined in the plasma at the end of the testing.In the sham/morphine rats, there were increases of ERα, ERβ, AR and TRPV1 mRNA expression in the ovary; in the testis, ERα and ERβ mRNA expression were reduced while AR and TRPV1 expression were unaffected by treatment. T and E plasma levels were increased in morphine-treated female rats, while T levels were greatly reduced in morphine-treated and formalin-treated males.In conclusion, both testicular and ovarian ER (ERα and ERβ) and ovarian AR and TRPV1 gene expression appear to be affected by morphine treatment, suggesting long-lasting interactions among opioids and gonads. © 2012
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