Gonadal ERα/ß, AR and TRPV1 gene expression: modulation by pain and morphine treatment in male and female rats.

The 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, ERß, AR and TRPV1 mRNA expression in the ovary; in the testis, ERα and ERß 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 ERß) 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.

Corticotropin-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 considered.

A new amido phosphonate derivative of carboxymethylcellulose with an osteogenic activity and which is capable of interacting with any Ti surface.

A new phosphonate derivative of carboxymethylcellulose (CMC) was recently synthesized (CMCAPh). The phosphonate polysaccharide was obtained by using a carbodiimide-like activating agent for carboxylic groups and 2-aminoethyl-phosphonic acid to create an amide bond between the amine of the phosphonate agent and the carboxylic acids of CMC. The polymer was characterized by (31)P NMR, FT-IR, and potentiometric titration. CMCAPh showed different properties from CMC and its amidated derivative polymer CMCA. The behavior in solution of CMCAPh polymer towards normal human osteoblasts (NHOst) was studied in vitro, monitoring the cell proliferation, cell differentiation, and osteogenic activity and was then compared with the amidic derivative of carboxymethylcellulose (CMCA). Furthermore, CMCAPh was used to coat titania disks with the aim of increasing the osteogenic activity of implant surfaces. The polymer film on the titania surface was characterized by AFM and TOF-SIMS analysis. An ATR FT-IR study was carried out to evaluate the polymer bonding mode onto the titanium surface. Osteoblast morphology was evaluated by SEM. Adhesion analysis of NHOst demonstrated a better adhesion on the titanium surface coated with CMCAPh than on the bare titanium surface.

Spiral and square microstructured surfaces: the effect of the decreasing size of photo-immobilized hyaluronan domains on cell growth.

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.

Heterotypic cell-cell interaction on micropatterned surfaces.

PURPOSE: The aim of this paper was to study the influence of chemical and topographical signals on cell behavior and to obtain a heterotypic cell-cell interaction on microstructured domains. METHODS: The polysaccharide hyaluronic acid (Hyal) was photoimmobilized on glass surfaces in order to obtain a pattern with squares and rectangles of different dimensions and chemistry. The microstructured surfaces were characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The behavior 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 adherent 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 center to the edge of the sample. 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 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.