Effect of black tea and matrix metalloproteinase inhibitors on eroded dentin in situ

Purpose of this in situ study was to evaluate the surface properties of eroded dentin specimens activated with three different matrix metalloproteinase (MMP) inhibitors (chlorhexidine [CHX], fluoride, green tea), black tea, and water. One hundred eighty dentin samples were prepared from extracted third molars and then samples divided into six groups. Ten volunteers were carried three specimens of each group, on acrylic palatal appliances, which were fabricated exactly for them (n = 3). Erosive cycles were done by immersing appliances in cup containing Cola and was followed by rinsing with test solutions. Microhardness values were measured. Surface properties were investigated by atomic force microscopy (AFM). Lowest change in microhardness was shown in fluoride group whereas negative control group (water) had the highest change. There were no statistically significant differences among surface roughness changes (p > .05). The least change in microhardness was seen in the fluoride group (13.05 ± 8.07), while the control group showed the highest change (33.80 ± 12.42) and was statistically significant when compared to other groups (p < .05). Besides lowest depth, values were shown in fluoride group as well. AFM evaluations showed macromolecular deposits on surfaces of fluoride, CHX, and black tea groups. No superior results were detected in CHX + fluoride group and black tea showed similar surface characteristics as green tea. Mouthrinses containing not only green tea but also black tea could be beneficial for patients with exposed dentin surfaces. Catechines and theaflavins in teas could be useful for improving surface quality.     

Effect of Helicobacter pylori eradication on cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) expression during gastric adaptation to aspirin (ASA) in humans

Gastric adaptation to aspirin is impaired in Helicobacter pylori infection, but the mechanisms underlying this phenomenon are unclear. In this study, we compared gastric mucosal expression of iNOS and COX-2 during 14 days of aspirin ingestion in the same subjects before and 3 months after eradication of H. pylori. Compared to non-infected controls, mucosal expression of COX-2 and iNOS was enhanced before and 3 months after eradication of H. pylori. During aspirin ingestion, mucosal expression of COX-2 remained unchanged before eradication of H. pylori, but increased gradually after successful antimicrobial treatment. Independent of H. pylori status, expression of iNOS increased at the beginning of aspirin intake, but then returned to initial values. We conclude that COX-2 but not iNOS might be involved in gastric adaptation to aspirin in humans and that this mechanism appears to be impaired in H. pylori infection.     

Curcumin protects against hepatic and renal injuries mediated by inducible nitric oxide synthase during selenium‐induced toxicity in Wistar rats

The aim of this study is to evaluate the effect of curcumin in protecting against selenium‐induced toxicity in liver and kidney of Wistar rats. Light microscopy evaluation of selenium alone administered rats showed liver to be infiltrated with mononuclear cells, vacuolation, necrosis, and pronounced degeneration. Control liver sections showed a regular morphology of parenchymal cells with intact hepatocytes and sinusoids. Kidney from selenium alone administered rats showed vacuolar degeneration changes in the epithelial cells, cellular proliferation with fibrosis, thickening of capillary walls, and glomerular tuft atrophy. Such changes were also observed in rats administered with selenium and curcumin simultaneously and rats administered first with selenium and then curcumin 24 h later. Interestingly, such degenerative changes observed in liver and kidney induced by selenium were not seen in rats that were administered with curcumin first and selenium 24 h later. This clearly suggests the protective nature of curcumin against selenium toxicity. To understand the probable mechanism of action of curcumin, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry, and the results showed an increased iNOS expression in selenium‐alone induced liver and kidney. Such high iNOS levels were inhibited in liver and kidney of rats pretreated with curcumin and then with selenium 24 h later. Based on the histological results, it can be concluded that curcumin functions as a protective agent against selenium‐induced toxicity in liver as well as kidney, and this action is probably by the regulatory role of curcumin on iNOS expression. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Immunohistochemical characterization of TH13‐L2 spinal ganglia neurons in sheep (Ovis aries)

Spinal ganglia (SG) neurons are commonly classified according to various specific features. The most widespread classification based on morphological and ultrastructural features subdivides SG neurons into light and small dark neurons. Using immunohistochemical, histochemical and lectin methods, it is possible to further subdivide the small dark neurons into two subpopulations: peptidergic and nonpeptidergic neurons. The majority of studies on SG neurons were carried out on mice and rats; there are few or no studies on large mammals. In this study, some of the widely used neuronal markers, neurofilament 200 kDa (NF200), substance P (SP), calcitonin gene‐related peptide (CGRP) and isolectin B4 (IB4), were employed to characterize neuronal nitric oxide synthase (nNOS)‐immunoreactivity (‐IR) in sheep (Ovis aries) SG (Th13‐L2) neurons. The majority of the SG neurons were IB4‐labeled (79 ± 10%), followed by NF200‐ (45 ± 4%), NOS‐ (44 ± 10%), SP‐ (42 ± 5%) and CGRP‐IR (35 ± 7%) neurons. The triple staining experiments showed that a higher percentage (75 ± 16%) of NOS‐IR neurons bound both IB4 and CGRP, or both IB4 and SP (49 ± 6%). The IB4 marker showed an unexpected staining pattern; in fact, IB4‐labeled neurons largely colocalized with NF200, usually considered a marker of light SG neurons, and with CGRP and SP. For this reason, IB4 cannot be employed in sheep to differentiate between light and dark neurons, or between peptidergic and nonpeptidergic neurons. These results suggest the importance of being cautious when comparing data among different species. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Morphology and lectin‐binding sites of pyloric caeca epithelium in normal and GnRH‐treated Atlantic bluefin tuna (Thunnus thynnus) Linnaeus 1758

Mucosal epithelium of pyloric caeca was studied in normal and in GnRH‐treated Atlantic bluefin tuna Thunnus thynnus L., using morphological analysis, conventional and lectin glycohistochemistry. The lining epithelium consisted of columnar (absorptive) cells, goblet cells and intraepithelial leucocytes. The epithelium from normal animals was significantly taller than GnRH‐treated samples. Conventional histochemistry displayed the same staining pattern in normal and hormone‐treated specimens which showed a mixture of neutral and sulphated acidic glycoconjugates in the luminal surface and goblet cells, and neutral glycans in apical granules of enterocytes. Lectin histochemistry revealed a different glycoconjugate pattern in normal and GnRH‐treated tunas. In normal specimens the luminal surface expressed sialoglycoconjugates which bound MAL II, SNA, KOH‐sialidase‐PNA, KOH‐sialidase‐SBA as well as asialoglycans stained with HPA, SBA, GSA I‐B₄, LTA. N‐linked glycans were highlighted by Con A and KOH‐sialidase‐WGA. In GnRH‐treated tunas the luminal surface did not react with SNA, SBA and LTA. The columnar cells of normal tunas bound KOH‐sialisase‐PNA in the apical region, KOH‐sialidase‐PNA, KOH‐sialidase‐DBA, HPA, SBA, KOH‐sialidase‐SBA and KOH‐sialidase‐WGA in apical granules, GSA I‐B₄ and LTA in the supranuclear region. GnRH‐treated specimens showed some columnar cells that stained with KOH‐sialidase‐WGA in the apical granules and with GSA I‐B₄ in the supranuclear region. The goblet cells of normal animals produced mucins positive to PNA, HPA, KOH‐sialidase‐DBA, SBA, GSA II. The latter three binding sites lacked in GnRH‐treated tunas. The results suggest that the mucosal epithelium of Thunnus thynnus L. pyloric caeca expresses a complex glycan pattern that is affected by GnRH‐treatment. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

The morphological and biomechanical changes of keratocytes cultured on modified p (HEMA‐MMA) hydrogel studied by AFM

The poor integration with host cornea tissue and the low mechanical properties of pHEMA hydrogel for artificial cornea remains a difficult problem to solve. A modified pHEMA hydrogel, MMA copolymerized and type‐I collagen and bFGF immobilized, was previously prepared in an attempt to solve the problems. In this study, the cytotoxicity of Col/bFGF‐p (HEMA‐MMA) and p (HEMA‐MMA) was studied by cell adhesion assay and atomic force microscopy (AFM). The results of cell adhesion assay show that the attachment of keratocytes on the modified membrane is much higher than that of the unmodified membrane. This indicates that the material after modification have better cell–material interaction. The AFM images reveal that the morphology of keratocytes cultured on different substrate is obviously different. The cell cultured on modified membrane presented a completely elongated and spindle‐shape morphology. The force?distance indicates that the biomechanical of keratocytes changes significantly after culturing on different substrates. The adhesion force (2328±523 pN) and Young's modulus (0.51±0.125 kPa) of the cell cultured on modified membrane are much higher, and the stiffness (0.08±0.022 mN/m) is lower than those of the cell cultured on unmodified membrane. These results show that the cytotoxicity of Col/bFGF‐p (HEMA‐MMA) for keratocytes is much improved. SCANNING 31: 246–252, 2009. © 2010 Wiley Periodicals, Inc.     

Morphostructural and immunohistochemical study for evaluation of nano‐TiO2 toxicity in Armadillo officinalis Duméril, 1816 (Crustacea,Isopoda, Oniscidea)

Invertebrates are precious organisms in order to study environmental pollution. In particular, they appear to be suitable as a bioindicator species for pioneer ecotoxicity studies on new xenobiotics such as nanoparticles. In fact, they are able to absorb nanomaterials scattered in the environment in different ways and it's known the compartmentalization of nano‐sized contaminants in selected tissues and intracellular organelles. Titanium dioxide represents the most used nanoparticulate, destined to become probably ubiquitous in the environment. Recently, some research has been published on the toxic potential of nano‐TiO₂ in several animal species. Among all invertebrates, Oniscidean Isopods are the only taxon of Crustaceans that has become completely terrestrial, known as excellent bioindicators and bioaccumulators. They have a digestive gland, the hepatopancreas, which is the location of election for the accumulation of pollutants. For this reason, they are considered efficient animal models to ecological studies. For this study, we collected Armadillo officinalis from Natural Oriented Reserve of “Vendicari” (Sicily, Italy), to evaluate the toxicity of titanium dioxide (TiO₂) on their hepatopancreas, after a short period of exposure. We conducted morphostructural and immunohistochemistry assays. The results suggested a great capacity of the species of bioaccumulation of nanoparticles in the hepatopancreas, where a strong positivity to the metallothioneins was highlighted. Our study confirms that Oniscidean Isopods, in particular Armadillo officinalis, proved to be an appropriate indicator of pollution in terrestrial ecosystems from nanoparticles.     

Effect of long‐term exposure of silver nanoparticles on growth indices,hematological and biochemical parameters and gonad histology of male goldfish (Carassius auratus gibelio)

Studying the impact of emerging pollutants such as nanoparticles is necessary to reveal the adverse effect. In this study, the effects of silver nanoparticles (Ag‐NPs) on hematological, biochemical, and gonad histopathological indices of male goldfish were examined. Sublethal toxicity were calculated based on acute toxicity and three dosages were selected. Live specimen of Carassius auratus gibelio larval were treated in 1, 2, and 3 ppm Ag‐NP with one control group. Blood and tissue samples were extracted after 6 months exposure to sublethal concentrations. Results showed that Ag‐NPs have reduced growth rate and effected on all blood indices significantly. Biochemical analysis revealed that Ag‐NPs significantly reduced blood glucose and total protein than in comparison to the control group and caused significantly differences in the concentrations of serum cholesterol (p < .05). Furthermore, histological observation of intestine after 6 months exposure showed definite alterations in tissue and maximum hypertrophy injuries were found after long‐term exposure to 3 ppm Ag‐NPs concentration. In addition, indicated that long‐term exposure to Ag‐NPs postponed sexual maturity in male gibel carp.