The electric field at the hole-injecting metal/organic interface controls the bias dependence of the current-voltage hole mobility.

Based upon the room temperature current-voltage data of some published organic diode structures the unique phenomenon of thedecreasinghole mobility,µ, with the increasing applied electric field,Ea, is interpreted. The measurable quantity, the hole drift mobilityµdis formulated in terms ofEaand the electric field at the hole injecting metal/organic interface,Eint, dependent algebraic function multiplied by the intrinsic hole mobility,µmaxthat is organic morphology dependent butEaindependent scaling factor. On account that the intrinsic mobility,µmax, is uncoupled from bothEaandEintit is shown that the origin of the negative field hole mobility effect occurs due toEint, that is alinearfunction ofEa. The bias and the space distribution of the internal organic electric field,E, as well as the free hole density,p, for poly(3-hexylthiophene) is calculated in detail. Depending on the organic layer morphology the internal electric field may exhibit, at the particular value ofEa, a deep well in the vicinity of the hole injecting metal/organic interface. Then the strong peak of the free hole density exists there the effect of which is spreading some 10 nm into the organic. IfEinthappens to beEaindependent constant, then from the resulting space charge limited current density, theincreasinghole drift mobility,µd, with the increasing applied electric field,Ea, is deduced. The published current-voltage data of two distinct metal-substituted phthalocyanine thin films provide an additional confirmation of the described formalism.

Enamel Surface Changes After Exposure to Bleaching Gels Containing Carbamide Peroxide or Hydrogen Peroxide.

OBJECTIVE: This study evaluated the differences in enamel color change, surface hardness, elastic modulus, and surface roughness between treatments with four bleaching gels containing carbamide peroxide (two at 10% and one each at 35%, and 45%) and two bleaching gels containing hydrogen peroxide (two at 40%). METHODS: Enamel specimens were bleached and color changes were measured. Color change was calculated using either ΔE or the Bleaching Index (BI). Then, surface hardness, elastic modulus, and surface roughness of the enamel specimens were evaluated. All measurements were performed at baseline and directly after the first bleaching treatment for all carbamide peroxide- and hydrogen peroxide-containing bleaching gels. In addition, final measurements were made 24 hours after each of a total of 10 bleaching treatments for carbamide peroxide bleaching gels, and 1 week after each of a total of three bleaching treatments for hydrogen peroxide bleaching gels. RESULTS: After the last bleaching treatment, respective ΔE scores were 17.6 and 8.2 for the two 10% carbamide peroxide gels, 12.9 and 5.6 for the 45% and 35% carbamide peroxide gels, and 9.6 and 13.9 for the two 40% hydrogen peroxide gels. The respective BI scores were -2.0 and -2.0 for the two 10% carbamide peroxide gels, -3.5 and -1.5 for the 45% and 35% carbamide peroxide gels, and -2.0 and -3.0 for the two 40% hydrogen peroxide gels. Each bleaching gel treatment resulted in significant whitening; however, no significant difference was found among the gels after the last bleaching. Whitening occurred within the first bleaching treatments and did not increase significantly during the remaining treatments. Surface hardness significantly decreased after the last bleaching treatment, when 10% carbamide peroxide was used. Furthermore, significant changes in the elastic modulus or surface roughness occurred only after treatment with 10% carbamide peroxide. CONCLUSION: All six bleaching gels effectively bleached the enamel specimens independent of their concentration of peroxide. Gels with low peroxide concentration and longer contact time negatively affected the enamel surface.

Differential inflammatory response of dental pulp explants and fibroblasts to saliva.

AIM: To investigate the inflammatory response of dental pulp fibroblasts and the respective explants to whole saliva. METHODOLOGY: Explants from human and porcine dental pulp tissue and isolated dental pulp fibroblasts were used to investigate the inflammatory response to sterile saliva. Cytokine and chemokine expression was assessed by RT-PCR. Western blot analysis and pharmacologic inhibitors were used to determine the involvement of signalling pathways. RESULTS: Dental pulp explants of human and porcine origin exposed to human saliva exhibited no major changes of IL-6 and IL-8 mRNA expression (P > 0.05). In contrast, isolated porcine and human dental pulp fibroblasts, when stimulated with human saliva, exhibited a vastly increased expression of IL-6 and IL-8 mRNA (P < 0.05). In pulp fibroblasts, saliva also increased the expression of other cytokines and chemokines via activation of NFkappaB, ERK and p38 signalling. Notably, a significantly reduced inflammatory response was elicited when pulp fibroblasts were transiently exposed to saliva. CONCLUSIONS: Saliva has a potential impact on inflammation of dental pulp fibroblasts in vitro but not when cells are embedded in the intrinsic extracellular matrix of the explant tissue.

Saliva suppresses osteoclastogenesis in murine bone marrow cultures.

Saliva can reach mineralized surfaces in the oral cavity; however, the relationship between saliva and bone resorption is unclear. Herein, we examined whether saliva affects the process of osteoclastogenesis in vitro. We used murine bone marrow cultures to study osteoclast formation. The addition of fresh sterile saliva eliminated the formation of multinucleated cells that stained positive for tartrate-resistant acid phosphatase (TRAP). In line with the histochemical staining, saliva substantially reduced gene expression of cathepsin K, calcitonin receptor, and TRAP. Addition of saliva led to considerably decreased gene expression of receptor activator of nuclear factor kappa-B (RANK) and, to a lesser extent, that of c-fms. The respective master regulators of osteoclastogenesis (c-fos and NFATc1) and the downstream cell fusion genes (DC-STAMP and Atp6v0d2) showed decreased expression after the addition of saliva. Among the costimulatory molecules for osteoclastogenesis, only OSCAR showed decreased expression. In contrast, CD40, CD80, and CD86-all costimulatory molecules of phagocytic cells-were increasingly expressed with saliva. The phagocytic capacity of the cells was confirmed by latex bead ingestion. Based on these in vitro results, it can be concluded that saliva suppresses osteoclastogenesis and leads to the development of a phagocytic cell phenotype.

L-mimosine increases the production of vascular endothelial growth factor in human tooth slice organ culture model.

AIM: To assess the pro-angiogenic and pro-inflammatory capacity of the dentine-pulp complex in response to the prolyl hydroxylase inhibitor L-mimosine in a tooth slice organ culture model. METHODOLOGY: Human teeth were sectioned transversely into 600-µm-thick slices and cultured in medium supplemented with serum and antibiotics. Then, pulps were stimulated for 48 h with L-mimosine. Pulps were subjected to viability measurements based on formazan formation in MTT assays. In addition, histological evaluation of pulps was performed based on haematoxylin and eosin staining. Culture supernatants were subjected to immunoassays for vascular endothelial growth factor (VEGF) to determine the pro-angiogenic capacity and to immunoassays for interleukin (IL)-6 and IL-8 to assess the pro-inflammatory response. Interleukin-1 served as pro-inflammatory control. Echinomycin was used to inhibit hypoxia-inducible factor-1 (HIF-1) alpha activity. Data were analysed using Student's t-test and Mann-Whitney U test. RESULTS: Pulps within tooth slices remained vital upon L-mimosine stimulation as indicated by formazan formation and histological evaluation. L-mimosine increased VEGF production when normalized to formazan formation in the pulp tissue of the tooth slices (P < 0.05). This effect on VEGF was reduced by echinomycin (P < 0.01). Changes in normalized IL-6 and IL-8 levels upon treatment with L-mimosine did not reach the level of significance (P > 0.05), whilst treatment with IL-1, which served as positive control, increased IL-6 (P < 0.05) and IL-8 levels (P < 0.05). CONCLUSIONS: The prolyl hydroxylase inhibitor L-mimosine increased VEGF production via HIF-1 alpha in the tooth slice organ culture model whilst inducing no prominent increase in IL-6 and IL-8. Pre-clinical studies will reveal if these in vitro effects translate into dental pulp regeneration.

The response of dental pulp-derived cells to zoledronate depends on the experimental model.

AIM: To investigate whether zoledronate (ZOL) can cause a cytotoxic response in dental pulp-derived cells (DPCs) in vitro. METHODOLOGY: Cell activity was assessed utilizing MTT tests, (3) [H]thymidine, and (3) [H]leucine incorporation assays in human DPCs in response to ZOL. Cell activity assays were also preformed on calcium phosphate-coated plates. Cell death was analysed with annexin V/propidium iodide, trypan blue staining and Western blot analysis. RESULTS: Micromolar concentrations of ZOL were required to decrease the activity of DPCs. The decreased activity of DPCs was associated with the occurrence of apoptosis and necrosis. No adverse effects were observed when DPCs were cultured on calcium phosphate-coated plates with ZOL. CONCLUSION: High concentrations of soluble ZOL were required to cause adverse effects in vitro. These adverse effects are abolished when the bisphosphonate was bound to a mineralized surface. However, the clinical relevance of these results remains to be determined.

Invitro study of adherent mandibular osteoblast-like cells on carrier materials.

Augmentation of the craniofacial region is necessary for many aesthetic and reconstructive procedures. Tissue engineering offers a new option to supplement existing treatment regimens. In this procedure, materials composed of hydroxyapatite (HA), of synthetic or natural origin, are used as scaffolds. The aim of this study was to evaluate the effects of three HA materials on cultured human osteoblasts in vitro. Explant cultures of cells from human alveolar bone were established. Human osteoblasts were cultured on the surface of HA calcified from red algae (C GRAFT/Algipore), deproteinized bovine HA (Bio-Oss) and bovine HA carrying the cell binding peptide P-15 (Pep Gen P-15). Cultured cells were evaluated with respect to cell attachment, proliferation and differentiation. Cells were cultured for 6 and 21 days under osteogenic differentiation conditions, and tissue-culture polystyrene dishes were used as control. The ability of cells to proliferate and form extracellular matrix on these scaffolds was assessed by a DNA quantification assay, protein synthesis analysis and by scanning electron microscopical examination. Osteogenic differentiation was screened by the expression of alkaline phosphatase. The osteoblastic phenotype of the cells was monitored using mRNA levels of the bone-related proteins including osteocalcin, osteopontin and collagen Type I. We found that cells cultured on C GRAFT/Algipore) and Pep Gen P-15 showed a continuous increase in DNA content and protein synthesis. Cells cultured on Bio-Oss showed a decrease in DNA content from Day 6 (P < 0.05) to Day 21 (P < 0.0001) and protein synthesis on Day 21 (P < 0.005). Alkaline phosphatase activity increased in cells grown on C GRAFT/Algipore and Pep Gen P-15 in contrast to cells grown on Bio-Oss, in which the lowest levels of activity could be observed on Day 21 (P < 0.05). Reverse transcriptase polymerase chain reaction analysis confirmed the osteoblastic phenotype of the cells grown on all three materials throughout the whole culture period. The results of our in vitro study show that the differences in metabolic activity of cells grown on HA materials are directly related to the substrate on which they are grown. They confirm the excellent properties of HA carrying the cell binding peptide P-15 and HA calcified from red algae as used in maxillofacial surgery procedures.

[Evidence of osteocalcin expression in osteoblast cells of mandibular origin growing on biomaterials with RT-PCR and SDS-PAGE/Western blotting]. / Nachweis der Osteokalzinexpression osteoblastärer Zellen mandibulären Ursprungs, wachsend auf Biomaterialien, mittels RT-PCR und SDS-PAGE/Western Blotting.

A new approach to addressing difficult tissue reconstructive or replacement problems in the oral cavity is to engineer new tissue by using selective cell transplantation on polymer scaffolds. The current study characterized the osteoblastic nature of adherent mandibular cells on biomaterials, which could have a potential use as scaffolds for tissue engineering strategies. Cells of mandibular origin from one patient were cultivated on three different biomaterials (PepGen P-15 trade mark, Frios Algipore, and OsteoGraf/LD-700) for 7 and 14 days and osteocalcin expression was demonstrated by RT-PCR and SDS-PAGE/Western blotting. In order to explicitly characterize only the adherent cells on the biomaterials, we first separated the biomaterials with adherent cells from the culture plate before trypsinization. We could demonstrate that cell growth of adherent mandibular osteoblast-like cells was significantly higher on biomaterials with an organic component (PepGen P-15 trade mark ) in comparison to Frios Algipore and OsteoGraf/LD-700, respectively. In conclusion, only the explicit study of adherent cells at the gene and protein levels gives information about the osteoconductivity of biomaterials.