Wael Att

Time-dependent degradation of titanium osteoconductivity: an implication of biological aging of implant materials.

The shelf life of implantable materials has rarely been addressed. We determined whether osteoconductivity of titanium is stable over time. Rat bone marrow-derived osteoblasts were cultured on new titanium disks (immediately after acid-etching), 3-day-old (stored after acid-etching for 3 days in dark ambient conditions), 2-week-old, and 4-week-old disks. Protein adsorption capacity, and osteoblast migration, attachment, spread, proliferation and mineralization decreased substantially on old titanium surfaces in an age-dependent manner. When the 4-week-old implants were placed into rat femurs, the biomechanical strength of bone-titanium integration was less than half that for newly processed implants at the early healing stage. More than 90% of the new implant surface was covered by newly generated bone compared to 58% for 4-week-old implants. This time-dependent biological degradation was also found for machined and sandblasted titanium surfaces and was associated with progressive accumulation of hydrocarbon on titanium surfaces. The new surface could attract osteoblasts even under a protein-free condition, but its high bioactivity was abrogated by masking the surface with anions. These results uncover an aging-like time-dependent biological degradation of titanium surfaces from bioactive to bioinert. We also suggest possible underlying mechanisms for this biological degradation that provide new insights into how we could inadvertently lose, and conversely, maximize the osteoconductivity of titanium-based implant materials.

The effect of UV-photofunctionalization on the time-related bioactivity of titanium and chromium-cobalt alloys.

This study examined the possible changes in the bioactivity of titanium surfaces during their aging and investigated the effect of ultraviolet (UV) light treatment during the age-related change of titanium bioactivity. Rat bone marrow-derived osteoblastic cells were cultured on new titanium disks (immediately after either acid-etching, machining, or sandblasting), 4-week-old disks (stored after processing for 4 weeks in dark ambient conditions), and 4-week-old disks treated with UVA (peak wavelength of 365 nm) or UVC (peak wavelength of 250 nm). During incubation for 24 h, only 50% of the cells were attached to the 4-week-old surfaces as compared to the new surface. UVC treatment of the aged surface increased its cell attachment capacity to a level 50% higher than the new surfaces, whereas UVA treatment had no effect. Proliferation, alkaline phosphatase activity, and mineralization of cells were substantially lower on the 4-week-old surfaces than on the new surfaces, while they were higher on the UVC-treated 4-week-old surfaces as compared to the new surfaces. The age-related impaired bioactivity was found on all titanium topographies as well as on a chromium-cobalt alloy, and was associated with an increased percentage of surface carbon. Although both UVA and UVC treatment converted the 4-week-old titanium surfaces from hydrophobic to superhydrophilic, only UVC treatment effectively reduced the surface carbon to a level equivalent to the new surface. Thus, this study uncovered a time-dependent biological degradation of titanium and chromium-cobalt alloy, and its restoration enabled by UVC phototreatment, which surmounts the innate bioactivity of new surfaces, which is more closely linked to hydrocarbon removal than the induced superhydrophilicity.

Ultraviolet light-mediated photofunctionalization of titanium to promote human mesenchymal stem cell migration, attachment, proliferation and differentiation

Improving the osteoconductive potential of titanium implants has been of continuing interest in the fields of dentistry and orthopedic surgery. This study determined the bioactivity of ultraviolet (UV) light-treated titanium. Human mesenchymal stem cells (MSCs) were cultured on acid-etched microtopographical titanium surfaces with and without 48h pretreatment with UVA (peak wavelength of 360n m) or UVC (peak wavelength of 250 nm). The number of cells that migrated to the UVC-treated surface during the first 3h of incubation was eight times higher than those that migrated to the untreated surface. After 24h of incubation, the number of cells attached to the UVC-treated surface was over three times more than those attached to the untreated surface. On the UVC-treated surface, the cellular spread was expedited with an extensive and intensive expression of the focal adhesion protein vinculin. The cells on the UVC-treated surface exhibited a threefold higher bromodeoxyuridine incorporation, a doubling of the alkaline phosphatase-positive area and the up-regulated expression of bone-related genes, indicating the accelerated proliferation and differentiation. The UVC-treated surface did not adversely affect the viability of the cells. These biological effects were not seen after UVA treatment, despite the generation of superhydrophilicity. Thus, we discovered a novel photofunctionalization of titanium dioxide that substantially enhances its bioactivity in human MSCs. Further studies are required to investigate the universal effectiveness of this surface modification for different titanium-containing materials, with varying chemistries and textures, as well as to understand its significance in enhancing in vivo osteoconductivity.

Marginal adaptation of three different zirconium dioxide three-unit fixed dental prostheses

STATEMENT OF PROBLEMnMarginal adaptation is important for the long-term success of dental restorations. Data on the marginal discrepancy of zirconia-based fixed dental prostheses made with different computer-aided design/computer-aided manufacturing technology is needed.nnnPURPOSEnThe purpose of this study was to evaluate the marginal adaptation of different zirconia 3-unit fixed dental prostheses at different fabrication stages and after artificial aging.nnnMATERIAL AND METHODSnTwenty-four zirconia 3-unit fixed dental prostheses (DCS, Procera, and VITA YZ-Cerec; n=8) were fabricated using different manufacturing systems and conventionally cemented with glass ionomer cement on human teeth. Each group was aged in a masticatory simulator with thermal cycling. The marginal gaps were examined on epoxy replicas for frameworks and for restorations before and after cementation, and after masticatory simulation, at x 250 magnification. Marginal adaptation was assessed using geometric means of the marginal gap values with 95% confidence intervals. Differences between the manufacturing systems and the effect of artificial aging were tested using repeated-measures ANOVA and post hoc paired and unpaired t tests with Bonferroni-Holm correction (alpha=.05).nnnRESULTSnThe geometric mean (95% confidence limits) marginal gap values (mum) for frameworks and for restorations before cementation, after cementation, and after masticatory simulation were, respectively: DCS: 86 (80-93), 86 (83-90), 86 (78-94), and 84 (79-90); Procera: 82 (74-89), 89 (81-97), 89 (84-95), and 88 (82-94); and VITA YZ-Cerec: 64 (57-72), 67 (61-77), 76 (71-82), and 78 (76-80). The repeated-measures ANOVA showed significant group and stage effects (P<.05). Group VITA YZ-Cerec showed significantly smaller marginal gap values than groups DCS and Procera at framework (P<.05) and before-cementation (P<.05) stages. The VITA YZ-Cerec group showed significantly smaller marginal gap values than the Procera group after cementation (P<.05). The marginal gap values between different stages were not significantly different for all groups (P>.05).nnnCONCLUSIONSnThe marginal accuracy of zirconia fixed dental prostheses is influenced by manufacturing technique.

Age-dependent Degradation of the Protein Adsorption Capacity of Titanium:

Reported bone-implant contact percentages are far below the ideal 100%. We tested a hypothesis that the protein adsorption capability of titanium, which is critical to the process of osseointegration, changes over time before its use. Machined, acid-etched, and sandblasted surfaces were prepared and stored under dark ambient conditions for 3 days, 1 week, or 4 weeks. For all surfaces, protein adsorption decreased as the storage time increased, and their decreasing rates were dependent on titanium topography. After 4 weeks, the amounts of albumin and fibronectin adsorbed by the acid-etched surface were only 20% and 35%, respectively, of that adsorbed by the fresh surface after 2 hours of incubation, and remained substantially low even after 24 hours. This time-dependent degradation in protein adsorption of titanium correlated with its naturally decreasing hydrophilicity, which was not observed for the nickel and chromium surfaces, indicating a titanium-specific biological aging.

Enhanced osteoblast function on ultraviolet light-treated zirconia

Unlike titanium, surface roughening of zirconia for enhanced bone integration has been technically challenging. The photochemical reaction of semiconductor oxides, e.g., titanium dioxide, has earned considerable and broad interest in environmental and clean energy sciences. This study determined whether ultraviolet (UV) light treatment of zirconia enhances its bioactivity on osteoblasts. Machined zirconia disks were treated with UV light for various time periods up to 48 h. UV light treatment for 48 h increased the rates of attachment, spread, and proliferation of rat bone marrow-derived osteoblasts. Alkaline phosphatase-positive and mineralized nodule areas doubled on UV light-treated zirconia. The expression of osteoblastic genes, such as osteopontin and osteocalcin, was not modulated by UV light treatment. X-ray diffraction and X-ray photoelectron spectroscopy analyses showed that zirconia disks consisted of monoclinic and tetragonal phases of ZrO(2) and Y(2)O(3) having a wide light absorption band of 200-400 nm with its peak at <250 nm. UV light treatment transformed the zirconia surface from hydrophobic to hydrophilic status and reduced the atomic percentage of surface carbon in a UV light dose-dependent manner. These results suggest that UV treatment of yttrium-containing partially stabilized zirconia enhances its bioactivity on osteoblasts, in terms of their attachment, proliferation, and eventually mineralization. This biofunctionalization was associated with UV light-catalytic hydrophilic conversion of zirconia surfaces and progressive removal of hydrocarbons.

Fixed Rehabilitation of the Edentulous Maxilla: Possibilities and Clinical Outcome

PURPOSEnThe aim of the present report was to describe the different treatment approaches available for fixed rehabilitation of the edentulous maxilla in the presence of varying hard and soft tissue conditions and to review the clinical outcome of each treatment approach.nnnMATERIALS AND METHODSnA review of the published data published from 1980 through 2009 was conducted using electronic databases and manual searching to identify the treatment possibilities for the fixed rehabilitation of the edentulous maxilla and report their clinical outcomes. The search terms used, in simple or multiple conjunctions, were fixed rehabilitation, implants, edentulous, fixed dental prosthesis, implant-supported, and maxilla.nnnRESULTSnSeveral treatment modalities were identified for the fixed rehabilitation of the edentulous maxilla, with and without bone augmentation procedures. Regular, tilted, and zygoma implants were identified for treatment modalities that do not require bone augmentation. Sinus floor elevation with the lateral window technique or Le Fort I osteotomy with interpositional bone grafts was identified as a treatment possibility that required bone augmentation procedures. The database initially yielded 230 titles. Of the 230 studies, 42 were finally selected. Although all studies reported the survival rates of the implants, only 20 provided information about the prosthetic outcome. Because of the limited number of studies, at least for the specific treatment modalities, and the heterogeneity in the design of the different studies identified, it was not possible to perform a statistical analysis of the data. Except for regular implants placed in native bone, no sufficient long-term clinical studies were found for the other procedures.nnnCONCLUSIONSnExcept for regular implants placed in nonaugmented native bone, the published data provide insufficient evidence about the outcome of other procedures. Until long-term data are available, such procedures should not be considered reliable treatment modalities.

N-Acetyl cysteine prevents suppression of oral fibroblast function on poly(methylmethacrylate) resin

Despite the proven cytotoxicity, poly(methylmethacrylate) (PMMA) resin is one of the most frequently and extensively used materials in medical and dental fields. The study examined the potential detoxification of the resin material and restoration of the resin-induced suppression of cellular function using an antioxidant amino acid derivative, N-acetyl cysteine (NAC). Oral fibroblasts extracted from rat oral mucosa were cultured on the resin material with or without incorporation of NAC into the material. Twenty-four hour after incubation, less than 2% of the cells were viable on the untreated control resin, while up to 35% of the cells were viable on the resin with incorporation of NAC. At day 7 of culture, the expression of collagen I and III genes was downregulated on the untreated resin, while the cells on NAC-supplemented resin showed the expression levels similar to those in polystyrene culture. The cells produced three times greater amount of collagen on the NAC-supplemented resin than on the untreated resin. The data demonstrated that the cytotoxicity of PMMA resin was substantially lower when the material contains NAC. The potential usefulness of this principle should be explored with a view of developing biocompatible polymer-based materials in a broad range of dental and medical resin materials and tissue engineering scaffolds.

N-Acetyl cysteine (NAC) inhibits proliferation, collagen gene transcription, and redox stress in rat palatal mucosal cells

OBJECTIVESnControl of hyperplastic and invasively growing gingival tissue is crucial for maintaining normal oral function and for successful bone regenerative therapy. We tested the hypothesis that materials containing N-acetyl cysteine (NAC), an antioxidant cysteine derivative, can control proliferation and function of oral mucosal cells.nnnMETHODSnOral mucosal cells derived from the rat palatal tissue were cultured with or without NAC at different concentrations (2.5-10.0mM). To simulate inflammatory conditions, cultures were treated with hydrogen peroxide. NAC was also applied via collagen materials in membrane and sponge forms to explore the clinical applicability. The redox balance inside the cells was evaluated by measuring the concentration of intracellular glutathione (GSH).nnnRESULTSnAdding NAC into cultures of oral mucosal cells reduced their proliferation, transcriptional expression, and collagen production in an NAC-concentration-dependent manner without cytotoxic effects. Furthermore, NAC substantially reduced the hydrogen peroxide-induced elevation of cellular proliferation and collagen production. The controlling effects of NAC were also demonstrated in cells cultured on NAC-containing collagen materials and were associated with an increase in intracellular glutathione (GSH) reserves and a decrease in the oxidized form of glutathione (GSSG).nnnSIGNIFICANCEnThese results indicate that NAC may abrogate inflammation- or oxidative-stress-induced hyperfunction of oral mucosal cells and that it can be delivered effectively via biodegradable materials. This study provides a basis to explore NAC-containing biomaterials that are functionalized to control oral soft tissue growth and function without cytotoxicity.

N-Acetyl cysteine restores viability and function of rat odontoblast-like cells impaired by polymethylmethacrylate dental resin extract

Abstract There is concern that dental-resin materials directly loaded on a prepared tooth adversely affect dental pulp tissue by releasing the resin chemicals through dentinal tubes. This study determined whether self-curing polymethyl methacrylate (PMMA)-based dental resin extract adversely affected the viability and function of odontoblast-like cells and whether the cytotoxicity of this resin, if any, could be eliminated by N-acetyl cysteine, an antioxidant amino acid derivative. Odontoblast-like cells isolated from rat maxillary incisor dental pulp tissue were exposed to a PMMA resin extract with or without N-acetyl cysteine for 1 h and then cultured in osteoblastic media. The percentage of viable cells 24 h after seeding was 20% in cells exposed to the resin extract without N-acetyl cysteine, whereas 45% of cells were viable after exposure to the N-acetyl cysteine-supplemented extract. The cells that had been exposed to the extract showed a strong tendency for apoptosis associated with the increased reactive oxygen species production and decreased intracellular glutathione level, which was improved by the addition of N-acetyl cysteine. N-Acetyl cysteine supplementation almost completely restored the significantly reduced alkaline phosphatase activity and matrix mineralization by the resin extract. These results conclusively demonstrated that exposure of odontoblast-like cells to the resin extract impaired the cell viability and function and, more intriguingly, N-acetyl cysteine supplementation to the extract significantly prevented these toxic effects.