The ARTUS device: the first feasibility study in human cadavers.

INTRODUCTION: The aim was to perform a feasibility study of the new artificial sphincter device ARTUS in human cadavers. ARTUS is a new electro-mechanical device, which may prevent urethral damage due to a new working principle which is to perform only sequential pressure on successive parts of the urethra. MATERIAL AND METHODS: The implantation of the ARTUS device was performed in six cadavers (3 males, 3 females) with different body mass indices. Subsequently the basic operation data (operation time, cuff size, length of wires, complication) were assessed. RESULTS: The implantation of the ARTUS device is performed easily by the same technique which is commonly used for the AMS 800 implantation. The mean operation time was 20 minutes. The mean cuff size was 4.5 cm in male and 6 cm in female cadavers. The average length of the wires was 12 cm. The necessary subcutaneous pouch had to be bigger than the space used for the tubes of the AMS 800 device. The study is limited by its preclinical setting. CONCLUSIONS: Our results demonstrate that this new artificial urinary sphincter device can be easily implanted. The technical and surgical approaches are similar to those which are applied in the case of the AMS 800 device. Therefore experienced surgeons will be able to adapt their technique easily.

Polyspecies biofilm formation on implant surfaces with different surface characteristics.

OBJECTIVE: To investigate the microbial adherence and colonization of a polyspecies biofilm on 7 differently processed titanium surfaces. MATERIAL AND METHODS: Six-species biofilms were formed anaerobically on 5-mm-diameter sterilized, saliva-preconditioned titanium discs. Material surfaces used were either machined, stained, acid-etched or sandblasted/acid-etched (SLA). Samples of the latter two materials were also provided in a chemically modified form, with increased wettability characteristics. Surface roughness and contact angles of all materials were determined. The discs were then incubated anaerobically for up to 16.5 h. Initial microbial adherence was evaluated after 20 min incubation and further colonization after 2, 4, 8, and 16.5 h using non-selective and selective culture techniques. Results at different time points were compared using ANOVA and Scheffé post hoc analysis. RESULTS: The mean differences in microorganisms colonizing after the first 20 min were in a very narrow range (4.5 to 4.8 log CFU). At up to 16.5 h, the modified SLA surface exhibited the highest values for colonization (6.9±0.2 log CFU, p<0.05) but increasing growth was observed on all test surfaces over time. Discrepancies among bacterial strains on the differently crafted titanium surfaces were very similar to those described for total log CFU. F. nucleatum was below the detection limit on all surfaces after 4 h. CONCLUSION: Within the limitations of this in vitro study, surface roughness had a moderate influence on biofilm formation, while wettability did not seem to influence biofilm formation under the experimental conditions described. The modified SLA surface showed the highest trend for bacterial colonization.

Submicron scale-structured hydrophilic titanium surfaces promote early osteogenic gene response for cell adhesion and cell differentiation.

BACKGROUND AND PURPOSE: Titanium (Ti) surface roughness and surface hydrophilicity are key factors to regulate osteogenic cell responses during dental implant healing. In detail, specific integrin-mediated interactions with the extracellular environment trigger relevant osteogenic cell responses like differentiation and matrix synthesis via transcriptions factors. Aim of this study was to monitor surface-dependent osteogenic cell adhesion dynamics, proliferation, and specific osteogenic cell differentiation over a period of 7 days. MATERIALS AND METHODS: Ti disks were manufactured to present smooth pretreatment (PT) surfaces and rough sandblasted/acid-etched (SLA) surfaces. Further processing to isolate the uncontaminated TiO(2) surface from contact with atmosphere provided a highly hydrophilic surface without alteration of the surface topography (modSLA). Tissue culture polystyrene (TCPS) served as control. Human osteogenic cells were cultivated on the respective substrates. After 24 hours, 48 hours, 72 hours, and 7 days, cell morphology on the Ti substrates was visualized by scanning transmission electron microscopy. As a marker of cellular proliferation, cell count was assessed. For the analysis of cell adhesion and differentiation, specific gene expression levels of the integrin subunits ß1 and αv, runx-2, collagen type Iα (COL), alkaline phosphatase (AP), and osteocalcin (OC) were obtained by real-time RT-PCR for the respective time points. Data were normalized to internal controls. RESULTS: TCPS and PT surfaces preserved a rather immature, dividing osteogenic phenotype (high proliferation rates, low integrin levels, and low specific osteogenic cell differentiation). SLA and especially modSLA surfaces promoted both cell adhesion as well as the maturation of osteogenic precursors into post-mitotic osteoblasts. In detail, during the first 48 hours, modSLA resulted in lowest cell proliferation rates but exhibited highest levels of the investigated integrins, runx-2, COL, AP, and OC. CONCLUSION: Our results revealed a strong synergistic effect between submicron-scale roughness and surface hydrophilicity on early osteogenic cell adhesion and maturation.

Polyspecies biofilm formation on implant surfaces with different surface characteristics

Objective: To investigate the microbial adherence and colonization of a polyspecies biofilm on 7 differently processed titanium surfaces. Material and Methods: Six-species biofilms were formed anaerobically on 5-mm-diameter sterilized, saliva-preconditioned titanium discs. Material surfaces used were either machined, stained, acid-etched or sandblasted/acid-etched (SLA). Samples of the latter two materials were also provided in a chemically modified form, with increased wettability characteristics. Surface roughness and contact angles of all materials were determined. The discs were then incubated anaerobically for up to 16.5 h. Initial microbial adherence was evaluated after 20 min incubation and further colonization after 2, 4, 8, and 16.5 h using non-selective and selective culture techniques. Results at different time points were compared using ANOVA and Scheffé post hoc analysis. Results: The mean differences in microorganisms colonizing after the first 20 min were in a very narrow range (4.5 to 4.8 log CFU). At up to 16.5 h, the modified SLA surface exhibited the highest values for colonization (6.9±0.2 log CFU, p<0.05) but increasing growth was observed on all test surfaces over time. Discrepancies among bacterial strains on the differently crafted titanium surfaces were very similar to those described for total log CFU. F. nucleatum was below the detection limit on all surfaces after 4 h. Conclusion: Within the limitations of this in vitro study, surface roughness had a moderate influence on biofilm formation, while wettability did not seem to influence biofilm formation under the experimental conditions described. The modified SLA surface showed the highest trend for bacterial colonization.

Initial attachment, subsequent cell proliferation/viability and gene expression of epithelial cells related to attachment and wound healing in response to different titanium surfaces.

OBJECTIVES: A tight seal between the epithelium and the dental implant surface is required to prevent bacterial inflammation and soft tissue recession and therefore to demonstrate a long-term success. Surface hydrophilicity was recently shown to promote osseointegration. The aim of this study was to investigate the influence of surface hydrophilicity in combination with surface topography of Ti implant surfaces on the behavior and activation/differentiation of epithelial cells using a set of in vitro experiments mimicking the implant-soft tissue contact. METHODS: Hydrophobic acid-etched (A) and coarse-grit-blasted, acid-etched (SLA) surfaces and hydrophilic acid-etched (modA) and modSLA surfaces were produced. The behavior of an oral squamous cell carcinoma cell line (HSC-2) grown on all surfaces was compared through determination of cell attachment and proliferation/viability (CCK-8 and MTT assay), time-lapse microscopy of fluorescence labeled cells and determination of gene expression by real time polymerase chain reaction. RESULTS: Within the surfaces with similar wettability cell spreading and cell movements observed by time-lapse microscopy after one day of incubation were most pronounced on smoother (A and modA) surfaces compared to rougher (SLA and modSLA) surfaces. Within the surfaces with similar roughness the hydrophilic surfaces (modA and modSLA) showed more cell spreading and cell activity compared to the hydrophobic surfaces (A and SLA). The relative gene expressions of cytokeratin14, integrin α6, integrin ß4, vinculin, transforming growth factor (TGF)-ß, TGF-ß1, and TGF-ß3 were decreased in HSC-2 on all four types of Ti surfaces compared to control surfaces (tissue culture polystyrene; p<0.01) and there was no significant difference of gene expression on the four different implant-surfaces. SIGNIFICANCE: We have demonstrated that for proliferation and spreading of HSC-2 cells the smoother and hydrophilic surface is optimal (modA). These results suggest that surface hydrophilicity might positively influence the epithelial seal around dental implants. All tested titanium surfaces downregulate cell attachment, cell proliferation, expression of adhesion promoters, and cytokines involved in wound healing in HSC-2 cells compared to control surfaces.

Biomechanical evaluation of a microstructured zirconia implant by a removal torque comparison with a standard Ti-SLA implant.

OBJECTIVES: The purpose of this study was to evaluate the biomechanical bone tissue response to novel microstructured zirconia implants in comparison to sandblasted and acid-etched (SLA) titanium implants through the analysis of removal torque (RTQ) measurements. MATERIALS AND METHODS: Ti-SLA implants with a sandblasted, large-grit and acid-etched surface were compared with zirconia implants with an acid-etched surface. All implants had the same shape, a diameter of 4.1 mm and a length of 10 mm. A total of 136 implants were placed in the maxillae of 17 miniature pigs. Six animals were sacrificed after both 4 and 8 weeks and five animals were sacrificed after 12 weeks, thus providing a total of 102 implants for RTQ testing (34 implants were reserved for future histological analysis). The RTQ analysis was successfully performed, using a mixed model regression with P-values calculated using the nonparametric Brunner-Langer method, on 100 of the 102 implants, two were excluded from the analysis. RESULTS: The adjusted mean RTQ values for Ti-SLA implants were 131 Ncm (95% CI: 107-155) at 4 weeks, 128 Ncm (108-148) Ncm at 8 weeks, and 180 Ncm (153-207 Ncm) at 12 weeks of healing, whereas RTQ values for the zirconia implants were 110 Ncm (86-134), 97 Ncm (76-118) and 147 Ncm (121-174) at the corresponding time intervals. A comparison of the implant materials resulted in P-values of P = 0.114 at 4 weeks, P = 0.034 at 8 weeks and P = 0.105 at 12 weeks (significance set at P < 0.05). CONCLUSIONS: Within the limits of the present study, it could be confirmed that the biomechanical bone-tissue response of the investigated zirconia implants is non-inferior to that of the well-documented, roughened titanium surface, at each time point, within the set tolerance. There were no statistically significant differences between the two materials after a healing period of 4 and 12 weeks. The RTQ values of both implant types increased significantly from the 8-week to the 12-week time point.

Primary stability of a hybrid self-tapping implant compared to a cylindrical non-self-tapping implant with respect to drilling protocols in an ex vivo model.

BACKGROUND: Modifications of implant design have been intending to improve primary stability. However, little is known about investigation of a hybrid self-tapping implant on primary stability. PURPOSES: The aims of this study were to evaluate the primary stability of two hybrid self-tapping implants compared to one cylindrical non-self-tapping implant, and to elucidate the relevance of drilling protocols on primary stability in an ex vivo model. MATERIALS AND METHODS: Two types of hybrid self-tapping implants (Straumann® Bone Level implant [BL], Straumann® Tapered Effect implant [TE]) and one type of cylindrical non-self-tapping implant (Straumann® Standard Plus implant [SP]) were investigated in the study. In porcine iliac cancellous bones, 10 implants each were inserted either using standard drilling or under-dimensioned drilling protocol. The evaluation of implant-bone interface stability was carried out by records of maximum insertion torque, the Periotest® (Siemens, Bensheim, Germany), the resonance frequency analysis (RFA), and the push-out test. RESULTS: In each drilling group, the maximum insertion torque values of BL and TE were significantly higher than SP (p=.014 and p=.047, respectively). In each group, the Periotest values of TE were significantly lower than SP (p=.036 and p=.033, respectively). The Periotest values of BL and TE were significantly lower in the group of under-dimensioned drilling than standard drilling (p=.002 and p=.02, respectively). In the RFA, no statistical significances were found in implants between two groups and between implants in each group. In each group, the push-out values of BL and TE were significantly higher than SP (p=.006 and p=.049, respectively). CONCLUSION: Hybrid self-tapping implants could achieve a high primary stability which predicts them for use in low-density bone. However, there is still a debate to clarify the influence of under-dimensioned drilling on primary stability.

Osteogenic properties of hydrophilic and hydrophobic titanium surfaces evaluated with osteoblast-like cells (MG63) in coculture with human umbilical vein endothelial cells (HUVEC).

OBJECTIVES: Osteogenesis on titanium (Ti) surfaces is a complex process involving cell-substrate and cell-cell interaction of osteoblasts and endothelial cells. The aim of this study was to investigate the osteogenic properties of Ti surfaces on osteoblasts in the presence of endothelial cells (ECs). METHODS: Osteoblast-like cells (MG63 cells) and human umbilical vein endothelial cells (HUVECs) were grown in cocultures on four kinds of Ti surfaces: acid-etched (A), coarse-grit-blasted and acid-etched (SLA), hydrophilic A (modA) and hydrophilic SLA (modSLA) surfaces. MG63 cells in single cultures served as controls. Cell ratios and cell types in cocultures were determined and isolated using flow cytometry. Cell numbers were obtained by direct cell counting. In MG63 cells, alkaline phosphatase (ALP) activity was determined and protein levels of osteocalcin (OC) and osteoprotegerin (OPG) were detected with enzyme-linked immunosorbant assay (ELISA). The mRNA levels of ALP, OC and OPG of sorted MG63 cells were determined with real time polymerase chain reaction (PCR). RESULTS: MG63 cells proliferated in the presence of HUVECs, which showed higher cell numbers on Ti surfaces (A, SLA, modSLA) after 72h, and lower cell numbers on Ti surfaces (modA, SLA, modSLA) after 120h in comparison to single cultures. Protein and mRNA levels of ALP and OPG were higher in cocultures than in single cultures, while OC exhibited a lower expression. These three parameters were higher expressed on modA, SLA and modSLA surfaces compared to A surfaces. SIGNIFICANCE: Cocultures of osteoblasts and endothelial cells represent the most recently developed research model for investigating osteogenesis and angiogenesis which play both a major role in bone healing. This paper investigates for the first time the osteogenic properties of titanium surfaces used for dental implants with a coculture system with osteoblast-like cells and endothelial cells: (1) In cocultures with ECs (HUVECs) osteoblast-like cells (MG63 cells) show enhanced expression of early differentiation markers and osteogenic factors on Ti surfaces compared to single cultures of MG63 cells. (2) The differentiation and the expression of an osteogenic phenotype of osteoblast-like cells (MG63 cells) in coculture with ECs (HUVECs) is enhanced by both hydrophilicity and roughness of Ti surfaces.

Impact of guided bone regeneration and defect dimension on wound healing at chemically modified hydrophilic titanium implant surfaces: an experimental study in dogs.

OBJECTIVES: The aim of the present study was to evaluate the impact of guided bone regeneration and defect dimension on wound healing at chemically modified titanium implant surfaces (modSLA). MATERIALS AND METHODS: ModSLA implants were placed at chronic-type lateral ridge defects of different heights (H1-H4: 2, 4, 6 and 8 mm) and randomly allocated to either (a) GBR (polyethylene glycol membrane + biphasic calcium phosphate) or (b) untreated control. At 2 and 8 weeks (n=6 dogs each), dissected blocks were processed for histomorphometrical analysis [e.g., percentage linear fill (PLF), regenerated area (RA)]. RESULTS: At 8 weeks, both groups revealed comparable mean PLF (%) [ CONTROL: H1 (26.1 +/- 5.8)-H4 (60.4 +/- 11.8); GBR: H1 (8.3 +/- 5.3)-H4 (50.7 +/- 23.1)] and RA (mm(2)) [ CONTROL: H1 (2.5 +/- 0.4)-H4 (7.4 +/- 4.1); GBR: H1 (1.8 +/- 1.0)-H4 (10.8 +/- 5.9)] values. A significant difference was observed for the mean PLF values at H1 defects. CONCLUSION: It was concluded that (i) modSLA titanium implants supported bone regeneration and osseointegration at H1-H4 defects and (ii) the present GBR procedure did not seem to improve the outcome of vertical bone regeneration, but tended to increase the mean RA values.

Influence of frequent clinical probing during the healing phase on healthy peri-implant soft tissue formed at different titanium implant surfaces: a histomorphometrical study in dogs.

OBJECTIVES: To investigate (i) the impact of different titanium implant surfaces on soft tissue integration over 6 months, and (ii) the influence of frequent clinical probing during the healing phase on the established mucosal seal. MATERIAL AND METHODS: Standardized clinical probing was randomly performed (12 dogs, probing versus control) at different transmucosal surfaces [machined (M), sand-blasted/acid-etched (SLA), and chemically modified acid-etched (modA), modSLA] at 2, 4, 8, and 12 weeks (i.e. 1 x , 2 x , 3 x , and 4 x). Histomorphometrical analysis (e.g. mucosal margin (PM) - apical extension of the junctional epithelium (aJE), PM - coronal level of bone-to-implant contact (CBI) was performed at 4, 8, 12, and 24 weeks. RESULTS: While M and SLA groups revealed a split formation, epithelial cells and connective tissue were in close contact to modA and modSLA surfaces. Frequent clinical probing (i.e. 3 x and 4 x) increased mean pocket depths, PM-aJE, and aJE-CBI values in all groups and markedly disrupted the epithelial and connective tissue attachment. CONCLUSIONS: It was concluded that irrespective of the surface characteristics, a frequent clinical probing at short intervals during the healing phase was associated with dimensional and structural changes of the mucosal seal.

Regulation of angiogenesis during osseointegration by titanium surface microstructure and energy.

Rough titanium (Ti) surface microarchitecture and high surface energy have been shown to increase osteoblast differentiation, and this response occurs through signaling via the alpha(2)beta(1) integrin. However, clinical success of implanted materials is dependent not only upon osseointegration but also on neovascularization in the peri-implant bone. Here we tested the hypothesis that Ti surface microtopography and energy interact via alpha(2)beta(1) signaling to regulate the expression of angiogenic growth factors. Primary human osteoblasts (HOB), MG63 cells and MG63 cells silenced for alpha(2) integrin were cultured on Ti disks with different surface microtopographies and energies. Secreted levels of vascular endothelial growth factor-A (VEGF-A), basic fibroblast growth factor (FGF-2), epidermal growth factor (EGF), and angiopoietin-1 (Ang-1) were measured. VEGF-A increased 170% and 250% in MG63 cultures, and 178% and 435% in HOB cultures on SLA and modSLA substrates, respectively. In MG63 cultures, FGF-2 levels increased 20 and 40-fold while EGF increased 4 and 6-fold on SLA and modSLA surfaces. These factors were undetectable in HOB cultures. Ang-1 levels were unchanged on all surfaces.Media from modSLA MG63 cultures induced more rapid differentiation of endothelial cells and this effect was inhibited by anti-VEGF-A antibodies. Treatment of MG63 cells with 1 alpha,25(OH)(2)D3 enhanced levels of VEGF-A on SLA and modSLA.Silencing the alpha(2) integrin subunit increased VEGF-A levels and decreased FGF-2 levels. These results show that Ti surface microtopography and energy modulate secretion of angiogenic growth factors by osteoblasts and that this regulation is mediated at least partially via alpha(2)beta(1) integrin signaling.

Effect of different surface pre-treatments and luting materials on shear bond strength to PEEK.

OBJECTIVES: To assess the bonding potential of a universal composite resin cement and an adhesive/composite system to differently pre-treated PEEK surfaces. METHODS: One hundred and fifty PEEK disks were embedded in epoxy resin, polished (P4000 grit) and treated as follows (n=30/group): (A) no treatment, (B) acid etching with sulfuric acid (98%) for 1 min, (C) sandblasting for 10s with 50 microm alumina, (D) sandblasting for 10s with 110 microm alumina and (E) silica coating using the Rocatec system (3M ESPE). Polished and sandblasted (50 microm alumina) cp titanium (grade 4) served as a control. Acrylic hollow cylinders were either luted with a universal composite resin cement (RelyX Unicem) or an unfilled resin (Heliobond) and a hybrid composite (Tetric) to the specimens. Bond strength was measured in a shear test and failure modes were assessed. Statistic analysis was performed with one-way ANOVA followed by a post hoc Scheffé test and unpaired t-tests. RESULTS: With the universal composite resin cement, no bond could be established on any PEEK surfaces, except specimens etched with sulfuric acid (19.0+/-3.4MPa). Shear bond strength to titanium was significantly lower (8.7+/-2.8MPa, p<0.05). Applying the adhesive/composite system, shear bond strength values on pre-treated PEEK ranged from 11.5+/-3.2MPa (silica coating) to 18.2+/-5.4MPa (acid etched) with no statistically significant differences (p>0.05). No bond was obtained on the polished surface. SIGNIFICANCE: Bonding to PEEK is possible when using a bonding system. No adherence can be achieved with the tested universal composite resin cement except on an etched surface. The results strongly encourage further research in PEEK application in dentistry.

Surface alterations of polished and sandblasted and acid-etched titanium implants after Er:YAG, carbon dioxide, and diode laser irradiation.

PURPOSE: Laser treatment has become a popular method for resolving peri-implantitis, but the full range of its effects on implant surfaces is unknown. The purpose of the present investigation was to analyze the influence of different clinically applicable erbium:yttrium-aluminum-garnet (Er:YAG), carbon dioxide (CO2), and diode laser parameters on titanium surfaces that were either polished or sandblasted, large-grit, acid-etched (SLA). MATERIALS AND METHODS: Six polished and six SLA titanium disks were irradiated at nine different power settings (n = 54 polished, 54 SLA) with Er:YAG, CO2, or diode lasers. The CO2 and diode lasers were used in continuous wave mode, and the Er:YAG laser was used in a pulsed manner. The surface of each disk was analyzed by scanning electron microscopy and confocal white light microscopy. Each disk was irradiated on six circular areas of 5 mm in diameter with the same specific laser setting for 10 seconds. RESULTS: Within the chosen parameters, the CO2 and diode laser did not cause any visible surface alterations on either the polished or SLA disks. In contrast, both polished and SLA disks showed surface alterations when irradiated with the pulsed Er:YAG laser. The SLA surfaces showed alteration after 10 seconds of irradiation with Er:YAG laser at 300 mJ/10 Hz. The surfaces of the polished disks did not show alteration with the Er:YAG laser until they were irradiated at the higher energy of 500 mJ/10 Hz for 10 seconds. The results of confocal white light microscopy were in agreement with scanning electron micrographs. CONCLUSION: In contrast to continuous-wave diode and CO2 laser irradiation, pulsed Er:YAG laser irradiation caused distinct alterations with power settings beyond 300 mJ/10 Hz on the SLA surface and 500 mJ/10 Hz on the polished surface. Thus, it is only safe to use the Er:YAG laser for implant surface irradiation with settings no higher than 300 or 500 mJ/10 Hz.

Direct and indirect effects of microstructured titanium substrates on the induction of mesenchymal stem cell differentiation towards the osteoblast lineage.

Microstructured and high surface energy titanium substrates increase osseointegration in vivo. In vitro, osteoblast differentiation is increased, but effects of the surface directly on multipotent mesenchymal stem cells (MSCs) and consequences for MSCs in the peri-implant environment are not known. We evaluated responses of human MSCs to substrate surface properties and examined the underlying mechanisms involved. MSCs exhibited osteoblast characteristics (alkaline phosphatase, RUNX2, and osteocalcin) when grown on microstructured Ti; this effect was more robust with increased hydrophilicity. Factors produced by osteoblasts grown on microstructured Ti were sufficient to induce co-cultured MSC differentiation to osteoblasts. Silencing studies showed that this was due to signaling via alpha2beta1 integrins in osteoblasts on the substrate surface and paracrine action of secreted Dkk2. Thus, human MSCs are sensitive to substrate properties that induce osteoblastic differentiation; osteoblasts interact with these surface properties via alpha2beta1 and secrete Dkk2, which acts on distal MSCs.

The role of phospholipase D in osteoblast response to titanium surface microstructure.

Biomaterial surface properties such as microtopography and energy can change cellular responses at the cell-implant interface. Phospholipase D (PLD) is required for the differentiation of osteoblast-like MG63 cells on machined and grit-blasted titanium surfaces. Here, we determined if PLD is also required on microstructured/high-energy substrates and the mechanism involved. shRNAs for human PLD1 and PLD2 were used to silence MG63 cells. Wild-type and PLD1 or PLD1/2 silenced cells were cultured on smooth-pretreatment surfaces (PT); grit-blasted, acid-etched surfaces (SLA); and SLA surfaces modified to have higher surface energy (modSLA). PLD was inhibited with ethanol or activated with 24,25-dihydroxyvitamin-D(3) [24R,25(OH)(2)D(3)]. As surface roughness/energy increased, PLD mRNA and activity increased, cell number decreased, osteocalcin and osteoprotegerin increased, and protein kinase C (PKC) and alkaline phosphatase specific activities increased. Ethanol inhibited PLD and reduced surface effects on these parameters. There was no effect on these parameters after knockdown of PLD1, but PLD1/2 double knockdown had effects comparableto PLD inhibition. 24R,25(OH)(2)D(3) increased PLD activity and the production of osteocalcin and osteoprotegerin, but decreased cell number on the rough/high-energy surfaces. These results confirm that surface roughness/energy-induced PLD activity is required for osteoblast differentiation and that PLD2 is the main isoform involved in this pathway. PLD is activated by 24R,25(OH)(2)D(3) in a surface-dependent manner and inhibition of PLD reduces the effects of surface microstructure/energy on PKC, suggesting that PLD mediates the stimulatory effect of microstructured/high-energy surfaces via PKC-dependent signaling.

The influence of surface energy on early adherent events of osteoblast on titanium substrates.

Surface energy of implant material is one of the important factors in the process of osseointegration. How surface energy regulates the signaling pathway of osteoblasts, however, is not well understood. Cell adhesion is one of the first steps essential to subsequent proliferation and differentiation of bone cells before tissue formation. Our present study was designed to investigate how surface energy may influence the early adhesion of human alveolar osteoblasts (AOBs). Substrates applied were two groups of titanium disks: (1) hydrophobic sandblasted and acid-etched (SLA) surfaces; (2) chemically modified hydrophilic SLA (modSLA) ones. Cell morphology and cell attachment were examined by scanning electron microscopy (SEM). Defined cytoskeletal actin organization was immunohistochemically examined using confocal laser scanning microscopy. RT-PCR was applied to detect and to compare the expression of focal adhesion kinase (FAK) of osteoblasts cultured on the two groups of substrates. The attachment rates of AOBs cultured on modSLA substrates were significantly higher than the cells on SLA ones within 3 h. AOBs on modSLA developed more defined actin stress fibers after 6 h of attachment. FAK expression was comparably higher on modSLA after 6 h. Within the limitation of the current study, higher surface energy of titanium surfaces enhanced the cell adhesion in the early stage of cell response and may work through influencing the expression of adhesion-associated molecules.

Proliferation, behavior, and cytokine gene expression of human umbilical vascular endothelial cells in response to different titanium surfaces.

Success of dental implantation is initially affected by wound healing of both, hard and soft tissues. Endothelial cells (ECs) are involved as crucial cells in the angiogenesis and inflammation process of wound healing. In the present study, proliferation, mobility, cluster formation, and gene expression of angiogenesis-related molecules of human umbilical vascular endothelial cells (HUVECs) were investigated on titanium surfaces with different roughnesses: acid-etched (A), coarse-grit-blasted and acid-etched (SLA) surfaces, as well as on hydrophilic modified modA and modSLA surfaces. Cell behaviors were analyzed by proliferation assay and time-lapse microscopy, gene expression was analyzed by real time PCR. Results showed that cell proliferation, mobility, and cluster formation were highest on modA surfaces compared with all other surfaces. HUVECs moved slowly and exhibited seldom cell aggregation on SLA and modSLA surfaces during the whole observing period of 120 h. The gene expressions of the angiogenesis-related factors von Willebrand factor, thrombomodulin, endothelial cell protein C receptor, and adhesion molecules intercellular adhesion molecule-1 and E-selectin were most enhanced on modSLA surfaces. These results suggest that modA surface is optimal for proliferation and angiogenic behavior of ECs. However, modSLA surface seems to promote ECs to express angiogenesis-related factor genes, which play essential roles in controlling inflammation and revascularization of wound healing.

The roles of Wnt signaling modulators Dickkopf-1 (Dkk1) and Dickkopf-2 (Dkk2) and cell maturation state in osteogenesis on microstructured titanium surfaces.

Osteoblast differentiation on tissue culture polystyrene (TCPS) requires Wnt/beta-catenin signaling, regulating modulators of the Wnt pathway like Dickkopf-1 (Dkk1) and Dkk2. Osteoblast differentiation is increased on microstructured titanium (Ti) surfaces compared to TCPS; therefore, we hypothesized that surface topography and hydrophilicity affect Dkk1 and Dkk2 expression and that their roles in osteoblast differentiation on Ti differs depending on cell maturation state. Human osteoblast-like MG63 cells, normal human osteoblasts (HOBs), and human mesenchymal stem cells (MSCs), as well as MG63 cells stably silenced for Dkk1 or Dkk2 were grown for 6 days on TCPS and Ti surfaces (PT [Ra<0.2 microm], SLA [Ra=4 microm], modSLA [hydrophilic-SLA]). Dkk1 and Dkk2 mRNA and protein increased on SLA and modSLA for all cell types, but exogenous rhDkk1 and rhDkk2 affected MSCs differently than MG63 cells and HOBs. Silencing Dkk1 reduced MG63 cell number on TCPS and PT, but increased differentiation on these substrates. Silencing Dkk2 reduced stimulatory effects of SLA and modSLA on osteoblast differentiation; Dkk2 but not Dkk1 restored these effects. Antibodies to Dkk1 or Dkk2 specifically blocked substrate-dependent changes caused by the proteins, demonstrating their autocrine action. This indicates major roles for Dkk1 and the canonical Wnt pathway in early-stage differentiation, and for Dkk2 and Wnt/Ca2+-dependent signaling in late-stage differentiation on microstructured and hydrophilic surfaces, during osseointegration.

A comparison study of the osseointegration of zirconia and titanium dental implants. A biomechanical evaluation in the maxilla of pigs.

OBJECTIVES: The purpose of the present study was to investigate the osseointegration of microstructured zirconia implants in comparison with sandblasted and acid-etched (SLA) titanium implants in a biomechanical study. MATERIALS: Zirconia implants (4.1 mm in diameter, 10 mm in length) were produced using a new low pressure injection molding technique. After that the implants were acid-etched with hydrofluoric acid. Standard Ti-SLA implants of the exact same shape served as controls. Six months after extraction of incisors 2 and 3, 16 adult pigs received a total of 64 implants in the maxillae. After 4, 8, and 12 weeks the animals were sacrificed, and 59 implants could be analyzed to removal torque (RTQ) testing. RESULTS: The mean RTQ values for zirconia implants were 42.4 Ncm at 4 weeks, 69.6 Ncm at 8 weeks, and 69.3 Ncm at 12 weeks of healing, whereas RTQ values for the Ti-SLA implants were 42.1 Ncm, 75.0 Ncm, and 73.1 Ncm at corresponding time intervals. There is no statistical difference in RTQ values between Ti-SLA and zirconia implants at 8 weeks. CONCLUSIONS: Within the limits of the present study it was concluded that acid-etching of zirconia implants enhances bone apposition resulting in RTQ values which were equivalent to that of Ti-SLA.

The challenges in artificial muscle research to treat incontinence.

Sphincters to guarantee continence are in principal the simplest muscles, because only two states (closed and open) seem to be important. The healthy urinary sphincter, however, provides dynamic components. During the filling phase the increase in tonus prevents urinary loss. The sphincter rapidly responds to pressure pulses caused, for example, by coughing. Contemporary artificial sphincters, however, merely generate two states and often induce atrophy and erosion. Hence the success of commercially available, continually improved implants is still limited. This communication reviews two physical principles, shape memory alloys and electrically activated polymer nanostructures, for applications in artificial sphincters which adapt the pressure acting on the urethra and react to stress situations such as coughing. The application of these principles allows intermittent reduction of pressure on the urethra, thus involving significantly less atrophy. The fabrication of reliably working nanostructures, however, is ambitious and will need time-consuming, high-level engineering.