Cellular responses to 3D printed dental resins produced using a manufacturer recommended printer versus a third party printer.

PURPOSE: The aim of this study was to evaluate the influence of different 3D dental resins, using a manufacturer recommended printer and a third-party printer, on cellular responses of human gingival cells. MATERIALS AND METHODS: Three NextDent resins (Denture 3D+, C&B MFH and Crowntec) were used to produce specimens on printers NextDent 5100 (groups ND, NC and NT, respectively) and Phrozen Sonic Mini 4K (groups PD, PC and PT, respectively). Human gingival fibroblasts were cultured and biocompatibility was evaluated on days 1, 3 and 7. IL-6 and IL-8 concentrations were evaluated at 3 days using ELISA. Surface roughness was evaluated by a contact profilometer. SEM and fluorescence micrographs were analyzed at days 1 and 7. Statistical analyses were performed using SPSS and mean differences were tested using ANOVA and post-hoc Tukey tests (P < .05). RESULTS: There was an increase in cellular viability after 7 days in groups PC and PT, when compared to group PD. ND group resulted in higher concentration of IL-6 when compared to PT group. SEM and fluorescence micrographs showed less adhesion and thinner morphology of fibroblasts from group PD. No significant differences were found regarding surface roughness. CONCLUSION: The use of different printers or resins did not seem to influence surface roughness. NextDent 5100 and Phrozen Sonic Mini 4K produced resins with similar cellular responses in human gingival fibroblasts. However, Denture 3D+ resin resulted in significantly lower biocompatibility, when compared to C&B MFH and Crowntec resins. Further testing is required to support its long-term use, required for complete dentures.

Cell Biological and Antibacterial Evaluation of a New Approach to Zirconia Implant Surfaces Modified with MTA.

Peri-implantitis continues to be one of the major reasons for implant failure. We propose a new approach to the incorporation of MTA into zirconia implant surfaces with Nd:YAG laser and investigate the biological and the microbiological responses of peri-implant cells. Discs of zirconia stabilized with yttria and titanium were produced according to the following four study groups: Nd:YAG laser-textured zirconia coated with MTA (Zr MTA), Nd:YAG laser-textured zirconia (Zr textured), polished zirconia discs, and polished titanium discs (Zr and Ti). Surface roughness was evaluated by contact profilometry. Human osteoblasts (hFOB), gingival fibroblasts (HGF hTERT) and S. oralis were cultured on discs. Cell adhesion and morphology, cell differentiation markers and bacterial growth were evaluated. Zr textured roughness was significantly higher than all other groups. SEM images reveal cellular adhesion at 1 day in all samples in both cell lines. Osteoblasts viability was lower in the Zr MTA group, unlike fibroblasts viability, which was shown to be higher in the Zr MTA group compared with the Zr textured group at 3 and 7 days. Osteocalcin and IL-8 secretion by osteoblasts were higher in Zr MTA. The Zr textured group showed higher IL-8 values released by fibroblasts. No differences in S. oralis CFUs were observed between groups. The present study suggests that zirconia implant surfaces coated with MTA induced fibroblast proliferation and osteoblast differentiation; however, they did not present antibacterial properties.

Bio-Piezoelectric Ceramic Composites for Electroactive Implants-Biological Performance.

Barium titanate (BaTiO3) piezoelectric ceramic may be a potential alternative for promoting osseointegration due to its piezoelectric properties similar to bone electric potentials generated in loading function. In this sense, the aim of this in vitro study was to evaluate the cellular response of human osteoblasts and gingival fibroblasts as well as the impact on S. oralis when in contact with BaTiO3 functionalized zirconia implant surfaces with piezoelectric properties. Zirconia discs with BaTiO3 were produced and contact poling (piezo activation) was performed. Osteoblasts (hFOB 1.19), fibroblasts (HGF hTERT) and S. oralis were culture on discs. Cell viability and morphology, cell differentiation markers, bacterial adhesion and growth were evaluated. The present study suggests that zirconia composite surfaces with the addition of piezoelectric BaTiO3 are not cytotoxic to peri-implant cells. Also, they seem to promote a faster initial osteoblast differentiation. Moreover, these surfaces may inhibit the growth of S. oralis by acting as a bacteriostatic agent over time. Although the piezoelectric properties do not affect the cellular inflammatory profile, they appear to enable the initial adhesion of bacteria, however this is not significant over the entire testing period. Furthermore, the addition of non-poled BaTiO3 to zirconia may have a potential reduction effect on IL-6 mediated-inflammatory activity in fibroblasts.

Modification of Zirconia Implant Surfaces by Nd:YAG Laser Grooves: Does It Change Cell Behavior?

The aim of this study was to evaluate gingival fibroblasts and human osteoblasts' response to textured Nd:YAG laser microgrooves, with different dimensions, on zirconia implant surfaces. A total of 60 zirconia disks (8 mm in diameter and 2 mm in thickness) were produced and divided between four study groups (N = 15): three laser-textured (widths between 125.07 ± 5.29 µm and 45.36 ± 2.37 µm and depth values from 50.54 ± 2.48 µm to 23.01 ± 3.79 µm) and a control group without laser treatment. Human osteoblasts and gingival fibroblasts were cultured on these surfaces for 14 days. FEG-SEM (Field Emission Gun-Scanning Electron Microscope) images showed cellular adhesion at 24 h, with comparable morphology in all samples for both cell types. A similar cell spreading within the grooves and in the space between them was observed. Cell viability increased over time in all study groups; however, no differences were found between them. Additionally, proliferation, ALP (Alkaline phosphatase) activity, collagen type I, osteopontin and interleukin levels were not significantly different between any of the study groups for any of the cell types. Analysis of variance to compare parameters effect did not reveal statistically significant differences when comparing all groups in the different tests performed. The results obtained revealed similar cell behavior based on cell viability and differentiation on different microtopographic laser grooves, compared to a microtopography only established by sandblasting and acid-etching protocol, the reference surface treatment on zirconia dental implants.

Biomimetic Implant Surfaces and Their Role in Biological Integration-A Concise Review.

BACKGROUND: The increased use of dental implants in oral rehabilitation has been followed by the development of new biomaterials as well as improvements in the performance of biomaterials already in use. This triggers the need for appropriate analytical approaches to assess the biological and, ultimately, clinical benefits of these approaches. AIMS: To address the role of physical, chemical, mechanical, and biological characteristics in order to determine the critical parameters to improve biological responses and the long-term effectiveness of dental implant surfaces. DATA SOURCES AND METHODS: Web of Science, MEDLINE and Lilacs databases were searched for the last 30 years in English, Spanish and Portuguese idioms. RESULTS: Chemical composition, wettability, roughness, and topography of dental implant surfaces have all been linked to biological regulation in cell interactions, osseointegration, bone tissue and peri-implant mucosa preservation. CONCLUSION: Techniques involving subtractive and additive methods, especially those involving laser treatment or embedding of bioactive nanoparticles, have demonstrated promising results. However, the literature is heterogeneous regarding study design and methodology, which limits comparisons between studies and the definition of the critical determinants of optimal cell response.

Human Gingival Fibroblast and Osteoblast Behavior on Groove-Milled Zirconia Implant Surfaces.

Two type of cells representing periodontal hard tissues (osteoblasts) and soft tissues (fibroblasts) were evaluated in response to microgroove-milled zirconia surfaces. A total of 90 zirconia discs were randomly assigned to four width-standardized milling microgroove-textured groups and a control group without grooves (UT). The sandblast and acid-etch protocol were applied to all samples. Both cell lines were cultured on zirconia discs from 1 day up to 14 days. Cell morphology and adhesion were evaluated after 1 day of culturing. Cell viability and proliferation of the cells were measured. Alkaline phosphatase activity, collagen I, osteopontin, interleukin 1ß and interleukin 8 secretions were assessed at predefined times. The results obtained were presented in the form of bar graphs as means and standard deviations. Multi comparisons between groups were evaluated using two-away ANOVA or Mann−Whitney tests, and a p-value < 0.05 was established. Group comparisons with regard to cell viability, proliferation and secretion of collagen I, interleukin-1ß and interleukin 8 revealed no statistically significant differences. The alkaline phosphatase activity and osteopontin secretion were significantly higher in the group with a large groove compared to the small one and the control group. Nevertheless, the viability of gingival and bone cells did not appear to be affected by the milled microgroove texture compared to the conventional sandblasted and acid-etched texture, but they seem to influence osteoblasts' cellular differentiation.

Peri-implant cell response on groove and pore-textured zirconia surfaces.

OBJECTIVES: This study aimed to assess the independent influence of grooves and pores texturized by milling on gold-standard zirconia implant surfaces. METHODS: Milled groove and pore textured with equivalent width, depth, and spacing on zirconia discs were produced using press and sintering techniques. All samples were sandblasted and acid-etched (SBAE), and untextured discs were used as controls. Osteoblasts and fibroblasts were cultured on discs for 14 days. Field emission gun-scanning electron microscopy (FEG-SEM) was used to observe cellular adhesion and morphology. Cell viability and proliferation assays were performed. Additionally, alkaline phosphatase activity, collagen type I, and osteopontin were evaluated at pre-defined time points. Results are presented as mean and standard deviation (SD), group comparisons were tested using one-way ANOVA (Tukey's post-hoc), and significance was set at P < 0.05. RESULTS: FEG-SEM images revealed cellular adhesion at 24 h in all samples with differences in distribution. Although both cell lines showed increased cell viability and differentiation cell markers such as collagen and osteopontin over time, statistically significant differences between groups were found in none of the quantitative study variables (P > 0.05). CONCLUSION: The results suggest similar cellular behavior between different patterns with similar dimensions and between them and microtopography by SBAE protocol currently used as the gold-standard for zirconia dental implants. The addition of pore and groove microtextures to the gold-standard zirconia dental implant surfaces treated with SBAE does not seem to be an asset in the cellular behavior of the hard and soft tissue cells.

Laser surface treatment on Yttria-stabilized zirconia dental implants: Influence on cell behavior.

Yttria-stabilized zirconia (YSZ) is being proposed as an alternative material to Titanium for dental implants due to its aesthetic and biocompatibility properties. However, is it yet to define the optimal surface treatment to improve YSZ bioactivy. Texturization is a promising approach, but the biological role of patterned YSZ surfaces in cell cultures is yet to be determined. Thus, cellular behavior of osteoblasts and fibroblasts in contact with groove-texturized YSZ surfaces was investigated. YSZ discs were groove-textured by conventional milling and Nd:YAG laser. All samples including control were sandblasted and acid-etched. Human osteoblasts and fibroblasts were cultured on discs for 14 days. Morphology and cellular adhesion were observed. Cell viability, interleukin-1ß, osteopontin, collagen type I prodution, alkaline phosphatase activity, and interleukin-8 were measured. YSZ texturization by conventional milling improved osteoblasts viability and differentiation when compared to laser texturization. Fibroblasts behavior did not seem to be influenced by the texturing technique. Compared to sandblasting and acid etching currently used as gold standard for zirconia dental implants no superiority of macrotexturization was found.

Bioactive-Enhanced Polyetheretherketone Dental Implant Materials: Mechanical Characterization and Cellular Responses.

The aim of this study was to characterize the mechanical properties of a bioactive-modified polyetheretherketone (PEEK) manufacturing approach for dental implants and to compare the in vitro biological behavior with titanium alloy (Ti6Al4V) as the reference. PEEK, PEEK with 5% hydroxyapatite (HA), PEEK with 5% beta-tricalcium phosphate (ßTCP), and Ti6Al4V discs were produced using hot pressing technology to create a functionally graded material (FGM). Surface roughness values (Ra, Rz), water contact angle, shear bond strength, and Vickers hardness tests were performed. Human osteoblasts and gingival fibroblasts bioactivity was evaluated by a resazurin-based method, alkaline phosphatase activity (ALP), and confocal laser scanning microscopy (CLSM) images of fluorescent-stained fibroblasts. Morphology and cellular adhesion were confirmed using field emission gun-scanning electron microscopy (FEG-SEM). Group comparisons were tested using analysis of variance (Tukey post hoc test), α = .05. All groups presented similar roughness values (P > .05). Ti6Al4V group was found to have the highest contact angle (P < .05). Shear bond strength and Vickers hardness of different PEEK materials were similar (P > .05); however, the mean values in the Ti6Al4V group were significantly higher when compared with those of the other groups (P < .05). Cell viability and proliferation of osteoblast and fibroblast cells were higher in the PEEK group (P < .05). PEEK-ßTCP showed the highest significant ALP activity over time (P < .05 at 14 days of culture). An enhanced bone and soft-tissue cell behavior on pure PEEK was obtained to the gold standard (Ti6Al4V) with equivalent roughness. The results substantiate the potential role of chemical composition rather than physical properties of materials in biological responses. The addition of 5% HA or ßTCP by FGM did not enhance PEEK mechanical properties or periodontal cell behavior.

Evaluation of Saliva Stability for NMR Metabolomics: Collection and Handling Protocols.

Maintaining a salivary metabolic profile upon sample collection and preparation is determinant in metabolomics. Nuclear magnetic resonance (NMR) spectroscopy was used to identify metabolite changes during short-term storage, at room temperature (RT)/4 °C/-20 °C, and after sample preparation, at RT/4 °C (mimicking typical clinical/laboratory settings). Interestingly, significant metabolic inter-individual and inter-day variability were noted, probably determining sample stability to some extent. After collection, no changes were noted at -20 °C (at least for 4 weeks). RT storage induced decreases in methylated macromolecules (6 h); lactate (8 h); alanine (12 h); galactose, hypoxanthine, pyruvate (24 h); sarcosine, betaine, choline, N-acetyl-glycoproteins (48 h), while acetate increased (48 h). Less, but different, changes were observed at 4 °C, suggesting different oral and microbial status at different temperatures (with a possible contribution from inter-individual and inter-day variability), and identifying galactose, hypoxanthine, and possibly, choline esters, as potential general stability indicators. After preparation, addition of NaN3 did not impact significantly on saliva stabilization, neither at RT nor at 4 °C, although its absence was accompanied by slight increases in fucose (6.5 h) and proline (8 h) at RT, and in xylose (24 h) at 4 °C. The putative metabolic origins of the above variations are discussed, with basis on the salivary microbiome. In summary, after collection, saliva can be stored at RT/4 °C for up to 6 h and at -20 °C for at least 4 weeks. Upon preparation for NMR analysis, samples are highly stable at 25 °C up to 8 h and at 4 °C up to 48 h, with NaN3 addition preventing possible early changes in fucose, proline (6-8 h), and xylose (24 h) levels.

Gingival fibroblasts behavior on bioactive zirconia and titanium dental implant surfaces produced by a functionally graded technique.

Adding a biological apatite layer to the implant surface enhances bone healing around the implant. Objective This study aimed to characterize the mechanical properties and test human gingival fibroblasts behavior in contact with Zirconia and Titanium bioactive-modified implant materials. Methodology 6 groups were considered: Titanium (Ti6Al4V), Ti6Al4V with 5% HA and 5% ßTCP, Zirconia (YTZP), YTZP with 5% HA and 5% ßTCP. For each group, we produced discs using a novel fabrication method for functionally graded materials, under adequate conditions for etching and grit-blasting to achieve equivalent surface microroughness among the samples. Surface roughness (Ra, Rz), water contact angle, shear bond strength, and Vickers hardness were performed. Human gingival fibroblasts immortalized by hTERT gene from the fourth passage, were seeded on discs for 14 days. Cell viability and proliferation were assessed using a resazurin-based method, and cellular adhesion and morphology using field emission gun scanning electron microscopy (FEG-SEM). After 3 days of culture, images of fluorescent nucleic acid stain were collected by confocal laser scanning microscopy (CLSM). Results Results were presented as mean ± standard deviation (SD). We compared groups using one-way ANOVA with Tukey's post-hoc test, and significance level was set at p<0.05. After 14 days of culture, cell viability and proliferation were significantly higher in YTZP group than in other groups (p<0.05). Samples of YTZP-ßTCP presented significantly higher wettability (p<0.05); yet, we observed no improvement in cell behavior on this group. Fibroblast spreading and surface density were more evident on YTZP specimens. Adding calcium-phosphate bioactive did not alter the tested mechanical properties; however, Ti6Al4V material shear bond strength was statistically higher than other groups (p<0.05). Conclusion Adding bioactive materials did not improve soft-tissue cell behavior. When compared to other zirconia and titanium groups, pure zirconia surface improved adhesion, viability and proliferation of fibroblasts. Cell behavior seems to depend on surface chemical composition rather than on surface roughness.

Laser Nd:YAG patterning enhance human osteoblast behavior on zirconia implants.

Zirconia has been regarded as a promising material for dental implants, and Nd:YAG laser treatment has been proposed as a potential strategy to improve its bioactivity. The main aim of the present study was to evaluate the in vitro behavior of human fetal osteoblasts in contact with laser-textured zirconia implant surfaces assessing the effect of different texture patterns, spacing between laser passes and number of laser passes. Zirconia discs were produced and treated with Nd:YAG laser according to test group variables: texture (microgrooves and micropillar array), distance between surface features (25 µm, 30 µm and 35 µm), and laser passes [1, 2, 4, and 8]. Untextured sandblasted and acid-etched zirconia discs (SBAE) were used as controls. Human osteoblasts (hFOB 1.19) were cultured for 14 days on test and control samples. Morphology and cellular adhesion were observed using scanning electron microscopy (SEM). Cell viability and proliferation were evaluated at 1, 3, 7, and 14 days using a commercial resazurin-based method. Collagen type I was evaluated at 3 days using ELISA. Alkaline phosphatase (ALP) activity was evaluated at 7 days using a colorimetric enzymatic technique. Group comparisons were tested using ANOVA or Mann-Whitney test (Tukey's post hoc) using statistical software, and significance was set at p < 0.05. Cell viability and proliferation increased over time for all groups with statistically higher values for laser-textured groups when compared with control at 7 and 14 days in culture (p < 0.05). Collagen type I levels were higher for study groups (p < 0.05) when compared with control group. No statistically differences were detected for ALP activity levels between texture and control groups (p > 0.05). The results suggest that laser-machined zirconia implant surfaces may benefit biological osteoblast response. However, the type of texture, spacing at the range of 25-35 µm, and number of laser passes did not seem to be relevant variables.

Gingival fibroblasts behavior on bioactive zirconia and titanium dental implant surfaces produced by a functionally graded technique

Abstract Adding a biological apatite layer to the implant surface enhances bone healing around the implant. Objective This study aimed to characterize the mechanical properties and test human gingival fibroblasts behavior in contact with Zirconia and Titanium bioactive-modified implant materials. Methodology 6 groups were considered: Titanium (Ti6Al4V), Ti6Al4V with 5% HA and 5% ßTCP, Zirconia (YTZP), YTZP with 5% HA and 5% ßTCP. For each group, we produced discs using a novel fabrication method for functionally graded materials, under adequate conditions for etching and grit-blasting to achieve equivalent surface microroughness among the samples. Surface roughness (Ra, Rz), water contact angle, shear bond strength, and Vickers hardness were performed. Human gingival fibroblasts immortalized by hTERT gene from the fourth passage, were seeded on discs for 14 days. Cell viability and proliferation were assessed using a resazurin-based method, and cellular adhesion and morphology using field emission gun scanning electron microscopy (FEG-SEM). After 3 days of culture, images of fluorescent nucleic acid stain were collected by confocal laser scanning microscopy (CLSM). Results Results were presented as mean ± standard deviation (SD). We compared groups using one-way ANOVA with Tukey's post-hoc test, and significance level was set at p<0.05. After 14 days of culture, cell viability and proliferation were significantly higher in YTZP group than in other groups (p<0.05). Samples of YTZP-ßTCP presented significantly higher wettability (p<0.05); yet, we observed no improvement in cell behavior on this group. Fibroblast spreading and surface density were more evident on YTZP specimens. Adding calcium-phosphate bioactive did not alter the tested mechanical properties; however, Ti6Al4V material shear bond strength was statistically higher than other groups (p<0.05). Conclusion Adding bioactive materials did not improve soft-tissue cell behavior. When compared to other zirconia and titanium groups, pure zirconia surface improved adhesion, viability and proliferation of fibroblasts. Cell behavior seems to depend on surface chemical composition rather than on surface roughness.

Hard and Soft Tissue Cell Behavior on Polyetheretherketone, Zirconia, and Titanium Implant Materials

PURPOSE: The aim of this study was to characterize and compare the behavior of human osteoblasts and human gingival fibroblasts in contact with polyetheretherketone (PEEK), zirconia, and titanium implant surface materials. MATERIALS AND METHODS: PEEK, yttria-stabilized zirconia (YTZP), and titanium discs were produced under appropriate and similar conditions to achieve controlled surface features. Human osteoblasts and human gingival fibroblasts were cultured on discs for 14 days. Cell viability and proliferation were evaluated using a resazurin-based method. Morphology and cellular adhesion were observed using field emission gun-scanning electron microscopy (FEG-SEM). Alkaline phosphatase (ALP) activity and bone cell mineralization were evaluated on osteoblasts. Confocal laser scanning microscopy (CLSM) images of fluorescent-stained fibroblasts were obtained at 7 and 14 days of the culture. Results were presented as mean and standard deviation (SD). Group comparisons were tested using analysis of variance (ANOVA) (Tukey's post hoc) with appropriate statistical software, and significance was set at P < .05. RESULTS: Cell viability and proliferation were higher in PEEK and YTZP groups compared with titanium on osteoblast cells (P < .05, all time points) and on fibroblasts (P < .05, 7 and 14 days). All groups showed an increase in ALP activity over time, which was not significant. Mineralization patterns demonstrated an increase in mineral content over time, which was more apparent in the YTZP group. Cell spreading was more evident on PEEK and YTZP specimens. CONCLUSION: The results suggest increased adhesion, viability, and proliferation of osteoblasts and gingival fibroblasts on zirconia and PEEK surfaces compared with titanium. These results are correlated with the increased wettability of these materials.