Gingival fibroblast response to (hybrid) ceramic implant reconstruction surfaces is modulated by biomaterial type and surface treatment.

OBJECTIVES: Surface characteristics of implant reconstructions determine the gingival fibroblast (GF) response and thus soft tissue integration (STI). However, for monolithic implant reconstructions it is unknown whether the (hybrid) ceramic biomaterial type and its surface treatment affect GF response. Therefore, this investigation examined the influence of the implant reconstruction biomaterials hybrid ceramic (HC), lithium disilicate ceramic (LS), 4 and 5 mol% yttria partially stabilized zirconiumdioxide ceramics (4/5Y-PSZ) and their surface treatment - machining, polishing or glazing - on surface characteristics and GF response. METHODS: After characterization of surface topography and wettability by scanning electron microscopy, interferometry and contact angle measurement, the adhesion, morphology, metabolic activity and proliferation of GFs from six donors was investigated by fluorescent staining and a resazurin-based assay at days 1, 3 and 7. Titanium (Ti) served as control. RESULTS: Biomaterial type and surface treatment affected the GF response in a topography-dependent manner. Smooth polished and glazed surfaces demonstrated enhanced GF adhesion and earlier proliferation onset compared to rough machined surfaces. Due to minor differences in surface topography of polished and glazed surfaces, however, the GF response was similar for polished and glazed HC, LS, 4- and 5Y-PSZ as well as Ti. SIGNIFICANCE: Within the limits of the present investigation, polishing and glazing of machined HC, LS and 4/5Y-PSZ can be recommended to support STI-relevant cell functions in GF. Since the GF response on polished and glazed HC, LS, 4- and 5Y-PSZ surfaces and the Ti control was comparable, this investigation proofed equal cytocompatibility of these surfaces in vitro.

Analysis of soft tissue integration-supportive cell functions in gingival fibroblasts cultured on 3D printed biomaterials for oral implant-supported prostheses.

To date, it is unknown whether 3D printed fixed oral implant-supported prostheses can achieve comparable soft tissue integration (STI) to clinically established subtractively manufactured counterparts. STI is mediated among others by gingival fibroblasts (GFs) and is modulated by biomaterial surface characteristics. Therefore, the aim of the present work was to investigate the GF response of a 3D printed methacrylate photopolymer and a hybrid ceramic-filled methacrylate photopolymer for fixed implant-supported prostheses in the sense of supporting an STI. Subtractively manufactured samples made from methacrylate polymer and hybrid ceramic were evaluated for comparison and samples from yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP), comprising well documented biocompatibility, served as control. Surface topography was analyzed by scanning electron microscopy and interferometry, elemental composition by energy-dispersive x-ray spectroscopy, and wettability by contact angle measurement. The response of GFs obtained from five donors was examined in terms of membrane integrity, adhesion, morphogenesis, metabolic activity, and proliferation behavior by a lactate-dehydrogenase assay, fluorescent staining, a resazurin-based assay, and DNA quantification. The results revealed all surfaces were smooth and hydrophilic. GF adhesion, metabolic activity and proliferation were impaired by 3D printed biomaterials compared to subtractively manufactured comparison surfaces and the 3Y-TZP control, whereas membrane integrity was comparable. Within the limits of the present investigation, it was concluded that subtractively manufactured surfaces are superior compared to 3D printed surfaces to support STI. For the development of biologically optimized 3D printable biomaterials, consecutive studies will focus on the improvement of cytocompatibility and the synthesis of STI-relevant extracellular matrix constituents.

Identification of Zirconia Particle Uptake in Human Osteoblasts by ToF-SIMS Analysis and Particle-Size Effects on Cell Metabolism.

As the use of zirconia-based nano-ceramics is rising in dentistry, the examination of possible biological effects caused by released nanoparticles on oral target tissues, such as bone, is gaining importance. The aim of this investigation was to identify a possible internalization of differently sized zirconia nanoparticles (ZrNP) into human osteoblasts applying Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS), and to examine whether ZrNP exposure affected the metabolic activity of the cells. Since ToF-SIMS has a low probing depth (about 5 nm), visualizing the ZrNP required the controlled erosion of the sample by oxygen bombardment. This procedure removed organic matter, uncovering the internalized ZrNP and leaving the hard particles practically unaffected. It was demonstrated that osteoblasts internalized ZrNP within 24 h in a size-dependent manner. Regarding the cellular metabolic activity, metabolization of alamarBlue by osteoblasts revealed a size- and time-dependent unfavorable effect of ZrNP, with the smallest ZrNP exerting the most pronounced effect. These findings point to different uptake efficiencies of the differently sized ZrNP by human osteoblasts. Furthermore, it was proven that ToF-SIMS is a powerful technique for the detection of zirconia-based nano/microparticles that can be applied for the cell-based validation of clinically relevant materials at the nano/micro scale.

Zirconia fixed dental prostheses fabricated by 3D gel deposition show higher fracture strength than conventionally milled counterparts.

Zirconia restorations, which are fabricated by additive 3D gel deposition and do not require glazing like conventional restorations, were introduced as "self-glazed" zirconia restorations into dentistry. This in vitro investigation characterized the surface layer, microstructure and the fracture and aging behavior of "self-glazed" zirconia (Y-TZPSG) three-unit fixed dental prostheses (FDP) and compared them to conventionally CAD/CAM milled and glazed controls (Y-TZPC-FDPs). For this purpose, the FDPs were analyzed by (focused ion beam) scanning electron microscopy, laserscanning microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction and a dynamic and static loading test. For the latter, half of the samples of each material group (n = 16) was subjected to 5 million cycles of thermocyclic loading (98N) in an aqueous environment in a chewing simulator. Afterwards, all FDPs were loaded to fracture. Y-TZPSG-FDPs demonstrated a comparable elemental composition but higher surface microstructural homogeneity and fracture strength compared to Y-TZPC-FDPs. Microstructural flaws within the FDPs' surfaces were identified as fracture origins. The high fracture strength of the Y-TZPSG-FDPs was attributed to a finer-grained microstructure with fewer surface flaws compared to the Y-TZPC-FDPs which showed numerous flaws in the glaze overlayer. A decrease in fracture strength after dynamic loading from 5165N to 4507N was observed for the Y-TZPSG-FDPs, however, fracture strength remained statistically significantly above the one measured for Y-TZPC-FDPs (before chewing simulation: 1923N; after: 2041N). Within the limits of this investigation, it can therefore be concluded that Y-TZPSG appears to be stable for clinical application suggesting further investigations to prove clinical applicability.

3D printed zirconia dental implants with integrated directional surface pores combine mechanical strength with favorable osteoblast response.

Dental implants need to combine mechanical strength with promoted osseointegration. Currently used subtractive manufacturing techniques require a multi-step process to obtain a rough surface topography that stimulates osseointegration. Advantageously, additive manufacturing (AM) enables direct implant shaping with unique geometries and surface topographies. In this study, zirconia implants with integrated lamellar surface topography were additively manufactured by nano-particle ink-jetting. The ISO-14801 fracture load of as-sintered implants (516±39 N) resisted fatigue in 5-55 °C water thermo-cycling (631±134 N). Remarkably, simultaneous mechanical fatigue and hydrothermal aging at 90 °C significantly increased the implant strength to 909±280 N due to compressive stress generated at the seamless transition of the 30-40 µm thick, rough and porous surface layer to the dense implant core. This unique surface structure induced an elongated osteoblast morphology with uniform cell orientation and allowed for osteoblast proliferation, long-term attachment and matrix mineralization. In conclusion, the developed AM zirconia implants not only provided high long-term mechanical resistance thanks to the dense core along with compressive stress induced at the transition zone, but also generated a favorable osteoblast response owing to the integrated directional surface pores. STATEMENT OF SIGNIFICANCE: Zirconia ceramics are becoming the material of choice for metal-free dental implants, however significant efforts are required to obtain a rough/porous surface for enhanced osseointegration, along with the risk of surface delamination and/or microstructure variation. In this study, we addressed the challenge by additively manufacturing implants that seamlessly combine dense core with a porous surface layer. For the first time, a unique surface with a directional lamellar pore morphology was additively obtained. This AM implant also provided strength as strong as conventionally manufactured zirconia implants before and after long-term fatigue. Favorable osteoblast response was proved by in-vitro cell investigation. This work demonstrated the opportunity to AM fabricate novel ceramic implants that can simultaneously meet the mechanical and biological functionality requirements.

A systematic review and meta-analysis evaluating the survival, the failure, and the complication rates of veneered and monolithic all-ceramic implant-supported single crowns.

OBJECTIVE: To assess the survival, failure, and complication rates of veneered and monolithic all-ceramic implant-supported single crowns (SCs). METHODS: Literature search was conducted in Medline (PubMed), Embase, and Cochrane Central Register of Controlled Trials until September 2020 for randomized, prospective, and retrospective clinical trials with follow-up time of at least 1 year, evaluating the outcome of veneered and/or monolithic all-ceramic SCs supported by titanium dental implants. Survival and complication rates were analyzed using robust Poisson's regression models. RESULTS: Forty-nine RCTs and prospective studies reporting on 57 material cohorts were included. Meta-analysis of the included studies indicated an estimated 3-year survival rate of veneered-reinforced glass-ceramic implant-supported SCs of 97.6% (95% CI: 87.0%-99.6%). The estimated 3-year survival rates were 97.0% (95% CI: 94.0%-98.5%) for monolithic-reinforced glass-ceramic implant SCs, 96.9% (95% CI: 93.4%-98.6%) for veneered densely sintered alumina SCs, 96.3% (95% CI: 93.9%-97.7%) for veneered zirconia SCs, 96.1% (95% CI: 93.4%-97.8%) for monolithic zirconia SCs and only 36.3% (95% CI: 0.04%-87.7%) for resin-matrix-ceramic (RMC) SCs. With the exception of RMC SCs (p < 0.0001), the differences in survival rates between the materials did not reach statistical significance. Veneered SCs showed significantly (p = 0.017) higher annual ceramic chipping rates (1.65%) compared with monolithic SCs (0.39%). The location of the SCs, anterior vs. posterior, did not influence survival and chipping rates. CONCLUSIONS: With the exception of RMC SCs, veneered and monolithic implant-supported ceramic SCs showed favorable short-term survival and complication rates. Significantly higher rates for ceramic chipping, however, were reported for veneered compared with monolithic ceramic SCs.

Human osteoblast and fibroblast response to oral implant biomaterials functionalized with non-thermal oxygen plasma.

Plasma-treatment of oral implant biomaterials prior to clinical insertion is envisaged as a potential surface modification method for enhanced implant healing. To investigate a putative effect of plasma-functionalized implant biomaterials on oral tissue cells, this investigation examined the response of alveolar bone osteoblasts and gingival fibroblasts to clinically established zirconia- and titanium-based implant surfaces for bone and soft tissue integration. The biomaterials were either functionalized with oxygen-plasma in a plasma-cleaner or left untreated as controls, and were characterized in terms of topography and wettability. For the biological evaluation, the cell adhesion, morphogenesis, metabolic activity and proliferation were examined, since these parameters are closely interconnected during cell-biomaterial interaction. The results revealed that plasma-functionalization increased implant surface wettability. The magnitude of this effect thereby depended on surface topography parameters and initial wettability of the biomaterials. Concerning the cell response, plasma-functionalization of smooth surfaces affected initial fibroblast morphogenesis, whereas osteoblast morphology on rough surfaces was mainly influenced by topography. The plasma- and topography-induced differential cell morphologies were however not strong enough to trigger a change in proliferation behaviour. Hence, the results indicate that oxygen plasma-functionalization represents a possible cytocompatible implant surface modification method which can be applied for tailoring implant surface wettability.

Controlling osteoblast morphology and proliferation via surface micro-topographies of implant biomaterials.

Current research on surface modifications has yielded advanced implant biomaterials. Various implant surface modifications have been shown to be promising in improving bone target cell response, but more comprehensive studies whether certain implant surface modifications can directly target cell behavioural features such as morphogenesis and proliferation are needed. Here, we studied the response of primary alveolar bone cells on various implant surface modifications in terms of osteoblast morphology and proliferation in vitro. Analyses of surface modifications led to surface-related test parameters including the topographical parameters micro-roughness, texture aspect and surface enlargement as well as the physicochemical parameter surface wettability. We compared osteoblast morphology and proliferation towards the above-mentioned parameters and found that texture aspect and surface enlargement but not surface roughness or wettability exhibited significant impact on osteoblast morphology and proliferation. Detailed analysis revealed osteoblast proliferation as a function of cell morphology, substantiated by an osteoblast size- and morphology-dependent increase in mitotic activity. These findings show that implant surface topography controls cell behavioural morphology and subsequently cell proliferation, thereby opening the road for cell instructive biomaterials.

Prosthodontic Rehabilitation with Fixed Monolithic Translucent Zirconia Restorations: A Case History Report.

Recent attempts in the development of novel zirconia ceramics aim at improving its optical characteristics by increasing the yttria content to up to 5 mol% so that these ceramics can be used for the fabrication of stable and esthetic monolithic restorations. However, clinical evidence on the outcomes of such restorations is sparse. In this case report, monolithic inlays, partial crowns, tooth- and implant-supported single crowns, and fixed dental prostheses were fabricated out of a zirconia ceramic doped with 5 mol% yttria. The restorations in the present case history report showed a satisfying esthetic outcome and are in situ as inserted 18 months after insertion.

The clinical performance of all-ceramic implant-supported single crowns: A systematic review and meta-analysis.

OBJECTIVE: This review aimed at evaluating the survival and technical complication rates of all-ceramic implant-supported single crowns (SC). MATERIAL AND METHODS: Three electronic databases were searched for clinical studies conducted at ≥ 15 patients examining implant-supported all-ceramic SCs over ≥ 12 months. Survival rates of implants and restorations plus technical complication rates of SCs were calculated and tested for statistical correlation with confounding variables. Statistical analysis was performed using a negative binomial distribution model to calculate 5- and 10-year survival and complication estimates. RESULTS: Forty-one included studies reported on implant-supported SCs made of veneered and monolithic high-strength oxide ceramics, monolithic, and veneered glass-based ceramics and of a monolithic resin-nano-ceramic (RNC). Survival estimates for SCs of 93% (95% CI: 86.6%-96.4%) after 5 years and 94.4% (95% CI: 91.1%-96.5%) after 10 years were calculated, corresponding values for implant survival were 95.3% (95% CI: 90.6%-97.7%) and 96.2% (95% CI: 95.1%-97.1%). Technical complication rates after 5/10 years were as follows: chipping 9.0% (95% CI: 5.4%-14.8%)/2.7% (95% CI: 2.1%-3.5%), framework fractures 1.9% (95% CI: 0.7%-4.9%)/1.2% (95% CI: 1%-1.5%), screw loosening 3.6% (95% CI: 1.6%-8.4%)/5.2% (95% CI: 3.6%-7.5%), and decementations with 1.1% (95% CI: 0.4%-2.8%) after 5 years. Some confounding variables influenced the above-mentioned estimates significantly. CONCLUSIONS: All-ceramic implant-supported SCs showed-with the exception of a RNC material-high survival rates. However, failures and technical complications occurred which have to be considered when informing patients on the treatment with implant-supported all-ceramic SCs.

Clinical outcomes of partial and full-arch all-ceramic implant-supported fixed dental prostheses. A systematic review and meta-analysis.

OBJECTIVE: To assess the survival and technical complication rate of partial and full-arch all-ceramic implant-supported fixed dental prostheses (P-FDP/FA-FDP) and supporting implants. MATERIALS AND METHODS: An electronic search through three databases (MEDLINE/Pubmed, Cochrane Library, Embase) was conducted to identify relevant clinical studies with an observation period of at least 12 months, including ≥15 patients. Reconstruction and implant survival rates, technical complications and confounding variables such as processed/installed materials, retention mode and location in the mouth were obtained. Failure and complication rates were analyzed using standard Poisson regression models to calculate 5-year survival and complication estimates. RESULTS: A total of five studies for the P-FDP group and seven studies for the FA-FDP group were included, throughout evaluating veneered zirconia reconstructions. In the P-FDP group, reconstructions were located in posterior regions. Meta-analysis indicated survival estimates on the reconstruction level of 98.3% and 97.7% for P- and FA-FDPs after 5 years. However, chipping of the veneering ceramic was frequent, resulting in estimated 5-year complication rates of 22.8% (P-FDPs) and 34.8% (FA-FDPs). Five-year survival estimates of implants supporting P-FDPs and FA-FDPs of 98.5% and 99.4% were calculated, respectively. Including a total of 540 FDPs, one screw-loosening and 11 de-cementations were reported. Confounding variables were not found to have a significant influence on survival and complication rates. CONCLUSIONS: All-ceramic implant-supported P- and FA-FDPs comprising veneered zirconia frameworks showed high survival but clinically inacceptable fracture rates of the veneering ceramic. Their suitability with regard to this indication and a successful long-term outcome needs to be further evaluated.

Cellular transcriptional response to zirconia-based implant materials.

OBJECTIVE: To adequately address clinically important issues such as osseointegration and soft tissue integration, we screened for the direct biological cell response by culturing human osteoblasts and gingival fibroblasts on novel zirconia-based dental implant biomaterials and subjecting them to transcriptional analysis. METHODS: Biomaterials used for osteoblasts involved micro-roughened surfaces made of a new type of ceria-stabilized zirconia composite with two different topographies, zirconium dioxide, and yttria-stabilized zirconia (control). For fibroblasts smooth ceria- and yttria-stabilized zirconia surface were used. The expression of 90 issue-relevant genes was determined on mRNA transcription level by real-time PCR Array technology after growth periods of 1 and 7 days. RESULTS: Generally, modulation of gene transcription exhibited a dual dependence, first by time and second by the biomaterial, whereas biomaterial-triggered changes were predominantly caused by the biomaterials' chemistry rather than surface topography. Per se, modulated genes assigned to regenerative tissue processes such as fracture healing and wound healing and in detail included colony stimulating factors (CSF2 and CSF3), growth factors, which regulate bone matrix properties (e.g. BMP3 and TGFB1), osteogenic BMPs (BMP2/4/6/7) and transcription factors (RUNX2 and SP7), matrix collagens and osteocalcin, laminins as well as integrin ß1 and MMP-2. SIGNIFICANCE: With respect to the biomaterials under study, the screening showed that a new zirconia-based composite stabilized with ceria may be promising to provide clinically desired periodontal tissue integration. Moreover, by detecting biomarkers modulated in a time- and/or biomaterial-dependent manner, we identified candidate genes for the targeted analysis of cell-implant bioresponse during biomaterial research and development.