Mass Spectrometry-Based Proteomics of Poly(methylmethacrylate)-Embedded Bone.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a widely employed technique in proteomics research for studying the proteome biology of various clinical samples. Hard tissues, such as bone and teeth, are routinely preserved using synthetic poly(methyl methacrylate) (PMMA) embedding resins that enable histological, immunohistochemical, and morphological examination. However, the suitability of PMMA-embedded hard tissues for large-scale proteomic analysis remained unexplored. This study is the first to report on the feasibility of PMMA-embedded bone samples for LC-MS/MS analysis. Conventional workflows yielded merely limited coverage of the bone proteome. Using advanced strategies of prefractionation by high-pH reversed-phase liquid chromatography in combination with isobaric tandem mass tag labeling resulted in proteome coverage exceeding 1000 protein identifications. The quantitative comparison with cryopreserved samples revealed that each sample preparation workflow had a distinct impact on the proteomic profile. However, workflow replicates exhibited a high reproducibility for PMMA-embedded samples. Our findings further demonstrate that decalcification prior to protein extraction, along with the analysis of solubilization fractions, is not preferred for PMMA-embedded bone. The biological applicability of the proposed workflow was demonstrated using samples of human PMMA-embedded alveolar bone and the iliac crest, which revealed anatomical site-specific proteomic profiles. Overall, these results establish a crucial foundation for large-scale proteomics studies contributing to our knowledge of bone biology.

In vitro assessment of internal implant-abutment connections with different cone angles under static loading using synchrotron-based radiation.

BACKGROUND: The stability of implant-abutment connection is crucial to minimize mechanical and biological complications. Therefore, an assessment of the microgap behavior and abutment displacement in different implant-abutment designs was performed. METHODS: Four implant systems were tested, three with a conical implant-abutment connection based on friction fit and a cone angle < 12 ° (Medentika, Medentis, NobelActive) and a system with an angulated connection (< 40°) (Semados). In different static loading conditions (30 N - 90º, 100 N - 90º, 200 N - 30º) the microgap and abutment displacement was evaluated using synchrotron-based microtomography and phase-contrast radioscopy with numerical forward simulation of the optical Fresnel propagation yielding an accuracy down to 0.1 µm. RESULTS: Microgaps were present in all implant systems prior to loading (0.15-9 µm). Values increased with mounting force and angle up to 40.5 µm at an off axis loading of 100 N in a 90° angle. CONCLUSIONS: In contrast to the implant-abutment connection with a large cone angle (45°), the conical connections based on a friction fit (small cone angles with < 12°) demonstrated an abutment displacement which resulted in a deformation of the outer implant wall. The design of the implant-abutment connection seems to be crucial for the force distribution on the implant wall which might influence peri-implant bone stability.

Mass spectrometry-based proteomic applications in dental implants research.

Dental implants have been established as successful treatment options for missing teeth with steadily increasing demands. Today, the primary areas of research in dental implantology revolve around osseointegration, soft and hard tissue grafting as well as peri-implantitis diagnostics, prevention, and treatment. This review provides a comprehensive overview of the current literature on the application of MS-based proteomics in dental implant research, highlights how explorative proteomics provided insights into the biology of peri-implant soft and hard tissues and how proteomics facilitated the stratification between healthy and diseased implants, enabling the identification of potential new diagnostic markers. Additionally, this review illuminates technical aspects, and provides recommendations for future study designs based on the current evidence.

Comparison of the 3D-microstructure of human alveolar and fibula bone in microvascular autologous bone transplantation: a synchrotron radiation µ-CT study.

Introduction: Autologous bone transplantation is successfully used in reconstructive surgery of large/critical-sized bone defects, whereby the microvascular free fibula flap is still regarded as the gold standard for the reconstruction of such defects in the head and neck region. Here, we report the morphological and lacunar properties of patient-paired bone samples from eight patients from the jaw (AB; recipient site) and the fibula (FB; donor site) on the micron length-scale using Synchrotron µ-CT. Insights into differences and similarities between these bone structures could offer a better understanding of the underlying mechanism for successful surgical outcomes and might clear the path for optimized, nature-inspired bone scaffold designs. Methods: Spatial vessel-pore arrangements, bone morphology, fluid-simulation derived permeability tensor, osteocyte lacunar density, and lacunar morphology are compared. Results: The orientation of the vessel system indicates a homogenous vessel orientation for AB and FB. The average mineral distance (50%) to the closest vessel boundary is higher in AB than in FB (the mean is 96 µm for AB vs. 76 µm for FB; p = 0.021). Average osteocyte lacunar density is found to be higher in AB than in FB (mean 22,874 mm3 vs. 19,376 mm3 for FB; p = 0.038), which might compensate for the high distance from the mineral to the nearest vessel. No significant differences in lacunar volume are found between paired AB and FB. Discussion: A comparable vessel network and similar distribution of vessel porosity between AB and FB may allow the FB graft to exhibit a high regeneration potential when connected to AB, and this might correlate with a high osteoinductive and osteoconductive potential of FB when connected to AB. Since widely used and potent synthetic bone grafts exist, new insight into the bone structure of well-established autologous bone grafts, such as the free fibula flap, could help to improve the performance of such materials and therefore the design of 3D scaffolds.

Root Coverage Using a Novel Porcine Acellular Dermal Matrix: Case Reports of Different Minimally Invasive Techniques with a 3-Year Follow-up.

A recently released commercially available novel porcine acellular dermal matrix (PADM) appears to possess acceptable biologic and clinical properties to be considered as an acceptable soft tissue replacement material. The aim of these three case reports is to present the treatment of multiple gingival recession by means of different variations of the tunnel and PADM as well as the clinical outcomes obtained at 3 years postoperatively. The healing outcomes demonstrated only minor surgical complications, with minimal patient-reported discomfort. At 3 years postoperative, ideal functional and esthetic outcomes were observed. PADM seems to be a promising xenogeneic soft tissue substitute. Further studies with a higher number of patients and defects are necessary to confirm the present findings.

Impact of peri-implantitis on the proteome biology of crevicular fluid: A pilot study.

BACKGROUND: The proteome of the peri-implant crevicular fluid (PICF) has not been systematically investigated. The aim of the present study was to reveal the proteome biology of dental implants affected with peri-implantitis. METHODS: Patients with at least one diseased implant were included (probing depth ≥6 mm, ≥3 mm peri-implant radiological bone loss). Using sterile paper strips, samples were collected from healthy implants (I), healthy teeth (T) and peri-implantitis affected implants (P). Proteome analysis was performed using liquid chromatography - tandem mass spectrometry (LC-MS/MS) and data independent acquisition, allowing the identification and quantification of human and bacterial proteins as well as semi-specific peptides. RESULTS: A total of 38 samples from 14 patients were included in the study; 2332 different human proteins were identified across all samples. No differentially expressed proteins between T and I were found. Comparing P to I, 59 proteins were found upregulated and 31 downregulated in P with significance. Upregulated proteins included proinflammatory proteins such as immunoglobulins, dysferlin, and S100P, as well as antimicrobial proteins, for example, myeloperoxidase or azurocidin. Gene ontology analysis further revealed higher activity of immunological pathways. Proteolytic patterns indicated the activity of inflammatory proteins such as cathepsin G. A total of 334 bacterial proteins were identified and quantified. Peri-implantitis showed elevated proteolytic activity. CONCLUSION: I and T share similarities in their proteome, while diseased implants deviate strongly from healthy conditions. The PICF proteome of peri-implantitis affected sites exhibits an inflammatory fingerprint, dominated by neutrophil activity when compared with healthy implants.

Association of sex steroid hormones and new bone formation rate after iliac onlay grafting: a prospective clinical pilot study.

PURPOSE: The present prospective study evaluates the association between new bone formation rate in the iliac onlay graft and sex steroid hormone serum levels. METHODS: A total of 15 partially or completely edentulous postmenopausal females and 9 males with less than 5 mm height of the remaining alveolar bone underwent iliac onlay grafting followed by dental implant placement using a two-stage approach. Sex hormone binding globulin and 17ß-estradiol serum levels were investigated by electrochemiluminescence immunoassay, while total testosterone level was analyzed using radioimmunoassay. At the time of implant placement, 12 weeks after grafting, bone biopsies were obtained and analyzed histomorphometrically. Statistical analysis was performed using linear mixed models. RESULTS: Grafting procedure was successfully performed in all patients. The mean new bone formation rate was 32.5% (116 samples). In men the mean new bone formation rate (38.1%) was significantly higher (p < 0.01) than in women (27.6%). Independent of gender 17ß-estradiol and testosterone were positively associated to overall new bone formation rate, albeit a significant influence was only seen for 17ß-estradiol in men (p = 0.020). Sex hormone binding globulin had no influence on new bone formation rate (p = 0.897). There was no significant association between new bone formation rate and age (p = 0.353) or new bone formation rate and body mass index (p = 0.248). CONCLUSION: Positive association of 17ß-estradiol as well as testosterone with new bone formation rate after iliac onlay grafting indicates a role of sex steroid hormones in alveolar bone regeneration, although the observed influence was only significant for 17ß-estradiol in men.

Impurities in commercial titanium dental implants - A mass and optical emission spectrometry elemental analysis.

OBJECTIVE: Titanium (Ti) is considered bioinert and is still regarded as the "gold standard" material for dental implants. However, even 'commercial pure' Ti will contain minor fractions of elemental impurities. Evidence demonstrating the release of Ti ions and particles from 'passive' implant surfaces is increasing and has been attributed to biocorrosion processes which may provoke immunological reactions. However, Ti observed in peri-implant tissues has been shown to be co-located with elements considered impurities in biomedical alloys. Accordingly, this study aimed to quantify the composition of impurities in commercial Ti dental implants. METHODS: Fifteen commercial titanium dental implant systems were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS) and optical emission spectrometry (ICP-OES). RESULTS: The elemental composition of implants manufactured from commercially pure grades of Ti, Ti-6Al-4V, and the TiZr alloy (Roxolid) conformed to the respective ISO/ASTM standards or manufacturers´ data (TiZr/Roxolid). However, all implants investigated included exogenous metal contaminants including Ni, Cr, Sb, and Nb to a variable extent. Other contaminants detected in a fraction of implants included As and the radionuclides U-238 and Th-232. SIGNIFICANCE: Although all Ti implant studies conformed with their standard compositions, potentially allergenic, noxious metals and even radionuclides were detected. Since there are differences in the degree of contamination between the implant systems, a certain impurity fraction seems technically avoidable. The clinical relevance of these findings must be further investigated, and an adaptation of industry standards should be discussed.

Comparison of the 3D-Microstructure Between Alveolar and Iliac Bone for Enhanced Bioinspired Bone Graft Substitutes.

In oral- and maxillofacial bone augmentation surgery, non-vascularized grafts from the iliac crest demonstrate better clinical performance than alveolar bone grafts. The underlying mechanisms are not fully understood but are essential for the enhancement of bone regeneration scaffolds. Synchrotron Radiation µ-CT at a pixel size of 2.3 µm was used to characterize the gross morphology and the vascular and osteocyte lacuna porosity of patient-matched iliac crest/alveolar bone samples. The results suggest a difference in the spatial distribution of the vascular pore system. Fluid simulations reveal the permeability tensor to be more homogeneous in the iliac crest, indicating a more unidirectional fluid flow in alveolar bone. The average distance between bone mineral and the closest vessel pore boundary was found to be higher in alveolar bone. At the same time, osteocyte lacunae density is higher in alveolar bone, potentially compensating for the longer average distance between the bone mineral and vessel pores. The present study comprehensively quantified and compared the 3D microarchitecture of intraindividual human alveolar and iliac bone. The identified difference in pore network architecture may allow a bone graft from the iliac crest to exhibit higher regeneration potential due to an increased capacity to connect with the surrounding pore network of the residual bone. The results may contribute to understanding the difference in clinical performance when used as bone grafts and are essential for optimization of future scaffold materials.

Proteomic profiling of human bone from different anatomical sites - A pilot study.

PURPOSE: The study aim is a comparative proteome-based analysis of different autologous bone entities (alveolar bone [AB], iliac cortical [IC] bone, and iliac spongiosa [IS]) used for alveolar onlay grafting. EXPERIMENTAL DESIGN: Site-matched bone samples of AB, IC, and IS were harvested during alveolar onlay grafting. Proteins were extracted using a detergent-based (sodium dodecyl sulfate) strategy and trypsinized. Proteome analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). MaxQuant was used for peptide-to-spectrum matching, peak detection, and quantitation. Linear models for microarray analysis (LIMMA) were used to detect differentially abundant peptides and proteins. RESULTS: A total of 1730 different proteins were identified across the 15 samples at a false discovery rate of 1%. Partial least-squares discriminant analysis approved segregation of AB, IC, and IS protein profiles. LIMMA statistics highlighted 66 proteins that were more abundant in AB then in IC (vs. 92 proteins were enriched in IC over AB). Gene Ontology enrichment analysis revealed a matrisomal versus an immune-related proteome fingerprint in AB versus IC. CONCLUSION AND CLINICAL RELEVANCE: This pilot study demonstrates an ECM protein-related proteome fingerprint in AB and an immune-related proteome fingerprint in IS and IC.

Efficacy of alternative or adjunctive measures to conventional non-surgical and surgical treatment of peri-implant mucositis and peri-implantitis: a systematic review and meta-analysis.

PURPOSE: To evaluate the efficacy of alternative or adjunctive measures to conventional non-surgical or surgical treatment of peri-implant mucositis and peri-implantitis. MATERIAL AND METHODS: Prospective randomized and nonrandomized controlled studies comparing alternative or adjunctive measures, and reporting on changes in bleeding scores (i.e., bleed0ing index (BI) or bleeding on probing (BOP)), probing depth (PD) values or suppuration (SUPP) were searched. RESULTS: Peri-implant mucositis: adjunctive use of local antiseptics lead to greater PD reduction (weighted mean difference (WMD) = - 0.23 mm; p = 0.03, respectively), whereas changes in BOP were comparable (WMD = - 5.30%; p = 0.29). Non-surgical treatment of peri-implantitis: alternative measures for biofilm removal and systemic antibiotics yielded higher BOP reduction (WMD = - 28.09%; p = 0.01 and WMD = - 17.35%; p = 0.01, respectively). Surgical non-reconstructive peri-implantitis treatment: WMD in PD amounted to - 1.11 mm favoring adjunctive implantoplasty (p = 0.02). Adjunctive reconstructive measures lead to significantly higher radiographic bone defect fill/reduction (WMD = 56.46%; p = 0.01 and WMD = - 1.47 mm; p = 0.01), PD (- 0.51 mm; p = 0.01) and lower soft-tissue recession (WMD = - 0.63 mm; p = 0.01), while changes in BOP were not significant (WMD = - 11.11%; p = 0.11). CONCLUSIONS: Alternative and adjunctive measures provided no beneficial effect in resolving peri-implant mucositis, while alternative measures were superior in reducing BOP values following non-surgical treatment of peri-implantitis. Adjunctive reconstructive measures were beneficial regarding radiographic bone-defect fill/reduction, PD reduction and lower soft-tissue recession, although they did not improve the resolution of mucosal inflammation.

Cross-sectional evaluation of clinical and immunological parameters at partially microgrooved vs machined abutments in humans.

OBJECTIVE: The objective of the present study was to examine the clinical and immunological parameters in samples collected from the peri-implant crevicular fluid (PICF) of machined titanium (M) abutments compared to titanium abutments with a laser-microtextured surface (LMS) on dental implants. MATERIAL AND METHODS: A total of 40 patients with one titanium implant, half of them (n=20) provided with a M abutment (control group) and the other half (n=20) with LMS abutments (test group), were included in the study. Clinical parameters pocket probing depth (PD), full-mouth plaque score (FMPS), radiographic bone loss (RBL), clinical attachment level (CAL), mucosal recession (MR), bleeding on probing (BOP), and width of keratinized mucosa (KM) were evaluated. The peri-implant sulcus fluid was analyzed for cytokines IL-1α, IL-1ß, IL-6, IL-8, and IL-10 via flow cytometry. RESULTS: Clinical evaluation demonstrated no significant difference of PD (mean LMS = 3.50 mm/SD 0.95 mm vs mean M = 3.45 mm/SD 0.76 mm (p=0.855)), MR (mean LMS = 0.30 mm/SD 0.57 mm vs mean M = 0.35 mm/SD 0.67 mm (p=0.801)), CAL (mean LMS = 3.60 mm/SD 1.14 mm vs mean M = 3.55 mm/SD 0.89 mm (p=0.878)), and KM (mean LMS = 2.03 mm/SD 1.08 mm vs mean M = 2.13 mm/SD 0.92 mm (p=0.754)) between LMS and M abutments. LMS abutments showed less BOP than M abutments (26.7% vs 30.8%), but statistically not significant (p = 0.2235). Radiographic bone loss (mean LMS = 0.22 mm/SD 0.44 mm vs mean M = 0.59 mm/SD 0.49 mm) was reduced in the test group in comparison with the control group (p=0.016). In the collected PICF, the levels of pro-inflammatory cytokines IL-1α (median LMS = 180.8 pg/ml vs M = 200.9 pg/ml (p=0.968)) and IL-1ß (median LMS = 60.43 pg/ml vs M = 83.11 pg/ml (p=0.4777)) were lower, and the levels of IL-6 (median LMS = 180.8 pg/ml vs M = 200.9 pg/ml (p<0.0001)) were significantly lower in the test group. In contrast, the levels of IL-8 (median LMS = 255.7 pg/ml vs M = 178.7 pg/ml (p=0.3306)) were higher in the test group, though not significantly. The levels of anti-inflammatory IL-10 were significantly increased in the test group (LMS median = 0.555 pg/ml vs M median = 0.465 pg/ml (p=0.0365)). IL-1ß showed a significant correlation to radiologic bone loss (p=0.0024). The other variables IL-1α, IL-6, IL-8, and IL-10 had no significant correlation to radiological bone loss. CONCLUSION: Within the limitations of this study, titanium implants provided with laser-microtextured surface abutments seem to demonstrate less pro-inflammatory and more anti-inflammatory activity and to show reduced radiographic bone loss compared to machined titanium abutments. CLINICAL RELEVANCE: The use of laser-microtextured surface abutments might have the potential to support peri-implant tissue health.

Recommendations for Implant-Supported Full-Arch Rehabilitations in Edentulous Patients: The Oral Reconstruction Foundation Consensus Report.

The tasks of Working Groups 1 to 6 at the 4th Consensus Meeting of the Oral Reconstruction Foundation were to elucidate clinical recommendations for implant-supported full-arch rehabilitations in edentulous patients. Six systematic/narrative reviews were prepared to address the following subtopics: (1) the influence of medical and geriatric factors on implant survival; (2) the prevalence of peri-implant diseases; (3) the influence of material selection, attachment type, interarch space, and opposing dentition; (4) different interventions for rehabilitation of the edentulous maxilla; (5) different interventions for rehabilitation of the edentulous mandible; and (6) treatment choice and decision-making in elderly patients. Consensus statements, clinical recommendations, and implications for future research were determined based on structured group discussions and plenary session approval.

Influence of Medical and Geriatric Factors on Implant Success: An Overview of Systematic Reviews.

PURPOSE: To provide an overview of the influence of medical and geriatric factors on implant survival in order to form clinical recommendations for the practitioner. MATERIALS AND METHODS: This narrative literature review was performed to address the following questions: (1) Is age (> 75 years) a risk factor for implant survival?; (2) Is diabetes mellitus a risk factor for implant survival?; and (3) Is antiresorptive therapy a risk factor for implant survival? The PubMed, Web of Knowledge (Thomson Reuters), and Google Scholar databases were searched for systematic reviews and research papers of evidence level II and above that were published up to February 2019 for each topic. RESULTS: (1) Age > 75 years does not affect implant survival according to short-term follow up (1 to 5 years). However, polypharmacy should be considered in this patient group. (2) Diabetes mellitus is not a risk factor for implant survival in the short term, but there is no information on appropriate perioperative treatment and wound closure. There is little evidence in the literature on the success of bone grafting and progressive loading protocols in diabetic patients. (3) Implant therapy cannot be recommended in patients under high-dose bisphosphonate and antibody therapy. Bone grafting should be avoided under antiresorptive therapy. There are no treatment regimens available for patients with peri-implantitis receiving antiresorptive medication. CONCLUSION: This review suggests that the risk assessment for an implant patient should not be based on age, but rather on the patient's specific risk factors, such as former and current diseases and medication.

Immunohistological composition of peri-implantitis affected tissue around ceramic implants-A pilot study.

BACKGROUND: Aim of the pilot study was the histologic classification of the inflamed peri-implant soft tissue around ceramic implants (CI) in comparison with titanium implants (TI). METHODS: Peri-implant tissue were retrieved from 15 patients (aged 34 to 88 years, seven males/eight females) with severe peri-implantitis (eight CI, seven TI). The peri-implant soft tissue samples were retrieved from the sites during scheduled removal of the implant and prepared for immunohistochemical analysis. Monoclonal antibodies (targeting CD3, CD20, CD138, and CD68) were used to identify T- and B-cells, plasma cells and macrophages. Quantitative assessment was performed by one histologically trained investigator. Linear mixed regression models were used. RESULTS: A similar numerical distribution of the cell population was found in peri-implantitis around CI compared with TI. CD3 (TI, 17% to 85% versus CI, 20% to 70% of total cell number) and CD138 (TI, 1% to 73% versus CI, 12% to 69% of total cell number) were predominantly expressed. Notably, patient-individual differences of numerical cell distribution were detected. Co-localization of B- and T-lymphocytes was observed. CONCLUSIONS: Peri-implantitis around CI in comparison with TI seems to have a similar histological appearance. Differences in cellular composition of peri-implantitis lesions might also depend on the patient's specific immune status and not only on the material used.

Distribution and Chemical Speciation of Exogenous Micro- and Nanoparticles in Inflamed Soft Tissue Adjacent to Titanium and Ceramic Dental Implants.

Degradation of the implant surface and particle release/formation as an inflammation catalyst mechanism is an emerging concept in dental medicine that may help explain the pathogenesis of peri-implantitis. The aim of the present study was a synchrotron-based characterization of micro- and nanosized implant-related particles in inflamed human tissues around titanium and ceramic dental implants that exhibited signs of peri-implantitis. Size, distribution, and chemical speciation of the exogenous micro- and nanosized particle content were evaluated using synchrotron µ-X-ray fluorescence spectroscopy (XRF), nano-XRF, and µ-X-ray absorption near-edge structure (XANES). Titanium particles, with variable speciation, were detected in all tissue sections associated with titanium implants. Ceramic particles were found in five out of eight tissue samples associated with ceramic implants. Particles ranged in size from micro- to nanoscale. The local density of both titanium and ceramic particles was calculated to be as high as ∼40 million particles/mm3. µ-XANES identified titanium in predominantly two different chemistries, including metallic and titanium dioxide (TiO2). The findings highlight the propensity for particle accumulation in the inflamed tissues around dental implants and will help in guiding toxicological studies to determine the biological significance of such exposures.

Horizontal bone grafting using equine-derived cancellous bone blocks is associated with severe complications: A prospective clinical and histological pilot study.

AIMS: The aim of this prospective, clinical study was to evaluate the clinical performance and histological outcome of a new equine hydroxyapatite collagenated bone block (eHAC) for horizontal bone grafting prior to implant placement. MATERIALS AND METHODS: Five patients (two male/three female) with a mean age of 51.6 years (range 22-66 years) and a reduced horizontal bone width of the alveolar ridge (mean 3.5 mm) underwent horizontal bone grafting using eHAC at 10 grafting sites. Reentry was performed 6.9 months after the horizontal grafting procedure. Clinical follow-up (mean 28.9 month) considered width gain of the alveolar ridge, soft tissue healing, and complications. To evaluate graft incorporation, four additional patients underwent histological assessment of equine blocks adjacent to autologous blocks 3 and 6 months after grafting. RESULTS: The study was terminated after graft failure was observed in four of five patients. Mean horizontal bone width had increased by 3.6 ± 1.22 mm. Three out of nine implants placed had to be removed due to graft failure. Histological evaluation revealed large amounts of soft connective tissue within the grafts (mean 67.3 ± 9.5%). The proportion of new bone formation 3 months after the lateral grafting procedure revealed an average of 8.6%, compared to 11.4% after 6 to 7 months. CONCLUSION: Lateral ridge grafting using eHAC achieved measurable horizontal width gain but revealed high rates of severe complications. CLINICAL IMPLICATIONS: Within the limitations of this study, eHAC bone blocks cannot be recommended for horizontal bone grafting.

High Potential of Bacterial Adhesion on Block Bone Graft Materials.

Bone graft infections represent a challenge in daily clinics, resulting in increased patient discomfort and graft removal. The aim of this study was to investigate the initial adhesion of five representative pathogens on three different block bone graft materials (xenogeneic, alloplastic and allogeneic) and to assess if chlorhexidine (CHX) can effectively control the initial bacterial adhesion. Three different block bone grafting materials (Tutobone®, Endobon® and human spongiosa) were incubated with Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis and Pseudomonas aeruginosa in the presence or absence of 0.2% CHX solution. Bacterial adhesion was assessed by the direct counting of the colony-forming units (CFUs) and visualized by scanning electron microscopy (SEM). Overall, the selected bacterial species adhered successfully to all tested bone replacement scaffolds, which showed similar bacterial counts. The lg CFU values ranged from 5.29 ± 0.14 to 5.48 ± 0.72 for E. coli, from 4.37 ± 0.62 to 5.02 ± 0.48 for S. aureus, from 4.92 ± 0.34 to 4.95 ± 0.21 for S. mutans, from 4.97 ± 0.40 to 5.22 ± 0.13 for E. faecalis and from 4.23 ± 0.54 to 4.58 ± 0.26 for P. aeruginosa. CHX did not interfere with initial microbial adhesion, and yet it killed all adhered bacterial cells. Thus, CHX can be used to prevent subsequent biofilm infections.

Elemental analysis of commercial zirconia dental implants - Is "metal-free" devoid of metals?

OBJECTIVES: The interest in ceramic dental implants made of yttria-stabilized tetragonal zirconia polycrystals (Y-TZP) or alumina toughened zirconia (ATZ) has increased in recent years. However, in the light of aging, corrosion, and potential impurities of zirconia ceramics, the material composition of these implants and the associated term "metal-free" is persistently questioned. Thus, the present study aimed to conduct an elemental analysis of commercial zirconia dental implants to specify their elemental composition and to identify contaminants. METHODS: Nine commercial zirconia dental implant systems and corresponding material samples were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS) and optical emission spectrometry (ICP-OES). RESULTS: While the elemental composition was dominated by the main components Zr, Y and Al (in ATZ samples), all investigated samples contained impurities with Hf and contamination with alkali and alkali earth elements (Na, K, Mg, Ca), essential trace elements (e.g. Fe, Cu, Zn) but also potentially noxious metal elements (e.g. Ni, Cr). Furthermore, ultra-trace level contamination with the radionuclides U-238 and Th-232 was found in the majority of samples. SIGNIFICANCE: The results indicate that, although all the investigated Y-TZP and ATZ dental implants meet the currently relevant ISO standards and manufacturer's specifications, from an elemental point of view, they are not devoid of metals. Due to the lack of a universal definition and thresholds for the term "metal-free", the question of whether the examined zirconia dental implants can be holistically classified as "metal-free" or not remains a controversial, philosophical one.

3-D Printed Protective Equipment during COVID-19 Pandemic.

While the number of coronavirus cases from 2019 continues to grow, hospitals are reporting shortages of personal protective equipment (PPE) for frontline healthcare workers. Furthermore, PPE for the eyes and mouth, such as face shields, allow for additional protection when working with aerosols. 3-D printing enables the easy and rapid production of lightweight plastic frameworks based on open-source data. The practicality and clinical suitability of four face shields printed using a fused deposition modeling printer were examined. The weight, printing time, and required tools for assembly were evaluated. To assess the clinical suitability, each face shield was worn for one hour by 10 clinicians and rated using a visual analogue scale. The filament weight (21-42 g) and printing time (1:40-3:17 h) differed significantly between the four frames. Likewise, the fit, wearing comfort, space for additional PPE, and protection varied between the designs. For clinical suitability, a chosen design should allow sufficient space for goggles and N95 respirators as well as maximum coverage of the facial area. Consequently, two datasets are recommended. For the final selection of the ideal dataset to be used for printing, scalability and economic efficiency need to be carefully balanced with an acceptable degree of protection.