Autoclaving-induced dimensional changes of three-dimensional printed surgical guides: An in vitro study.

OBJECTIVES: Surgical guides are frequently used for dental implant placement. The aim of this study was to evaluate the impact of the 3D printing process itself and subsequent steam autoclaving on the dimensional stability of five different resin/printer combinations (RPCs). MATERIALS AND METHODS: Fifty identical surgical guides (10 per group) were produced consisting of five RPCs. Half of the guides (5 per group) were steam autoclaved with cycle 1 (121°C, 1 bar, 20.5 min) and the other half with cycle 2 (134°C, 2 bar, 5.5 min). All guides were scanned with a structured-light (SL) 3D scanner before (T0) and after (T1) autoclaving. Linear measurements along the x-, y-, and z-axes were performed at landmarks on the original STL file and on SL scans at T0 and T1, respectively. Wilcoxon signed-rank test, Kruskal-Wallis test, and linear mixed-effects models were performed, depending on the analysis. RESULTS: Three-dimensional printing was associated with significant dimensional alterations for all RPCs. Steam autoclaving using cycle 1 was associated with significant shrinkage in x- (1 RPC), y- (2 RPCs), and z-direction (2 RPCs), while cycle 2 was also associated with shrinkage in x- (2 RPCs), y- (1 RPC), and z-direction (1 RPC). One resin did not present any dimensional changes independently of the cycle. CONCLUSIONS: The majority of the guides presented minor but significant shrinkage due to 3D printing itself and both steam autoclaving cycles, the extent varied between different RPCs. Whether these changes compromise implant placement accuracy remains to be investigated.

Influence of antiresorptive/antiangiogenic therapy on the extension of experimentally induced peri-implantitis lesions.

OBJECTIVES: To investigate the extension of experimentally induced peri-implantitis lesions under various antiresorptive and antiangiogenic medications. MATERIAL AND METHODS: Fourty-eight albino rats had randomly received the following medications (dual application, n = 8 each): (1) amino-bisphosphonate (zoledronate) (Zo), (2) RANKL inhibitor (denosumab) (De), (3) antiangiogenic (bevacizumab) (Be), (4) Zo+Be, (5) De+Be, or (6) no medication (Co). Ligature- and lipopolysaccharide-induced peri-implantitis lesions were established at 2 maxillary implants over a period of 16 weeks. Histological (e.g., apical extension and surface area of the inflammatory cell infiltrate-aICT, ICT; defect length; defect width; CD68 positive cells) and bone micromorphometric (µCT) outcomes were assessed. The animal was defined as a statistical unit. RESULTS: A total of n = 38 animals (Zo = 6, De = 6, Be = 8, Zo + Be = 6, De + Be = 5, Co = 7) were analyzed. ICT's were commonly marked by a positive CD68 antigen reactivity. Comparable median aICT (lowest-Zo: 0.53 mm; highest-Be: 1.22 mm), ICT (lowest-De + Be: 0.00 mm2; highest-Co: 0.49 mm2), defect length (lowest-Zo: 0.90 mm; highest-Co: 1.93 mm) and defect width (lowest-De+Be: 1.27 mm; highest-Be: 1.80 mm) values were noted in all test and control groups. Within an inner (diameter: 0.8 mm) cylindric volume of interest, the bone microstructure did not significantly differ between groups. CONCLUSIONS: The present analysis did not reveal any marked effects of various antiresorptive/ antiangiogenic medications on the extension of experimentally induced peri-implantitis lesions. CLINICAL RELEVANCE: The extension of peri-implantitis lesions may not be facilitated by the antiresorptive and antiangiogenic medications investigated.

Micro-angiogenic patterns around orthodontic implants migrating in bone: A micro-CT study in the rat tail model.

AIM: Recent studies revealed that implants can migrate in bone when subjected to continuous loading. Since this process is suspected to be accompanied by bone remodelling, which requires blood vessel formation, the present work aimed at assessing the micro-angiogenic patterns around migrating implants. MATERIALS AND METHODS: In 16 rats, two customized implants were placed in a single tail vertebra and connected with contraction springs (forces: 0 N, 0.5 N, 1.0 N, 1.5 N). After 2 or 8 weeks of loading, the animals were scanned by micro-CT before and after vasculature perfusion with a silicone rubber. Vessels were segmented by subtraction of the two micro-CT scans. Vessel thickness (V.Th), vessel volume per total volume (VV/TV), and vascular spacing (V.Sp) were assessed in a peri-implant volume of interest (VOI) around each implant. RESULTS: At 2 weeks of loading, force magnitude was significantly associated with VV/TV and V.Th values (χ2  = 10.942, p < .001 and χ2  = 6.028, p = .010, respectively). No significant differences were observed after 8 weeks of loading. CONCLUSIONS: Within the limitations of an animal study, peri-implant vessel thickness and density were associated with force magnitude in the early loading phase, whereas effects diminished after 8 weeks of loading.

Two-piece zirconia implants in the posterior mandible and maxilla: A cohort study with a follow-up period of 9 years.

OBJECTIVES: Long-term follow-up observations of zirconia implants are rare. This study aimed at evaluating the clinical performance of two-piece zirconia implants in the posterior jaws over 9 years. MATERIALS AND METHODS: Sixty partially edentulous patients were treated with two-piece zirconia implants. In eight no primary stability could be achieved. Fifty-two patients received the final restoration (i.e., cemented fibreglass abutments and all-ceramic crowns). After 2 years, 2 implants failed and 4 dropouts were recorded. The remaining 46 patients with one target implant each were recalled at 9 years. Besides implant survival, clinical parameters at the implant level (plaque index-PI, bleeding on probing-BOP, probing depth-PD, mucosal recession-MR) were recorded and compared with previously collected data. Mechanical and technical complications were assessed. RESULTS: Thirty patients responded. The mean observation period was of 111.1 ± 2.2 months. One implant was lost. Data recorded from the remaining 29 implants were analysed. PI values increased overtime. Mean BOP and PD remained unchanged during follow-up. No additional cases of peri-implantitis were recorded over the 10 diagnosed during the first 2 years of follow-up. No significant changes in mean MR values were detected over time, with 65% of the all included implants exhibiting no recession at 9 years and all the others, but one, a maximum MR of 1 mm. Three technical and 6 mechanical complications occurred in 7 patients between 2- and 9-years (6.9% and 20.7%, respectively, at patient level). CONCLUSION: Within the limitations of the present study, a high survival rate was registered. Albeit frequent mechanical and technical complications, two-piece zirconia implants could represent a valid solution for the replacement of single teeth in the posterior jaws.

Volumetric analysis of the periodontal microstructure under antiresorptive therapy: an experimental study in rabbits.

OBJECTIVES: The objective of this study was to volumetrically assess changes in the periodontal microstructure under antiresorptive therapy. MATERIALS AND METHODS: Microtomographic scans from a total of 9 Dutch Belted rabbits having been randomly allocated to either the intravenous administration of amino-bisphosphonate (zoledronic acid) (Za) (n = 5) or a negative control group (nZa) (n = 4) were obtained at 10 months following a repeated drug administration. A quantification of the periodontal space thickness (P.Th) of both maxillary and mandibular most posterior premolars, as well as of the 2nd molars was performed. Bone micromorphometry was assessed by means of bone volume per total volume (BV/TV), the bone mineral density (BMD), trabecular thickness (Tb.Th), trabecular number (Tb.N), bone surface (BS), and the specific bone surface (BS/BV). RESULTS: Za was associated with significantly higher P.Th (P = 0.010), which was most pronounced in the upper jaw. Bone micromorphometry revealed no significant differences among the two groups, i.e., Za and nZa, for all the investigated parameters. CONCLUSIONS: Volumetric analysis revealed that antiresorptive therapy was associated with periodontal space widening, whereas major effects on the bone micro-morphology could not be observed. CLINICAL RELEVANCE: A deep understanding of specific periodontal and alveolar bone alterations in patients under antiresorptive therapy might help to prevent the onset of medication-related osteonecrosis of the jaw.

Bone remodelling patterns around orthodontic mini-implants migrating in bone: an experimental study in rat vertebrae.

BACKGROUND: Orthodontic implant migration has been clinically observed in presence of continuous loading forces. Recent studies indicate that osteocytes play a crucial role in this phenomenon. OBJECTIVES: Aim of this study was to investigate local osteocytic gene expression, protein expression, and bone micro-structure in peri-implant regions of pressure and tension. MATERIAL AND METHODS: The present work reports a complementary analysis to a previous micro-computed tomography study. Two customized mini-implants were placed in one caudal rat vertebra and connected by a nickel-titanium contraction spring generating different forces (i.e. 0, 0.5, 1.0, and 1.5 N). Either at 2 or 8 weeks, the vertebrae were harvested and utilized for 1. osteocytic gene expression using laser capture micro-dissection on frozen sections coupled with qPCR, 2. haematoxylin-eosin staining for qualitative and quantitative analyses, 3. immunofluorescence staining and analysis, and 4. bone-to-implant contact on undecalcified samples. RESULTS: At the two time points for all the performed analyses no significant differences were observed with respect to the applied force magnitudes and cell harvesting localization. However, descriptive histological analysis revealed remarkable bone remodelling at 2 weeks of loading. At 8 weeks the implants were osseointegrated and, especially in 1.0 and 1.5 N groups, newly formed bone presented a characteristic load bearing architecture with trabecula oriented in the direction of the loading. CONCLUSIONS: The present study confirmed that stress-induced bone remodelling is the biological mechanism of orthodontic implant migration. Bone apposition was found at 'tension' and 'pressure' sites thus limiting implant migration over time.

Can implants move in bone? A longitudinal in vivo micro-CT analysis of implants under constant forces in rat vertebrae.

OBJECTIVES: Whereas stationary stability of implants has been postulated for decades, recent studies suggested a phenomenon termed implant migration. This describes a change in position of implants as a reaction to applied forces. The present study aims at employing image registration of in vivo micro-CT scans from different time points and to assess (a) if migration of continuously loaded implants is possible and (b) migration correlates with the force magnitude. MATERIAL AND METHODS: Two customized machined implants were placed in the dorsal portion of caudal vertebrae in n = 61 rats and exposed to standardized forces (0.5 N, 1.0 N, and 1.5 N) applied through a flat nickel-titanium contraction spring, or no forces (control). Micro-CT scans were performed at 0, 1, 2, 4, 6, and 8 weeks after surgery. The baseline image was registered with the forthcoming scans. Implant migration was measured as the Euclidean distance between implant tips. Bone remodeling was assessed between the baseline and the forthcoming scans. RESULTS: The findings confirmed a positional change of the implants at 2 and 8 weeks of healing, and a linear association between applied force and velocity of movement (anterior implant: χ2  = 12.12, df = 3, and p = .007 and posterior implant: χ2  = 20.35, df = 3, and p < .001). Bone apposition was observed around the implants and accompanied by formation of load-bearing trabeculae and a general cortical thickening close and also distant to the implants. CONCLUSION: The present analysis confirmed that implants can migrate in bone. The applied forces seemed to stimulate bone thickening, which could explain why implants migrate without affecting stability.

Application of Piezo-Based Measuring System for Evaluation of Nucleic Acid-Based Drugs Influencing the Coagulation.

During open-heart surgery, the status of hemostasis has to be constantly monitored to quickly and reliably detect bleeding or coagulation disorders. In this study, a novel optimized piezo-based measuring system (PIEZ) for rheological monitoring of hemostasis was established. The applicability of the PIEZ for the evaluation of nucleic acid-based drugs influencing coagulation was analyzed. Thrombin aptamers such as NU172 might be used during extracorporeal circulation (ECC) in combination with a reduced heparin concentration or for patients with heparin-induced thrombocytopenia (HIT). Therefore, the effect of the coagulation inhibiting thrombin aptamer NU172 and the abrogation by its complementary antidote sequence (AD) were investigated by this rheological PIEZ system. After the addition of different NU172 concentrations, the coagulation of fresh human blood was analyzed under static conditions and using an in vitro rotation model under dynamic conditions (simulating ECC). The clotting times (CTs) detected by PIEZ were compared to those obtained with a medical reference device, a ball coagulometer. Additionally, after the circulation of blood samples for 30 min at 37 °C, blood cell numbers, thrombin markers (thrombin-antithrombin III (TAT) and fibrinopeptide A (FPA)) and a platelet activation marker (ß-thromboglobulin (ß-TG)) were analyzed by enzyme-linked immunosorbent assays (ELISAs). The increase of NU172 concentration resulted in prolonged CTs, which were comparable between the reference ball coagulometer and the PIEZ, demonstrating the reliability of the new measuring system. Moreover, by looking at the slope of the linear regression of the viscous and elastic components, PIEZ also could provide information on the kinetics of the coagulation reaction. The shear viscosity at the end of the measurements (after 300 s) was indicative of clot firmness. Furthermore, the PIEZ was able to detect the abrogation of coagulation inhibition after the equimolar addition of NU172 aptamer´s AD. The obtained results showed that the established PIEZ is capable to dynamically measure the hemostasis status in whole blood and can be applied to analyze nucleic acid-based drugs influencing the coagulation.

Impact of substrate elasticity on human hematopoietic stem and progenitor cell adhesion and motility.

In the bone marrow, hematopoietic stem cells (HSCs) reside in endosteal and vascular niches. The interactions with the niches are essential for the maintenance of HSC number and properties. Although the molecular nature of these interactions is well understood, little is known about the role of physical parameters such as matrix elasticity. Osteoblasts, the major cellular component of the endosteal HSC niche, flatten during HSC mobilization. We show that this process is accompanied by osteoblast stiffening, demonstrating that not only biochemical signals but also mechanical properties of the niche are modulated. HSCs react to stiffer substrates with increased cell adhesion and migration, which could facilitate the exit of HSCs from the niche. These results indicate that matrix elasticity is an important factor in regulating the retention of HSCs in the endosteal niche and should be considered in attempts to propagate HSCs in vitro for clinical applications.

Eligibility of a novel BW + technology and comparison of sensitivity and specificity of different imaging methods for radiological caries detection.

OBJECTIVES: Bitewing radiography is considered to be of high diagnostic value in caries detection, but owing to projections, lesions may remain undetected. The novel bitewing plus (BW +) technology enables scrolling through radiographs in different directions and angles. The present study aimed at comparing BW + with other 2D and 3D imaging methods in terms of sensitivity, specificity, and user reliability. MATERIALS AND METHODS: Five human cadavers were used in this study. In three cadavers, natural teeth were transplanted post-mortem. BW + , two-dimensional (digital sensors, imaging plates, 2D and 3D bitewing radiographs) and 3D methods (high and low dose CBCT) were taken. Carious lesions were evaluated on 96 teeth at three positions (mesial, distal, and occlusal) and scored according to their level of demineralization by ten observers, resulting in 35,799 possible lesions across all observers and settings. For reference, µCT scans of all teeth were performed. RESULTS: Overall, radiographic evaluations showed a high rate of false-negative diagnoses, with around 70% of lesions remaining undetected, especially enamel lesions. BW + showed the highest sensitivity for dentinal caries and had comparatively high specificity overall. CONCLUSIONS: Within the limits of the study, BW + showed great potential for added diagnostic value, especially for dentinal caries. However, the tradeoff of diagnostic benefit and radiation exposure must be considered according to each patient's age and risk.

Microstructural volumetric analysis of vertical alveolar ridge augmentation using autogenous tooth roots.

BACKGROUND: To volumetrically assess the bone microstructure following vertical alveolar ridge augmentation using differently conditioned autogenous tooth roots (TR) and second-stage implant placement. MATERIALS AND METHODS: The upper premolars were bilaterally extracted in n = 4 beagle dogs and randomly assigned to either autoclavation (TR-A) or no additional treatment (TR-C). Subsequently, TR were used as block grafts for vertical alveolar ridge augmentation in both lower quadrants. At 12 weeks, titanium implants were inserted and left to heal 3 weeks. Microcomputed tomography was used to quantify bone volume per tissue volume (BV/TV), trabecular thickness (Tb.Th), and trabecular spacing (Tb.Sp) at vestibular (v) and oral (o) aspects along the implant and in the augmented upper half of the implant, respectively. RESULTS: Median BV/TV [TR-C: 51.33% (v) and 70.42% (o) vs TR-A: 44.05% (v) and 64.46% (o)], Tb.th [TR-C: 0.22 mm (v) and 0.27 mm (o) vs TR-A: 0.23 mm (v) and 0.29 mm (o)] and Tb.Sp [TR-C: 0.26 mm (v) and 0.13 mm (o) vs TR-A: 0.29 µm (v) and 0.15 mm (o)] values were comparable in both groups. CONCLUSION: Both TR-C and TR-A grafts were associated with a comparable bone microstructure within the grafted area.

TNF-α modulation via Etanercept restores bone regeneration of atrophic non-unions.

Treatment of atrophic non-unions, especially in long bones is a challenging problem in orthopedic surgery due to the high revision and failure rate after surgical intervention. Subsequently, there is a certain need for a supportive treatment option besides surgical treatment. In our previous study we gained first insights into the dynamic processes of atrophic non-union formation and observed a prolonged inflammatory reaction with upregulated TNF-α levels and bone resorption. In this study we aimed to improve bone regeneration of atrophic non-unions via TNF-α modulation in a previously established murine femoral segmental defect model. Animals that developed atrophic non-unions of the femur after 5 and 10 weeks were treated systemically for 10 and 5 weeks with Etanercept, a soluble TNF-α antibody. µCT scans and histology revealed bony bridging of the fracture gap in the treatment group, while bone formation in control animals without treatment was not evident. Moreover, osteoclasts were markedly decreased via modulation of the RANKL/OPG axis due to Etanercept treatment. Additionally, immunomodulatory effects via Etanercept could be observed as further inflammatory agents, such as TGF-ß, IL6, MMP9 and 13 were decreased in both treatment groups. This study is the first showing beneficial effects of Etanercept treatment on bone regeneration of atrophic non-union formation. Moreover, the results of this study provide a new and promising therapeutic option which might reduce the failure rate of revision surgeries of atrophic non-unions.

Local Wnt3a treatment restores bone regeneration in large osseous defects after surgical debridement of osteomyelitis.

Impaired bone homeostasis caused by osteomyelitis provokes serious variations in the bone remodeling process, thereby involving multiple inflammatory cytokines to activate bone healing. We have previously established a mouse model for post-traumatic osteomyelitis and studied bone regeneration after sufficient debridement. Moreover, we could further characterize the postinfectious inflammatory state of bony defects after debridement with elevated osteoclasts and decreased bone formation despite the absence of bacteria. In this study, we investigated the positive effects of Wnt-pathway modulation on bone regeneration in our previous established mouse model. This was achieved by local application of Wnt3a, a recombinant activator of the canonical Wnt-pathway. Application of Wnt3a could enhance new bone formation, which was verified by histological and µ-CT analysis. Moreover, histology and western blots revealed enhanced osteoblastogenesis and downregulated osteoclasts in a RANKL-dependent manner. Further analysis of Wnt-pathway showed downregulation after bone infections were reconstituted by application of Wnt3a. Interestingly, Wnt-inhibitory proteins Dickkopf 1 (DKK1), sclerostin, and secreted frizzled protein 1 (sFRP1) were upregulated simultaneously to Wnt-pathway activation, indicating a negative feedback for active form of Beta-catenin. In this study, we could demonstrate enhanced bone formation in defects caused by post-traumatic osteomyelitis after Wnt3a application. KEY MESSAGES: Osteomyelitis decreases bone regeneration Wnt3a restores bone healing after infection Canonical Wnt-pathway activation with negative feedback.

Adipose-Derived Stromal Cells Are Capable of Restoring Bone Regeneration After Post-Traumatic Osteomyelitis and Modulate B-Cell Response.

Bone infections are a frequent cause for large bony defects with a reduced healing capacity. In previous findings, we could already show diminished healing capacity after bone infections, despite the absence of the causing agent, Staphylococcus aureus. Moreover, these bony defects showed reduced osteoblastogenesis and increased osteoclastogenesis, meaning elevated bone resorption ongoing with an elevated B-cell activity. To overcome the negative effects of this postinfectious inflammatory state, we tried to use the regenerative capacity of mesenchymal stem cells derived from adipose tissue (adipose-derived stem cells [ASCs]) to improve bone regeneration and moreover were curious about immunomodulation of applicated stem cells in this setting. Therefore, we used our established murine animal model and applicated ASCs locally after sufficient debridement of infected bones. Bone regeneration and resorption as well as immunological markers were investigated via histology, immunohistochemistry, Western blot, and fluorescence-activated cell scanning (FACS) analysis and µ-computed tomography (CT) analysis. Interestingly, ASCs were able to restore bone healing via elevation of osteoblastogenesis and downregulation of osteoclasts. Surprisingly, stem cells showed an impact on the innate immune system, downregulating B-cell population. In summary, these data provide a fascinating new and innovative approach, supporting bone healing after bacterial infections and moreover gain insights into the complex ceremony of stem cell interaction in terms of bone infection and regeneration. Stem Cells Translational Medicine 2019;8:1084-1091.