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Saliva was proposed as a diagnostic tool for systemic diseases. Here we determined the correlation of bone turnover markers in saliva, bone turnover markers in serum and bone mineral density in postmenopausal osteoporotic and healthy women. Forty postmenopausal osteoporotic and 40 age-matched healthy non-osteoporotic females were recruited for this case–control study. Serum and stimulated saliva levels of osteocalcin, N-terminal propeptide of type I collagen, bone-specific alkaline phosphatase and cross-linked-C-telopeptide of type I collagen were determined. Bone mineral density of the lumbar spine, proximal femur, and total hip were obtained. We show that osteocalcin and cross-linked-C-telopeptide of type I collagen (CTX) reached detectable levels in saliva while N-terminal propeptide of type I collagen and alkaline phosphatase were close or below the detection limit. Serum levels of bone turnover markers were significantly higher than saliva levels. Correlation analysis revealed a strong correlation of serum osteocalcin and, to a lesser extent, also serum CTX values with bone mineral density in lumbar spine, femoral neck, or total hip, respectively. There was, however, no significant correlation of bone mineral density with the respective bone turnover markers in saliva. There was a trend that saliva osteocalcin correlates with femoral neck (p = 0.16) or total hip (p = 0.06). There was also no association between serum and saliva bone turnover markers. This study reveals that saliva cannot replace the withdrawal of serum to evaluate bone metabolism.

Short-chain fatty acids (SCFA), bacterial metabolites released from dental biofilm, are supposed to target the oral epithelium. There is, however, no consensus on how SCFA affect the oral epithelial cells. The objective of the present study was to systematically review the available in vitro evidence of the impact of SCFA on human oral epithelial cells in the context of periodontal disease. A comprehensive electronic search using five databases along with a grey literature search was performed. In vitro studies that evaluated the effects of SCFA on human oral epithelial cells were eligible for inclusion. Risk of bias was assessed by the University of Bristol’s tool for assessing risk of bias in cell culture studies. Certainty in cumulative evidence was evaluated using GRADE criteria (grading of recommendations assessment, development, and evaluation). Of 3591 records identified, 10 were eligible for inclusion. A meta-analysis was not possible due to the heterogeneity between the studies. The risk of bias across the studies was considered “serious” due to the presence of methodological biases. Despite these limitations, this review showed that SCFA negatively affect the viability of oral epithelial cells by activating a series of cellular events that includes apoptosis, autophagy, and pyroptosis. SCFA impair the integrity and presumably the transmigration of leucocytes through the epithelial layer by changing junctional and adhesion protein expression, respectively. SCFA also affect the expression of chemokines and cytokines in oral epithelial cells. Future research needs to identify the underlying signaling cascades and to translate the in vitro findings into preclinical models.

OBJECTIVES The aim of this systematic review and meta-analysis was to compare the use of platelet-rich fibrin (PRF) with other commonly utilized treatment modalities for root coverage procedures. MATERIALS AND METHODS The eligibility criteria comprised randomized controlled trials (RCTs) comparing the performance of PRF with that of other modalities in the treatment of Miller class I or II (Cairo RT I) gingival recessions. Studies were classified into 5 categories as follows: (1) coronally advanced flap (CAF) alone vs CAF/PRF, (2) CAF/connective tissue graft (CAF/CTG) vs CAF/PRF, (3) CAF/enamel matrix derivative (CAF/EMD) vs CAF/PRF, (4) CAF/amnion membrane (CAF/AM) vs CAF/PRF, and (5) CAF/CTG vs CAF/CTG/PRF. Studies were evaluated for percentage of relative root coverage (rRC; primary outcome), clinical attachment level (CAL), keratinized mucosa width (KMW), and probing depth (PD) (secondary outcomes). RESULTS From 976 articles identified, 17 RCTs were included. The use of PRF statistically significantly increased rRC and CAL compared with CAF alone. No change in KMW or reduction in PD was reported. Compared with PRF, CTG resulted in statistically significantly better KMW and RC. No statistically significant differences were reported between the CAF/PRF and CAF/EMD groups or between the CAF/PRF and CAF/AM groups for any of the investigated parameters. CONCLUSIONS The use of CAF/PRF improved rRC and CAL compared with the use of CAF alone. While similar outcomes were observed between CAF/PRF and CAF/CTG for CAL and PD change, the latter group led to statistically significantly better outcomes in terms of rRC and KTW. In summary, the use of PRF in conjunction with CAF may represent a valid treatment modality for gingival recessions exhibiting adequate baseline KMW. CLINICAL RELEVANCE The data indicate that the use of PRF in conjunction with CAF statistically significantly improves rRC when compared with CAF alone but did not improve KMW. Therefore, in cases with limited baseline KMW, the use of CTG may be preferred over PRF.

This study aimed to compare the accuracy and reliability of digital versus conventional clinical measurements of the width of keratinized tissue. To this end, the keratinized tissue width was measured at 110 tooth sites in 5 pig jaws. The measurements were made at each site using three-dimensional (3D) scanned images and a periodontal probe. The actual keratinized tissue width was subsequently measured on histologic slides prepared from the same sites, and differences between the histologic slides and the digital and clinical measurements were analyzed to determine their accuracy in two measurement rounds. Furthermore, intrarater and interrater reliabilities were evaluated using the intraclass correlation coefficient (ICC). Here we show that the mean differences (and lower/upper limits of agreement) between the histologic and the digital/clinical measurements were 0.10 mm (−1.34/1.54 mm) and 1.11 mm (−0.69/2.92 mm), respectively, in the first round of measurements (p p p

Objectives Completion of adolescent growth represents the earliest time point for implant placement, yet craniofacial growth persists into adulthood and may affect implant position. We aimed to assess whether implants placed in the anterior maxillae of adults show angular changes over time. Material and methods We conducted a cephalometric pilot study in postpubertal patients with no growth disorders, skeletal malformations, or parafunctions. The patients received a single implant in the anterior maxilla and no orthodontic or orthognathic treatment afterwards. We measured angular changes of implants and central incisors on cephalograms taken immediately and after at least 5 years postoperatively with the Sella-Nasion line (SNL) and the nasal line (NL) as references. Changes in implant-SNL angles were the primary outcome. Results In 21 patients (30.2 ± 11.5 years at surgery) after a mean follow-up of 8.6 ± 1.3 years, implant-SNL angles and implant-NL angles changed in 81% and 57% of implants, respectively. Implant-SNL changes ranged from 3° counterclockwise to 4° clockwise and were more prevalent in males (100% vs. 58%) and patients under 30 at surgery (85% vs. 63%); mean absolute differences were larger in males (1.8 ± 1.0° vs. 1.3 ± 1.4°) and patients under 30 at surgery (1.5 ± 1.4° vs. 1.1 ± 1.4°). Incisor-SNL angles and incisor-NL angles changed in 89% and 32% of incisors, respectively. Conclusions Implants placed in the anterior maxillae of adults show modest angular changes over time. Clinical relevance Changes in implant angles have potential functional and esthetic consequences.

During the coronavirus disease 2019 (COVID-19) pandemic, aerosol-generating procedures, including dental implant treatments, are considered high-risk. With dental implant treatment mostly an elective procedure, we aimed to assess whether the pandemic influenced patient selection, surgical procedures, and postoperative complications. We compared dental implant treatments during (March to December 2020) and before (December 2018 to February 2020) the COVID-19 pandemic based on patient and implant parameters, as well as postoperative complications. For analysis, we used the Chi-squared test with the Holm–Sidak correction for multiple comparisons. The number of implants placed during the COVID-19 pandemic (696 implants in 406 patients, 70 implants per month) was comparable to pre-pandemic levels (1204 implants in 616 patients, 80 implants per month). Regarding patient parameters, there were no significant differences in respiratory (p = 0.69) and cardiovascular conditions (p = 0.06), diabetes (p = 0.69), and smoking (p = 0.68). Regarding implant parameters, there was a significant difference in the distribution of augmentative procedures (no augmentation, guided bone regeneration, and sinus floor elevation, p = 0.01), but no significant differences in the types of edentulous spaces (p = 0.19) and the timing of implant placement (p = 0.52). Regarding complications, there were significantly fewer minor complications (p < 0.001) and early (i.e., before loading) implant failures (p = 0.02) compared with pre-pandemic levels. Our results suggest that the COVID-19 pandemic had no effect on patient selection and only a slight effect on the surgical procedures. However, postoperative complications, including early failures, were significantly less prevalent during the pandemic.

Objectives Matrix metalloproteinases (MMPs) in dentin and saliva can degrade collagen. Divalent metals are known inhibitors of MMPs, but stannous – such as in the form of stannous chloride (SnCl2) or stannous fluoride (SnF2) – is yet to be tested for a possible inhibitory effect. In this study, we tested the inhibitory effect on the proteolytic activity of MMP-2 and MMP-9. Methods Sodium chloride (NaCl), sodium fluoride (NaF), and chlorhexidine (CHX) were used as controls. Gelatin zymography was performed with recombinant human MMP-2 and MMP-9. SnCl2, SnF2, NaF, NaCl, and CHX were included either in the incubation buffer (M1) or added to the recombinant MMPs (M2) before the MMPs were analyzed using zymography. Furthermore, the effect of SnCl2, SnF2, and NaF on the enzymatic activity of MMP-2 and MMP-9 was measured in human dentin either before or after acid etching using 37%phosphoric acid. The effect of SnCl2, NaF, and CHX on the viability and of SnCl2 and NaF on the proliferation of human gingival fibroblasts and L929 mouse fibroblasts was also determined. Results For M1, inhibitory concentrations (w/v%) of SnCl2 0.5% and 0.5%, SnF2 0.25% and 0.12%, NaF 0.12% and 0.5%, CHX 0.012% and 0.05%, were observed for MMP-2 and MMP-9, respectively. NaCl had no inhibitory effect. For M2, SnCl2 0.007% and 0.12%, and SnF2 0.03% and 0.5%, inhibited MMP-2 and MMP-9, respectively. NaF, NaCl and CHX had no effect. The enzymatic activity was slightly reduced when SnCl2 and NaF were applied on dentin before the acid attack. Regarding cell viability and proliferation of the cells after stimulation with the respective substances, NaF showed almost no effect, SnCl2 appeared to increase viability and proliferation of the cells, and CHX decreased the viability of cells. Conclusions Stannous ions caused a direct inhibition of the matrix metalloproteinases, whereas F− only had an inhibitory effect when added to the zymography buffer. Clinical significance Inhibition of MMPs using SnCl2 and SnF2 could play an important role in the prevention of dental erosion and caries. However, the clinical relevance of these findings needs to be proven.

Objective Hypertension is not only associated with cardiovascular diseases but also with alterations in bone quality. Hypertension therefore might be a risk factor for osseointegration. Preclinical studies suggest that losartan, an angiotensin II receptor blocker widely used to treat hypertension, has a beneficial effect in graft consolidation. Here, we determine the effect of hypertension and losartan on osseointegration. Methods We used spontaneously hypertensive rats (SHR) and normotensive Wistar albinus rats receiving losartan (30 mg/kg, p.o.) or left untreated. After 1 week, titanium miniscrews were inserted into the tibia. Sixty days after implantation, implant stability was evaluated by removal torque measurement considered the primary endpoint. Microcomputed tomography and histomorphometric analysis were secondary endpoints. Results Losartan increased the removal torque in the hypertensive SHR group to levels of the Wistar controls. While the cortical parameters of osseointegration remained unchanged, losartan increased medullary bone formation. Microcomputed tomography revealed a higher bone volume per tissue volume and trabecular thickness in the SHR rats treated with losartan. Histomorphometric analysis further showed that losartan significantly increased the thickness of newly formed bone in medullary area in hypertensive SHR rats. Losartan did not significantly alter the parameters of osseointegration in normotensive rats. Conclusions The data presented suggest that the angiotensin II receptor antagonist losartan increases the medullary parameters of osseointegration in a tibia model of spontaneously hypertensive rats. Considering the study limitations, understanding the impact of hypertension and the respective drugs on osseointegration requires further research.

OBJECTIVE To assess the impact of platelet-rich fibrin (PRF) on implant dentistry. The primary focused question was as follows: What are the clinical, histological, and radiographic outcomes of PRF administration for bone regeneration and implant therapy? METHOD A systematic literature search comprised three databases: MEDLINE, EMBASE, and Cochrane followed by a hand search of relevant scientific journals. Human studies using PRF for bone regeneration and implant therapy were considered and articles published up to December 31, 2017 were included. Eligible studies were selected based on the inclusion criteria. Randomized controlled trials (RCT) and controlled clinical trials (CCT) were included. RESULTS In total, 5,963 titles were identified with the search terms and by hand search. A total of 12 randomized controlled trials (RCT) met the inclusion criteria and were chosen for data extraction. Included studies focused on alveolar ridge preservation after tooth extraction, osseointegration process, soft tissue management, bone augmentation, bone regeneration after sinus floor elevation and surgical peri-implantitis treatment. Overall, the risk of bias was moderate or unclear. Nine studies showed superior outcomes for PRF for any of the evaluated variables, such as ridge dimension, bone regeneration, osseointegration process, soft tissue healing. Three studies failed to show any beneficial effects of PRF. No meta-analysis could be performed due to the heterogeneity of study designs. CONCLUSIONS There is moderate evidence supporting the clinical benefit of PRF on ridge preservation and in the early phase of osseointegration. It remains unclear whether PRF can reduce pain and improve soft tissue healing. More research support is necessary to comment on the role of PRF to improve other implant therapy outcomes.

OBJECTIVES: The task of this working group was to update the knowledge about the use of drugs and biologicals affecting healing of soft tissue and bone during implant treatment or procedures associated with it. Moreover, the impact of titanium particles and biocorrosion on complications and implant survival has been analysed. MATERIALS AND METHODS: The literature in the areas of interest (platelet concentrates, antiresorptive drugs as well as implant-host interaction) was screened using systematic reviews for the former two areas, whereas a narrative critical review was performed for the latter topic. Two manuscripts on platelet concentrates, one manuscript on antiresorptive drugs and one manuscript on the effects of biocorrosion, were presented for group analysis with subsequent discussion in the plenum and final consensus approval. RESULTS: Results and conclusions of the individual reviews of the three topics are presented in the respective papers. Conclusions of the group on strengths and weaknesses of available evidence as well as consensus statements and directions for further research are provided in this study. The following papers were subject to group discussions and formed the basis for the consensus statements: Stähli A, Strauss FJ, Gruber R. () The use of platelet-rich-plasma to enhance the outcomes of implant-related therapies: a systematic review Strauss FJ, Stähli A, Gruber R. (2018) The use of platelet-rich-fibrin to enhance the outcomes of implant-related therapies: a systematic review Mombelli A, Hashim D, Cionca N. () What is the impact of titanium particles and bio-corrosion on implant survival and complications? A critical review Stavropoulos A, Bertl K, Pietschmann P, Pandis N, Morten Schiødt, Klinge B. () The effect of antiresorptive drugs on implant therapy: a systematic review. ispartof: CLINICAL ORAL IMPLANTS RESEARCH vol:29 pages:93-99 ispartof: location:SWITZERLAND, Pfaffikon status: published

PURPOSE To evaluate the sinus membrane perforation and implant survival rate after crestal minimally invasive sinus floor augmentation using hydraulic pressure and vibrations. MATERIALS AND METHODS In this retrospective single cohort study, all patients who underwent minimally invasive sinus floor augmentation between 2007 and 2015 using hydraulic pressure and vibrations were included. The sinus membrane is elevated by physiologic saline at 1.5 bar. The fluid is then set into vibration to further separate the sinus membrane from the bony floor. The endpoints were sinus membrane perforation and the survival rate of implants. RESULTS The hydraulic pressure and vibration technique was applied in 156 patients. Seven patients with perforations of the sinus membrane were treated with the lateral window approach and excluded from the follow-up analysis. In the remaining 149 patients, 184 crestal sinus floor augmentations were performed and 184 implants were placed. In 10 of these 184 cases, a perforation was suspected in the postoperative computed tomography (CT) scan. In total, the perforation rate was 8.9% (17/191). Nineteen implants were lost during the follow-up period ranging from 0.2 to 8.4 years with a median of 2.3 years. The cumulative implant survival rates after 1, 3, and 5 years were 94.4%, 87.7%, and 87.7%, respectively. No severe perioperative complications were noted. CONCLUSION The hydraulic pressure and vibration technique allows a minimally invasive crestal sinus augmentation with a perforation rate less than 10% and implant survival rates of approximately 90%.

Bones with different embryological origin and mode of ossification are supposed to vary in their capacity for supporting graft consolidation. The aim of the current pilot study was to assess the TGF-β1 activity of bone chips obtained from distinct anatomic locations. Conditioned medium was prepared from bone chips harvested from pig calvaria, mandible, and tibia. Human oral fibroblasts were exposed to bone-conditioned medium (BCM) followed by reverse transcriptase polymerase chain reaction of the TGF-β1 target genes. Also an immunoassay for interleukin 11 (IL-11) and TGF-β1 was performed. The impact of BCM on alkaline phosphatase activity was determined with murine MC3T3-E1 osteogenic cells. The authors report here that BCM contains TGF-β1 in the ng/mL range. Bone chips prepared from pig calvaria, mandible, and tibia femur had a similar capacity for increasing the expression of the TGF-β1 target genes IL-11, NOX4, and PRG4. Correspondingly, immunoassays revealed similar production of IL-11 by human oral fibroblasts. Furthermore, conditioned medium obtained from the 3 bones decreased alkaline phosphatase activity in MC3T3-E1 osteogenic cells. These preliminary data demonstrate that particulated bone grafts, regardless of embryological origin, mode of ossification and morphology, release a similar TGF-β1 activity.

Background Hypertension is considered a risk factor in implant dentistry but the underlying reasons remain unclear. It may be that hypertension has a negative impact on the local bone quality. Purpose Here we evaluated the structural and histological parameters of bone collected from hypertensive patients treated by antagonists of the renin-angiotensin system, and of bone collected from normotensive patients. Material and methods A total of 30 patients were referred for rehabilitation with dental implants to be placed in the posterior mandible. The first group were hypertensive patients treated with RAS antagonists. The second group were normotensive patients not taking medication. Bone biopsies collected during implant installation were subjected to analysis. Micro CT revealed the structural parameters. Histological analyses together with immunohistochemical staining of osteogenic markers were performed. Results The structural parameters of bone volume, trabecular thickness, trabecular number, separation of the trabecular, and total porosity were similar between the 2 groups (P > .05). The histological appearance of bone derived from hypertensive patients was normal. The staining pattern of Runx-2, osteopontin, and osteocalcin were comparable in both groups. Conclusions These observations suggest that hypertensive patients treated with renin-angiotensin system antagonists have regular alveolar bone with respect to bone structure and histological parameters.

Purpose: Recombinant human bone morphogenetic protein 2 (rhBMP-2) together with an absorbable collagen carrier (ACS) was approved for augmentation of the maxillary sinus prior to implant placement. The original registration trial was based on a lateral window approach. Clinical outcomes of crestal sinus augmentation with rhBMP-2 have not been reported so far. Materials and Methods: An uncontrolled pilot trial in which seven patients with a residual maxillary height below 5 mm were enrolled to receive crestal sinus augmentation with rhBMP-2/ACS was conducted. Elevation of the sinus mucosa was performed by gel pressure. Primary endpoints were the gain in augmentation height and volume measured by computed tomography after 6 months. Evaluation of bone quality at the time of implant placement was based on histology. Secondary endpoints were the clinical and radiologic evaluation of the implants and patient satisfaction by visual analog scale (VAS) at the 2-year follow-up. Results: Median gain in augmentation height was 7.2 mm (range 0.0 to 17.5 mm). Five patients gained at least 5 mm of bone height. Two patients with a perforation of the sinus mucosa failed to respond to rhBMP-2/ACS and underwent lateral window augmentation. The median gain in augmentation volume of the five patients was 781.3 mm3 (range 426.9 to 1,242.8 mm3). Biopsy specimens showed a cancellous network consisting of primary plexiform bone with little secondary lamellar bone. After 2 years, implants were in function with no signs of inflammation or peri-implant bone loss. Patients were satisfied with the esthetic outcomes and chewing function. Conclusion: This pilot clinical trial supports the original concept that rhBMP-2/ACS supports bone formation, also in crestal sinus augmentation, and emphasizes the relevance of the integrity of the sinus mucosa to predict the bone gain.

Angiogenesis is essential for the consolidation of bone allografts. The underlying molecular mechanism, however, remains unclear. Soluble factors released from demineralized freeze-dried bone target mesenchymal cells; however, their effect on endothelial cells has not been investigated so far. The aim of the present study was therefore to examine the effect of conditioned medium from demineralized freeze-dried bone on human umbilical endothelial cells in vitro. Conditioned medium was first prepared from demineralized freeze-dried bone following 24 hours incubation at room temperature to produce demineralized bone conditioned media. Thereafter, conditioned medium was used to stimulate human umbilical vein endothelial cells in vitro by determining the cell response based on viability, proliferation, expression of apoptotic genes, a Boyden chamber to determine cell migration, and the formation of branches. The authors report here that conditioned medium decreased viability and proliferation of endothelial cells. Neither of the apoptotic marker genes was significantly altered when endothelial cells were exposed to conditioned medium. The Boyden chamber revealed that endothelial cells migrate toward conditioned medium. Moreover, conditioned medium moderately stimulated the formation of branches. These findings support the concept that conditioned medium from demineralized freeze-dried bone targets endothelial cells by decreasing their proliferation and enhancing their motility under these in vitro conditions.

Background Inflammatory osteolysis is the clinical hallmark of peri-implantitis. The morphology of the remaining peri-implant bone and the level of osseointegration, however, remain unknown. Our aim was to characterize advanced peri-implantitis bone defects in humans. Methods Four patients (3 female and 1 male) were diagnosed with peri-implantitis. A total of 5 implants with machined surfaces and a mean loading time of 12 ± 6 years were removed due to advanced bone loss. The defect extension, the peri-implant bone density (bone area per tissue area in percentage), bone-to-implant contact (%), and the number of filled and empty osteocyte lacunae were calculated based on undecalcified histological specimens. Results The defect extension was on average 4.2 mm (95% CI 0.8–3.4). Remaining peri-implant bone showed a high density of 85.5% (95% CI 79.1–91.3) and covered in total 74% (95% CI 70.5–77.5) of the implant surface. Filled and empty osteocyte lacunae density was on average 191 and 165/mm2 (95% CI 132–251; 103–225), respectively. Histology further revealed signs of ongoing bone formation and resorption. Conclusion There are signs that suggest that once the original cortical bone is lost due to peri-implantitis, the remaining apical trabecular bone is reinforced and transformed into cortical bone that might take over the functional load.

Solid platelet-rich fibrin (PRF) is produced with centrifugation tubes designed to accelerate clotting. Thus, activated platelets may accumulate within the fibrin-rich extracellular matrix even before centrifugation is initiated. It can thus be assumed that platelets and their growth factors such as transforming growth factor-β (TGF-β) are trapped within PRF independent of their relative centrifugal force (RCF), the gravitation or g-force. To test this assumption, we prepared PRF membranes with tubes where clotting is activated by a silicone-coated interior. Tubes underwent 210 g, 650 g and 1500 g for 12 min in a horizontal centrifuge. The respective PRF membranes, either in total or separated into a platelet-poor plasma and buffy coat fraction, were subjected to repeated freeze-thawing to prepare lysates. Gingival fibroblasts were exposed to the PRF lysates to provoke the expression of TGF-β target genes. We show here that the expression of interleukin 11 (IL11) and NADPH oxidase 4 (NOX4), and Smad2/3 signaling were similarly activated by all lysates when normalized to the size of the PRF membranes. Notably, platelet-poor plasma had significantly less TGF-β activity than the buffy coat fraction at both high-speed protocols. In contrast to our original assumption, the TGF-β activity in PRF lysates produced using horizontal centrifugation follows a gradient with increasing concentration from the platelet-poor plasma towards the buffy coat layer.

Deproteinized bovine bone mineral (DBBM) is brittle and can break into fragments. Here, we examined whether DBBM fragments have an impact on mice calvarial bone during bone augmentation. DBBM was either randomly crushed (DBBM fragments) or left undisturbed (DBBM granules). Then, DBBM fragments or original DBBM granules were placed onto calvarial bone in 20 BALB/c mice. Following random allocation, ten mice received DBBM fragments and ten mice received original DBBM granules. After fourteen days of healing, micro computed tomography (micro-CT) and histological analysis of the augmented sites were performed. The primary outcome was the porosity of the calvarial bone. The micro-CT analysis revealed that DBBM fragments failed to significantly change the porosity of the calvarial bone as compared with original DBBM granules, despite the slightly higher bone resorption in the DBBM fragment group, 10.3% (CI 6.3&ndash, 11.6) versus 6.1% (CI 4.1&ndash, 7.8, p = 0.355), respectively. The cortical bone volume was not altered by DBBM fragments as compared with original DBBM granules, i.e., 79.0% (CI 78.9&ndash, 81.2) versus 81.5% (CI 80.1&ndash, 83.3, p = 0.357), respectively. The DBBM fragment group revealed similar bone thickness values as compared with the DBBM granules group, i.e., 0.26 mm (CI 0.23&ndash, 0.29) versus 0.25 mm (CI 0.22&ndash, 0.27, p = 0.641), respectively. The histological evaluation supported the micro-CT observations, displaying minor signs of porosity and resorption. The particle-size distribution analysis confirmed a shift towards smaller particle sizes in the DBBM fragment group. These findings suggest that DBBM fragments behave similarly to original DBBM granules in terms of bone morphological changes at augmented sites.

Hypoallergenic formulas are recommended for infants who are not breastfed and cannot tolerate cow milk formulas due to allergy. These formulas are hydrolyzed to break down larger protein chains into shorter, easy-to-digest, and potentially less allergenic proteins. Hydrolysis, however, possibly occurs at the expense of the transforming growth factor beta (TGF-β) and anti-inflammatory activity that is inherent in regular formula. Our objective was to determine the TGF-β and the anti-inflammatory activity of commercially available hypoallergenic and regular formulas. Human gingival fibroblasts were incubated with reconstituted formulas followed by detection of TGF-β target genes and activation of Smad2/3 signaling. Gingival fibroblasts and the oral squamous cell carcinoma cell line HSC-2 were also exposed to formulas before adding interleukin (IL)1β and tumor necrosis factor (TNF)α to provoke expression of pro-inflammatory cytokines. For murine bone marrow-derived macrophages, pro-inflammatory cytokine expression was stimulated with saliva. Changes in p65 nuclear translocation and phosphorylation of smad3 and p38 were analyzed by immunostaining. Our study demonstrated that regular formula, but not hypoallergenic formula, enhanced the expression of TGF-β target genes IL11, PRG4, and NOX4 in gingival fibroblasts. Hypoallergenic formulas also failed to initiate nuclear translocation of Smad2/3 and phosphorylation of Smad3. Moreover, regular formulas were more potent than hypoallergenic formulas in reducing the expression of pro-inflammatory cytokines in gingival fibroblasts, HSC-2 epithelial cells, and murine bone marrow macrophages. Hypoallergenic and regular formulas had a similar capacity to reduce p65 nuclear translocation and phosphorylation of p38 in fibroblasts. These findings suggest that hypoallergenic formulas lack in vitro TGF-β activity and have a lower anti-inflammatory activity compared with regular formulas.

Objectives Risk prediction in implant dentistry presents specific challenges including the dependence of observations from patients with multiple implants and rare outcome events. The aim of this study was to use advanced statistical methods based on penalized regression to assess risk factors in implant dentistry. Material and methods We conducted a retrospective study from January 2016 to November 2018 recording postoperative complications including bleeding, hematoma, local infection, and nerve damage, as well as early implant failure. We further assessed patient‐ and implant‐related risk factors including smoking and diabetes, as well as treatment parameters including types of gaps and surgical procedures. Univariable and multivariable generalized estimating equation (GEE) models were estimated to assess predictor effects, and a prediction model was fitted using L1 penalized estimation (lasso). Results In a total of 1,132 patients (mean age: 50.6 ± 16.5 years, 55.4% female) and 2,413 implants, postoperative complications occurred in 71 patients. Sixteen implants were lost prior to loading. Multivariable GEE models showed a higher risk of any complication for diabetes mellitus (p = .006) and bone augmentation (p = .039). The models further revealed a higher risk of local infection for bone augmentation (p = .003), and a higher risk of hematoma formation for diabetes mellitus (p = .007) and edentulous jaws (p = .024). The lasso model did not select any risk factors into the prediction model. Conclusions Using novel methodology well‐suited to tackle the specific challenges of risk prediction in implant dentistry, we were able to reliably estimate associations of risk factors with outcomes.

OBJECTIVES Emdogain, containing an extract of fetal porcine enamel matrix proteins, is a potent stimulator of in vitro osteoclastogenesis. The underlying molecular mechanisms are, however, unclear. MATERIAL AND METHODS Here, we have addressed the role of transforming growth factor-beta receptor type 1 (TGF-βRI) kinase activity on osteoclastogenesis in murine bone marrow cultures. RESULTS Inhibition of TGF-βRI kinase activity with SB431542 abolished the effect of Emdogain on osteoclastogenesis induced by receptor activator of nuclear factor kappa-B ligand or tumor necrosis factor-alpha. SB431542 also suppressed the Emdogain-mediated increase of OSCAR, a co-stimulatory protein, and dendritic cell-specific transmembrane protein and Atp6v0d2, the latter two being involved in cell fusion. Similar to transforming growth factor-beta1 (TGF-β), Emdogain could not compensate for the inhibition of IL-4 and IFNγ on osteoclast formation. When using the murine macrophage cell line RAW246.7, SB431542 and the smad-3 inhibitor SIS3 blocked Emdogain-stimulated expression of the transcription factor NFATc1. CONCLUSIONS Taken together, the data suggest that TGF-βRI kinase activity is necessary to mediate in vitro effects of Emdogain on osteoclastogenesis. CLINICAL RELEVANCE Based on these in vitro data, we can speculate that at least part of the clinical effects of Emdogain on osteoclastogenesis is mediated via TGF-β signaling.

AIMS The objective of this study is to evaluate the effects of a paste-like bone substitute material with easy handling properties and improved mechanical stability on periodontal regeneration of intrabony defects in dogs. MATERIALS AND METHODS Mandibular and maxillary first and third premolars were extracted, and three-wall intrabony defects were created on second and fourth premolars. After a healing period of 3 months, acute type defects were filled with a paste-like formulation of deproteinized bovine bone mineral (DBBM) (particle size, 0.125-0.25 mm) in a collagenous carrier matrix (T1), pulverized DBBM (particle size, 0.125-0.25 mm) without the carrier (T2), or Bio-Oss® granules (particle size, 0.25-1.00 mm) as control (C). All defects were covered with a Bio-Gide® membrane. The dogs were sacrificed after 12 weeks, and the specimens were analyzed histologically and histometrically. RESULTS Postoperative healing of all defects was uneventful, and no histological signs of inflammation were observed in the augmented and gingival regions. New cementum, new periodontal ligament, and new bone were observed in all three groups. The mean vertical bone gain was 3.26 mm (T1), 3.60 mm (T2), and 3.81 mm (C). That of new cementum was 2.25 mm (T1), 3.88 mm (T2), and 3.53 mm (C). The differences did not reach statistical significance. The DBBM particles were both incorporated in new bone and embedded in immature bone marrow. CONCLUSIONS The results of this preclinical study showed that the 0.125-0.25-mm DBBM particles in a powder or paste formulation resulted in periodontal regeneration comparable to the commercially available DBBM. Osteoconductivity, in particular, was not affected by DBBM size or paste formulation. CLINICAL RELEVANCE The improved handling properties of the paste-like bone substitute consisting of small DBBM particles embedded in a collagen-based carrier hold promise for clinical applications.

Objective This study investigated the influence of the degree of conversion (DC), resin-based composites (RBC) composition, and the effect of additional violet light from one light curing unit (LCU) on cell attachment/growth, eluate cytotoxicity, and gene expression. Methods The effect of different DC of RBCs on human gingival fibroblasts (HGFs) when cultured directly onto cured RBCs, and when exposed afterwards to eluates in cell culture medium was examined. Venus® (RBC-V; Bis-GMA-based) and Venus Pearl® (RBC-P; TCD-DI-HEA and UDMA-based) were cured using a single emission peak (blue) light, Translux Wave®; TW and a dual emission peak (blue-violet) light, Translux 2 Wave®; T2W. To determine the value of the additional violet light from the T2W, exposure times and distances were adjusted to deliver similar radiant exposures (RE) from the blue region of both lights at five different RE levels from 1.5 J/cm² to 28.9 J/cm². Results Both RBCs light-cured with the T2W at higher REs resulted in higher DC, increased cell adhesion and decreased eluate cytotoxicity. RBC-V induced greater cell adhesion, lower mRNA levels of pro-inflammatory markers, and higher mRNA levels of a proliferation marker than RBC-P. Wettability was the same for both RBCs. Toxicity decreased with increasing number of elution cycles. The initial eluates from RBC-P had a lower toxicity than from RBC-V. Significance RBCs cured with T2W (delivering both blue and violet light) at higher RE had greater DCs. The greatest DC and the least cell reactions were observed when the RE was >25 J/cm².

Background Platelet‐rich fibrin (PRF) serves as a reservoir of bioactive molecules to support wound healing and bone regeneration. The beneficial action of PRF might involve macrophage polarization from proinflammatory M1 toward pro‐resolving M2 phenotypes. This study aims to evaluate the effect of PRF on macrophage polarization. Methods Murine primary macrophages and RAW 264.7 cells were exposed to saliva and lipopolysaccharides (LPS) with and without PRF lysates obtained by repeated freeze‐thawing or the secretome of PRF membranes, termed PRF conditioned medium. The expression of the M1 marker genes interleukin 1β (IL1β) and interleukin 6 (IL6) along with the M2 markers arginase‐1 and chitinase‐like 3 (Chil3 or YM1) were evaluated by real time polymerase chain reaction. Immunoassay and immunofluorescence staining were performed for IL6 and p65 translocation, a subunit nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐kB), respectively. Results We report here that PRF lysates and PRF conditioned medium, the latter containing the secretome, greatly decreased the proinflammatory response of primary macrophages and RAW 264.7 cells as indicated by the expression of IL1β and IL6. The anti‐inflammatory activity of PRF lysates was further confirmed by IL6 immunoassay. Moreover, PRF lysates suppressed the translocation of p65 from the cytoplasm into the nucleus after incubation with saliva. In support of M2 polarization, PRF lysates and PRF conditioned medium enhanced the expression of arginase‐1 and YM1 in primary macrophages. Conclusion Our results indicate that PRF holds an anti‐inflammatory activity and shifts the macrophage polarization from an M1 toward an M2 phenotype.

Background Platelet‐rich fibrin (PRF) membranes can preserve alveolar ridge dimension after tooth extraction. Thus, it can be presumed that PRF suppresses the catabolic events that are caused by osteoclastic bone resorption. Methods To address this possibility, we investigated the impact of soluble extracts of PRF membranes on in vitro osteoclastogenesis in murine bone marrow cultures. Osteoclastogenesis was induced by exposing murine bone marrow cultures to receptor activator of nuclear factor kappa B ligand (RANKL), macrophage colony‐stimulating factor (M‐CSF) and transforming growth factor‐beta 1 (TGF‐β1) in the presence or absence of PRF. Osteoclastogenesis was evaluated based on histochemical, gene expression, and resorption analysis. Viability was confirmed by formation of formazan crystals, live‐dead staining and caspase‐3 activity assay. Results We report here that in vitro osteoclastogenesis is greatly suppressed by soluble extracts of PRF membranes as indicated by tartrate‐resistant acid phosphatase (TRAP) staining and pit formation. In support of the histochemical observations, soluble extracts of PRF membranes decreased expression levels of the osteoclast marker genes TRAP, Cathepsin K, dendritic cell‐specific transmembrane protein (DCSTAMP), nuclear factor of activated T‐cells (NFATc1), and osteoclast‐associated receptor (OSCAR). PRF membranes, however, cannot reverse the process once osteoclastogenesis has evolved. Conclusion These in vitro findings indicate that PRF membranes can inhibit the formation of osteoclasts from hematopoietic progenitors in bone marrow cultures. Overall, our results imply that the favorable effects of PRF membranes in alveolar ridge preservation may be attributed, at least in part, by the inhibition of osteoclastogenesis.

Objectives Saliva can support oral wound healing, a process that requires a temporary inflammatory reaction. We have reported previously that saliva provokes a strong inflammatory response in oral fibroblasts. Bone marrow cells also give rise to macrophages, a heterogeneous subset of cell population involved in wound healing. Lipopolysaccharide (LPS) and interleukin 4 (IL-4) induce activation of pro-(M1), and anti-(M2) inflammatory macrophages, respectively. Yet, the impact of saliva on programming bone marrow cells into either M1 or M2 macrophages remains unclear . Design Herein, we examined whether sterile saliva affects the in vitro process of macrophage polarization based on murine bone marrow cultures and RAW264.7 mouse macrophages. Results We report that sterile saliva, similar to lipopolysaccharides, provoked a robust activation of the M1 phenotype which is characterized by a strong increase of the respective genes IL-12 and IL-6, based on a real-time gene expression analysis, and for IL-6 with immunoassay. Arginase-1 and Ym1, both genes characteristic for the M2 phenotype, were not considerably modulated by saliva. Inhibition of TLR4 signaling with TAK-242, blocking NFκB signaling with Bay 11-7085, but also autoclaving saliva greatly reduced the development of the M1 phenotype. Conclusion These data suggest that saliva activates the TLR4 dependent polarization into pro-inflammatory M1 macrophages in vitro.

Enamel matrix derivative (EMD) and collagen membranes (CMs) are simultaneously applied in regenerative periodontal surgery. The aim of this study is to evaluate the ability of two CMs and a collagen matrix to adsorb the activity intrinsic to EMD that provokes transforming growth factor (TGF)-β signaling in oral fibroblasts.Three commercially available collagen products were exposed to EMD or recombinant TGF-β1, followed by vigorous washing. Oral fibroblasts were either seeded directly onto collagen products or were incubated with the respective supernatant. Expression of TGF-β target genes interleukin (IL)-11 and proteoglycan 4 (PRG4) was evaluated by real time polymerase chain reaction. Proteomic analysis was used to study the fraction of EMD proteins binding to collagen.EMD or TGF-β1 provoked a significant increase of IL-11 and PRG4 expression of oral fibroblasts when seeded onto collagen products and when incubated with the respective supernatant. Gene expression was blocked by the TGF-β receptor I kinase inhibitor SB431542. Amelogenin bound most abundantly to gelatin-coated culture dishes. However, incubation of palatal fibroblasts with recombinant amelogenin did not alter expression of IL-11 and PRG4.These in vitro findings suggest that collagen products adsorb a TGF-β receptor I kinase-dependent activity of EMD and make it available for potential target cells.

Es sind nun mehr als 12 Jahre seit den ersten Publikationen uber die medikamentenassoziierte Osteonekrose des Kiefers (MRONJ) vergangen. Der Mangel an wissenschaftlich fundierter Information hat in dieser Zeit haufig zu Fehlinformationen und Angsten bei Patienten und auch bei Arzten/Zahnarzten gefuhrt. Eine MRONJ ist zweifelsfrei eine schwerwiegende Erkrankung der Mundhohle mit starker Beeintrachtigung der Lebensqualitat. Sie kann als Therapiefolge nach Behandlung von malignen und benignen Knochenerkrankungen mit Bisphosphonaten, Denosumab oder Bevacizumab (Angiogenesehemmer) auftreten. Die MRONJ ist eine seltene Erkrankung und tritt in erster Linie bei onkologischen Patienten (1–15 %) auf, die in kurzeren Abstanden und mit hohen Dosen behandelt werden. Hier uberwiegt der nachgewiesene Nutzen einer Behandlung mit antiresorptiv wirkenden Medikamenten meist bei Weitem die potenziellen Risiken der Entstehung einer MRONJ. Insbesondere onkologische Patienten sollten vor Behandlungsbeginn zahnarztlich versorgt sein. Bei einer Osteoporosetherapie tritt eine Osteonekrose des Kiefers als assoziierte Therapiefolge sehr selten auf (0,001–0,01 %) und ist bei einer Therapiedauer unter 4 Jahren kaum hoher als die Inzidenz in der Normalbevolkerung (< 0,001 %). Trotzdem ist bei Patienten mit einer diesbezuglichen Therapie wegen des potenziellen Risikos einer MRONJ eine erhohte Achtsamkeit geboten. Praventive Masnahmen sind sinnvoll, notwendig und wirksam. Nach 4 Jahren nimmt die Inzidenz um den Faktor 10 zu. Zur Therapie der manifesten MRONJ kann bei umschriebenen Befunden eine konservative Therapie oder eine wenig invasive Operation zielfuhrend sein. Bei fortgeschrittenen oder multiplen Kiefernekrosen ist aber oft auch eine operative Therapie unter strengen Vorsichtsmasnahmen medizinisch notwendig und gewinnt zunehmend an Bedeutung. Das individuelle Risikoprofil fur eine MRONJ ist somit bei jedem einzelnen Patienten unterschiedlich und muss dem Risiko der zu behandelnden Grunderkrankung gegenubergestellt werden. Die wirkungsvollste und einfachste Masnahme gegen eine MRONJ ist die Pravention. Um praventive Masnahmen optimal einzusetzen und auch um Therapieunsicherheiten zu vermeiden, ist eine enge Zusammenarbeit zwischen dem osteologisch/onkologisch behandelnden Arzt und dem behandelnden Zahnarzt unbedingt notwendig.

Periodontal disease is associated with chronic oxidative stress and inflammation. Caffeic acid phenethyl ester (CAPE), which is a potent inducer of heme oxygenase 1 (HO1), is a central active component of propolis, and the application of propolis improves periodontal status in diabetic patients. Here, primary murine macrophages were exposed to CAPE. Target gene expression was assessed by whole-genome microarray, RT-PCR and Western blotting. The antioxidative and anti-inflammatory activities of CAPE were examined by exposure of the cells to hydrogen peroxide, saliva and periodontal pathogens. The involvement of HO1 was investigated with the HO1 inhibitor tin protoporphyrin (SnPP) and knockout mice for Nrf2, which is a transcription factor for detoxifying enzymes. CAPE increased HO1 and other heat shock proteins in murine macrophages. A p38 MAPK inhibitor and Nrf2 knockout attenuated CAPE-induced HO1 expression in macrophages. CAPE exerted strong antioxidative activity. Additionally, CAPE reduced the inflammatory response to saliva and periodontal pathogens. Blocking HO1 decreased the antioxidative activity and attenuated the anti-inflammatory activity of CAPE. In conclusion, CAPE exerted its antioxidative effects through the Nrf2-mediated HO1 pathway and its anti-inflammatory effects through NF-κB inhibition. However, preclinical models evaluating the use of CAPE in periodontal inflammation are necessary in future studies., Healthy gums: a little help from honeybees Propolis, also known as ‘honeybee glue,’ may protect teeth and gums against periodontal disease. In periodontal disease, chronic inflammation and oxidative damage harm gum tissue and lead to tooth loss; propolis has been shown to improve periodontal health for patients with diabetes. Bees make propolis by mixing beeswax, honey, plant resins and their own saliva, and use it to patch honeycomb and prevent growth of microbes in the hive. Reinhard Gruber of the Department of Oral Biology at the Medical University of Vienna and of the Department of Periodontology, University of Bern and co-workers investigated the effects of one of propolis’ active ingredients, caffeic acid phenethyl ester (CAPE), on oxidative stress and inflammation. They found that CAPE reduced oxidative damage and dampened inflammation; further investigation revealed the genetic basis of the beneficial effects, paving the way for future clinical studies. These results may help identify alternative treatments for periodontal disease.

Objectives Osteoblasts lay down new bone on implant surfaces. The underlying cellular mechanism and the spatio‐temporal mode of action, however, remain unclear. It can be proposed that growth factors released upon acidification by osteoclasts adsorb to the implant surface and control the early stages of osseointegration. Methods To simulate bone lysis by osteoclasts, titanium discs were exposed to acid bone lysate (ABL) followed by vigorous washing and seeding of oral fibroblasts. The expression of TGF‐β target genes interleukin 11 (IL11) and NADPH oxidase 4 (NOX4) was evaluated by reverse transcriptase polymerase chain reaction and IL11 ELISA. TGF‐β signaling activation was assessed via Smad2/3 immunofluorescence. The impact of ABL on osteogenic differentiation was determined with murine ST2 mesenchymal stromal cells. Results We report here that ABL‐conditioned titanium discs, independent of turned or rough surface, increased the expression of IL11 and NOX4. This increase was blocked by the TGF‐β receptor 1 antagonist SB431542. Further support for the TGF‐β signaling activation came from the translocation of Smad2/3 into the nucleus of oral fibroblasts. Moreover, titanium discs exposed to ABL decreased alkaline phosphatase and osteopontin in ST2 cells. Conclusions These in vitro findings suggest that titanium can adsorb TGF‐β from ABLs. The data provide a strong impetus for studies on the protein adsorption on implant surfaces in vitro and in vivo, specifically for growth factors including bone‐derived TGF‐β during successful and failed osseointegration.

The incisors of rodents comprise an iron-rich enamel and grow throughout adult life, making them unique models of iron metabolism and tissue homeostasis during aging. Here, we deleted Atg7 (autophagy related 7) in murine ameloblasts, i.e. the epithelial cells that produce enamel. The absence of ATG7 blocked the transport of iron from ameloblasts into the maturing enamel, leading to a white instead of yellow surface of maxillary incisors. In aging mice, lack of ATG7 was associated with the growth of ectopic incisors inside severely deformed primordial incisors. These results suggest that 2 characteristic features of rodent incisors, i.e. deposition of iron on the enamel surface and stable growth during aging, depend on autophagic activity in ameloblasts. Abbreviations: ATG5: autophagy related 5; ATG7: autophagy related 7; CMV: cytomegalovirus; Cre: Cre recombinase; CT: computed tomography; FTH1: ferritin heavy polypeptide 1; GFP: green fluorescent protein; KRT5: keratin 5; KRT14: keratin 14; LGALS3: lectin, galactose binding, soluble 3; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; NCOA4: nuclear receptor coactivator 4; NRF2: nuclear factor, erythroid 2 like 2; SQSTM1: sequestosome 1.

Objectives Deproteinized bovine bone mineral (DBBM) is not resorbable. However, the behavior of DBBM under inflammatory conditions remains unclear. Aim of the study was therefore to evaluate the resorption of DBBM under local inflammatory conditions in vivo using the calvarial osteolysis model. Methods In thirty adult BALB/c mice, DBBM was implanted into the space between the elevated soft tissue and the calvarial bone. Inflammation was induced either by lipopolysaccharides (LPS) injection or by polyethylene particles (Ceridust) mixed with DBBM. Three modalities were randomly applied (n = 10 each): (a) DBBM alone (control), (b) DBBM + LPS, and (c) DBBM + polyethylene particles (Ceridust). Mice were euthanized on day fourteen, and each calvarium was subjected to histological and µCT analysis. Primary outcome was the size distribution of the DBBM particles. Secondary outcome was the surface erosion of the calvarial bone. Results Histological and µCT analysis revealed that the size distribution and the volume of DBBM particles in the augmented site were similar between DBBM alone and the combinations with LPS or polyethylene particles. Moreover, histological evaluation showed no signs of erosions of DBBM particles under inflammatory conditions. µCT analysis and histology further revealed that LPS and the polyethylene particles, but not the DBBM alone, caused severe erosions of the calvarial bone as indicated by large voids representing the massive compensatory new immature woven bone formation on the endosteal surface. Conclusions Local calvarial bone but not the DBBM particles undergo severe resorption and subsequent new bone formation under inflammatory conditions in a mouse model.

To describe the biology of alveolar bone regeneration.Four comprehensive reviews were performed on (a) mesenchymal cells and differentiation factors leading to bone formation; (b) the critical interplay between bone resorbing and formative cells; (c) the role of osteoimmunology in the formation and maintenance of alveolar bone; and (d) the self-regenerative capacity following bone injury or tooth extraction were prepared prior to the workshop.This summary information adds to the fuller understanding of the alveolar bone regenerative response with implications to reconstructive procedures for patient oral rehabilitation. The group collectively formulated and addressed critical questions based on each of the reviews in this consensus report to advance the field. The report concludes with identified areas of future research.

To evaluate the effects of bracket removal using an erbium laser on the pulp temperature and enamel surface.Removal of orthodontic brackets with conventional debonding pliers may result in enamel cracks. To avoid damage to the enamel surface and effectively remove metal or ceramic brackets, different types of lasers, such as Nd:YAG, COA total of 55 brackets (n = 55; 20 metal and 35 ceramic ones) were bonded to 55 caries-free premolars extracted for orthodontic indications. Brackets were irradiated with Er:YAG laser (Morita, Irvine, CA) with a wavelength of 2940 nm at a power of 3.4 W, energy 170 mJ, frequency 20 Hz, pulse duration 300 μs, tip diameter 0.8 mm, air/fluid cooling 3 mL/s, and time of irradiation: 6 sec. Debonding was made by scanning (n = 15; 6 sec irradiation at distance of 2 mm from the bracket with an "S" shape movement) and circular (n = 15; 6 sec irradiation at distance of 1 mm from the bracket) motion technique in ceramic brackets or the circular motion technique in metal brackets (n = 15). The number of 10 nonirradiated teeth with ceramic (n = 5) or metal brackets (n = 5) was used as a control in SEM test and EDS analysis. The damage in tooth enamel surface and the calcium percentage were analyzed by means of scanning electron microscope (JEOL 6610LV, JEOL, Japan) and energy dispersive X-ray spectroscopy (EDS, Oxford, United Kingdom). Temperature changes in the pulp were measured by K-type thermocouple. Evaluation of the Adhesive Remnant Index (ARI) on the enamel surface of each tooth was examined after bracket debonding.The scanning method has caused significantly lower temperature increase (mean: 0.83°C) compared with circular motion technique around the ceramic brackets (mean: 1.78°C; p = 0.0001) or the metal brackets (mean: 1.29°C; p = 0.015). ARI score showed no differences between the study groups (p = 0.57). SEM analysis revealed no cracks on enamel surface after laser-assisted debonding in comparison with the control samples where cracks were found. EDS showed a higher mean percentage of the calcium (30.7-85.8%) for all test groups compared with control samples (mean: 7%; p = 0.0002). The amount of the calcium elements was higher for metal brackets in comparison with ceramic ones (p = 0.0002).Er:YAG laser-assisted debonding causes a minor increase in the pulp temperature and reduced the risk of enamel damage compared with conventional bracket removal.

Extracorporeal irradiation sterilizes resected tumor bone used as autografts in reconstruction surgery. Therapeutic irradiation is a standard technique in head and neck cancer therapy that aims to preserve organ function. Bone irradiation has a complex, mostly inhibitory, effect on remodeling and regeneration, although the underlying mechanisms are still not fully understood. It remains unclear if extracorporeal irradiation affects the paracrine-like activity of the corresponding autografts. We recently reported that bone-conditioned medium from autogenous bone chips contains a number of factors that might affect cell activity. In the present study, we investigated the effects of extracorporeal irradiation of porcine cortical bone chips on the activity of the corresponding bone-conditioned medium. The effects of bone-conditioned medium on the expressions of transforming growth factor-beta (TGF-β) target genes in oral fibroblasts were assessed. Bone-conditioned medium from bone chips exposed to a total radiation dose up to 120 Gy did not affect expressions of TGF-β target genes, including adrenomedullin, BTB/POZ domain-containing protein 11, proteoglycan 4, NADPH oxidase 4, and interleukin 11, in oral fibroblasts. In conclusion, bone irradiation does not alter the capability of the corresponding bone-conditioned medium to provoke a robust fibroblastic cell response in vitro. (J Oral Sci 58, 325-331, 2016).

BACKGROUND Current strategies to reduce medical device-associated infections propose zirconia as a potential implant material which may limit bacterial adhesion. Because multinucleated giant cells (MNGCs) have been detected on these implant surfaces, concerns have been raised regarding tissue integration. PURPOSE The present study examined the presence of MNGCs and their subsequent effect upon tissue integration. Surface-modified implants made of yttria-stabilized (TZP) and alumina-toughened zirconia (ATZ) were compared with commercially pure titanium (Ti). MATERIALS AND METHODS Seven miniature pigs received three implants on either side of the maxilla. After healing periods of 4 and 8 weeks, the tissue response at the implant surfaces was characterized according to three specific parameters: bone-to-implant contact (BIC), MNGC-to-implant contact (MIC), and the peri-implant bone density (BD). RESULTS Despite being present on all tested implant surfaces, MNGCs were not associated with an inflammatory cell infiltrate or with fibrous encapsulation. MNGCs were less numerous on the Ti implants (range: 3.9-5.2%) compared with the ceramic implants (range: 17.6-30.3%, p

AIM: Chemical decontamination increases the availability of bone grafts; however, it is unclear whether antiseptic processing changes the biological activity of bone. MATERIALS AND METHODS: Bone chips were incubated with 4 different antiseptic solutions including (1) povidone-iodine (0.5%), (2) chlorhexidine diguluconate (0.2%), (3) hydrogen peroxide (1%) and (4) sodium hypochlorite (0.25%). After 10 minutes of incubation, changes in the capacity of the bone-conditioned medium to modulate gene expression of gingival fibroblasts was investigated. RESULTS: Conditioned medium obtained from freshly prepared bone chips increased the expression of TGF-β target genes interleukin 11 (IL11), proteoglycan4 (PRG4), NADPH oxidase 4 (NOX4), and decreased the expression of adrenomedullin (ADM), and pentraxin 3 (PTX3) in gingival fibroblasts. Incubation of bone chips with 0.2% chlorhexidine, followed by vigorously washing resulted in a bone-conditioned medium with even higher expression of IL11, PRG4, and NOX4. These findings were also found with a decrease in cell viability and an activation of apoptosis signaling. Chlorhexidine alone, at low concentrations, increased IL11, PRG4 and NOX4 expression, independent of the TGF-β receptor I kinase activity. In contrast, 0.25% sodium hypochlorite almost entirely abolished the activity of bone-conditioned medium, while the other two antiseptic solutions, 1% hydrogen peroxide and 0.5% povidone-iodine, had relatively no impact, respectively. CONCLUSION: These in vitro findings demonstrate that incubation of bone chips with chlorhexidine differentially affects the activity of the respective bone-conditioned medium compared to the other antiseptic solutions. The data further suggest that the main effects are caused by chlorhexidine remaining in the bone-conditioned medium after repeated washing of the bone chips. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved. KEYWORDS: Autografts; TGF-β; antiseptic solution; bone; bone conditioned medium; bone supernatant; chlorhexidine; hydrogen peroxide; povidone-iodine; sodium hypochlorite

The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not.

OBJECTIVES Since the original description of osteoinduction in the early 20th century, the study and development of innovative biomaterials has emerged. Recently, novel synthetic bone grafts have been reported with potential to form ectopic bone in vivo. However, their full characterization in comparison with other leading bone grafts has not been investigated. The aim of this study was to determine the osteoinductive potential of bone grafts by comparing autogenous bone grafts, demineralized freeze-dried bone allografts (DFDBA), a commonly utilized natural bone mineral (NBM) from bovine origin (Bio-Oss), and a newly developed biphasic calcium phosphate (BCP). MATERIALS AND METHODS Grafts were compared in vitro for their ability to stimulate bone marrow stromal cell (BMSC) migration, proliferation, and differentiation as assessed by quantitative real-time PCR for genes coding for bone markers including Runx2, collagen I, and osteocalcin. Furthermore, bone grafts were implanted in the calf muscle of 12 beagle dogs to determine their potential to form ectopic bone in vivo. RESULTS The in vitro results demonstrate that both autografts and DFDBA show potential for cell recruitment, whereas only autografts and BCP demonstrated the ability to differentiate BMSCs toward the osteoblast lineage. The in vivo ectopic bone model demonstrated that while NBM particles were not osteoinductive and autogenous bone grafts were resorbed quickly in vivo, ectopic bone formation was reported in DFDBA and in synthetic BCP grafts. CONCLUSION The modifications in nanotopography and chemical composition of the newly developed BCP bone grafts significantly promoted ectopic bone formation confirming their osteoinductive potential. In conclusion, the results from this study provide evidence that synthetic bone grafts not only serve as a three-dimensional scaffold but are also able to promote osteoinduction.

PURPOSE Autologous bone is used for augmentation in the course of oral implant placement. Bone grafts release paracrine signals that can modulate mesenchymal cell differentiation in vitro. The detailed genetic response of the bone-derived fibroblasts to these paracrine signals has remained elusive. Paracrine signals accumulate in bone-conditioned medium (BCM) prepared from porcine cortical bone chips. MATERIALS AND METHODS In this study, bone-derived fibroblasts were exposed to BCM followed by a whole genome expression profiling and downstream quantitative reverse transciptase polymerase chain reaction of the most strongly regulated genes. RESULTS The data show that ADM, IL11, IL33, NOX4, PRG4, and PTX3 were differentially expressed in response to BCM in bone-derived fibroblasts. The transforming growth factor beta (TGF-β) receptor 1 antagonist SB431542 blocked the effect of BCM on the expression of the gene panel, except for IL33. CONCLUSION These in vitro results extend existing evidence that cortical bone chips release paracrine signals that provoke a robust genetic response in mesenchymal cells that is not exclusively mediated via the TGF-β receptor. The present data provide further insights into the process of graft consolidation.

PURPOSE Whole saliva comprises components of the salivary pellicle that spontaneously forms on surfaces of implants and teeth. However, there are no studies that functionally link the salivary pellicle with a possible change in gene expression. MATERIALS AND METHODS This study examined the genetic response of oral fibroblasts exposed to the salivary pellicle and whole saliva. Oral fibroblasts were seeded onto a salivary pellicle and the respective untreated surface. Oral fibroblasts were also exposed to freshly harvested sterile-filtered whole saliva. A genome-wide microarray of oral fibroblasts was performed, followed by gene ontology screening with DAVID functional annotation clustering, KEGG pathway analysis, and the STRING functional protein association network. RESULTS Exposure of oral fibroblasts to saliva caused 61 genes to be differentially expressed (P < .05). Gene ontology screening assigned the respective genes into 262 biologic processes, 3 cellular components, 13 molecular functions, and 7 pathways. Most remarkable was the enrichment in the inflammatory response. None of the genes regulated by whole saliva was significantly changed when cells were placed onto a salivary pellicle. CONCLUSION The salivary pellicle per se does not provoke a significant inflammatory response of oral fibroblasts in vitro, whereas sterile-filtered whole saliva does produce a strong inflammatory response.

OBJECTIVES Osteoclasts rapidly form on the surface of bone chips at augmentation sites. The underlying molecular mechanism, however, is unclear. Soluble factors released from bone chips in vitro have a robust impact on mesenchymal cell differentiation. Whether these soluble factors change the differentiation of hematopoietic cells into osteoclasts remains unknown. METHODS Osteoclastogenesis, the formation of tartrate-resistant acid phosphatase-positive multinucleated cells, was studied with murine bone marrow cultures exposed to RANKL and M-CSF, and conditioned medium from fresh (BCM) and demineralized bone matrix (DCM). Histochemical staining, gene and protein expression, as well as viability assays were performed. RESULTS This study shows that BCM had no impact on osteoclastogenesis. However, when BCM was heated to 85°C (BCMh), the number of tartrate-resistant acid phosphatase-positive multinucleated cells that developed in the presence of RANKL and M-CSF approximately doubled. In line with the histochemical observations, there was a trend that BCMh increased expression of osteoclast marker genes, in particular the transcription factor c-fos. The expression of c-fos was significantly reduced by the TGF-β receptor I antagonist SB431542. DCM significantly stimulated osteoclastogenesis, independent of thermal processing. CONCLUSIONS These data demonstrate that activated BCM by heat and DBM are able to stimulate osteoclastogenesis in vitro. These in vitro results support the notion that the resorption of autografts may be supported by as yet less defined paracrine mechanisms.