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OBJECTIVES Collagen membranes are not limited to be occlusive barriers as they actively support bone regeneration. However, the impact of bone-derived growth factors on their osteoconductive competence has not been examined. METHODS Twenty adult Sprague Dawley rats were included in the study. Calvaria defects with a diameter of five millimeter were created. The defect was covered with one layer of a collagen membrane previously soaked in conditioned medium of porcine bone chips or in culture medium alone. After 4 weeks, microcomputed tomography was performed. Undecalcified thin-ground sections were subjected to light and scanning electron microscopy. Primary outcome parameter was the bone volume in the defect. Unit of analysis was the bone-conditioned medium (BCM). RESULTS In the central defect area of the control and the BCM group, median new bone connected to the host bone was 0.54 and 0.32 mm³, respectively (p = .10). In the ectocranial defect area, the control group showed significantly more bone than the BCM group (0.90 and 0.26 mm³; p = .02). Based on an exploratory interpretation, the control group had smaller bony islands than the BCM group. Scanning electron microscopy and histology indicate the formation of bone but also the collagen membrane to be mineralized in the defect site. CONCLUSIONS These results demonstrate that the commercial collagen membrane holds an osteoconductive competence in a rat calvaria defect model. Soaking collagen membranes with BCM shifts bone formation toward the formation of bony islands rather than new bone connected to the host bone.

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.

Made available in DSpace on 2018-12-11T17:31:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01 Objectives: Alendronate and raloxifene, a bisphosphonate and a selective estrogen modulator, respectively, are established osteoporosis therapies. Current evidence suggests that simultaneous application of osteoporosis therapies modulates osseointegration. However, alendronate shows inconsistent findings and raloxifene has not been studied comprehensively. This study aimed to evaluate the bone dynamics and molecular and microstructural features at the peri-implant bone interface in osteoporotic rats. Materials and methods: Thirty female rats underwent ovariectomy and were fed a diet low in calcium and phosphate and treated with alendronate or raloxifene for 30 days or underwent fictional ovariectomy surgery (SHAM) prior to implant insertion in the tibia; osteoporosis therapies continued thereafter. After 42 days, peri-implant bone was evaluated by histometric and micro-CT analysis. Fluorochrome incorporation and gene expression was determined to evaluate bone turnover. Results: We report here that alendronate had no impact on bone-to-implant contacts and the mineral apposition rate. The RANKL/OPG ratio and local bone volume, however, were increased compared to the untreated osteoporotic rats. Even though signaling to bone resorption activity through RANKL production was observed in the alendronate group, the blockade of bone resorption activity that occurs in decorrence to alendronate activity took place and resulted in an increase in bone volume. Raloxifene significantly increased osseointegration in osteoporotic rats, as indicated by bone-to-implant contacts, mineral apposition, and local bone volume. Raloxifene, however, had no considerable impact on the RANKL/OPG ratio compared to untreated osteoporotic rats. As expected, the SH group showed higher bone-to-implant contacts and mineral apposition rates than the untreated osteoporotic rats. Conclusions: These findings suggest that raloxifene but not alendronate can compensate for the impaired osseointegration in osteoporotic rats. Clinical relevance: Regarding the superiority of raloxifene observed in the improvement of bone dynamics response, this statement suggests that raloxifene could be a good option for osteoporosis patients in oral rehabilitation procedures. Department of Surgery and Integrated Clinic Division of Oral and Maxillofacial Surgery São Paulo State University (UNESP) School of Dentistry Department of Basic Sciences São Paulo State University (UNESP) School of Dentistry Department of Morphology Piracicaba Dental School University of Campinas (UNICAMP) Department of Surgery and Integrated Clinic Division of Integrated Clinic São Paulo State University UNESP) School of Dentistry Department of Oral Biology Medical University of Vienna Department of Surgery and Integrated Clinic Division of Oral and Maxillofacial Surgery São Paulo State University (UNESP) School of Dentistry Department of Basic Sciences São Paulo State University (UNESP) School of Dentistry Department of Surgery and Integrated Clinic Division of Integrated Clinic São Paulo State University UNESP) School of Dentistry

Tooth development commences at the end of the fifth week of human gestation. It is commonly divided into the following stages: the initiation stage, the bud stage, the cap stage, the bell stage, and finally maturation. The aim of this chapter is to describe the process of early tooth development, through maturation and culminating in coronal formation. The morphology of pits and fissures is characterized showing serial ground sections and micro-CTs.

Objective Milk is a rich source of transforming growth factor (TGF)-β which supports intestinal mucosal homeostasis of infants. Milk may also have beneficial effects on the integrity of the oral cavity, its being part of the gastrointestinal tract. However, it is unclear if milk and fermented milk products provoke a TGF-β response in oral cells. Material and Methods Human gingival fibroblasts were exposed to pasteurized cow milk, yoghurt, sour milk, buttermilk and whey, followed by a reverse transcriptase polymerase chain reaction of the TGF-β target genes interleukin11 (IL11), proteoglycan4 (PRG4), and NADPH oxidase 4 (NOX4). Immunoassays were performed for IL11 and TGF-β in cell culture supernatant and milk products, respectively. Signaling was investigated with the TGF-β receptor type I kinase inhibitor SB431542. Results We report here that pasteurized cow milk and the aqueous fractions of yoghurt, sour milk, buttermilk and whey enhanced the expression of IL11, NOX4 and PRG4 in gingival fibroblasts. Moreover, IL11 protein levels in the respective supernatant were significantly increased. Cow milk, yoghurt, sour milk and buttermilk contain approximately 1–2 ng TGF-β1, whereas active TGF-β1 is hardly detectable in whey. SB431542 reduced the response of gingival fibroblasts to pasteurized cow milk and fermented milk products based on IL11 release into the supernatant. Conclusions These results demonstrate that gingival fibroblasts respond to pasteurized cow milk and to fermented milk products with an increased expression of TGF-β target genes.

Aim Tumour necrosis factor (TNF)-α is a pathological factor causing the characteristic symptoms of periodontal disease and rheumatoid arthritis. In this study, we describe the phenotypes of human TNF-α transgenic mice (hTNFtg) with respect to their periodontium and the temporomandibular joint (TMJ). Material and methods Periodontal structures, TMJ and skull shape of hTNFtg mice and wild-type (WT) littermates were assessed by microcomputed tomography, automated segmentation, geometric morphometrics and histologic ground sections. Results We show that hTNFtg mice have an eroded lamina dura and reduced periodontal ligament space compared to (WT) littermates. Transgenic mice further exhibit severe destruction of the TMJ. Geometric morphometrics revealed that hTNFtg mice have a more laterally positioned TMJ with a concomitantly enlarged zygomatic process. Mandibular and maxillary teeth occluded properly. Conclusions Our findings suggest that chronic inflammation in hTNFtg mice causes destructive changes of the periodontium and the TMJ.

While oral rinses used for cosmetic purposes only do not necessarily have to be antiseptic, antimicrobial activity is required for medical indications, including oral and periodontal surgery. So the question arises—is the antimicrobial activity of oral rinses associated with any destructive changes in cell viability in vitro? To answer this question, we examined twelve oral rinses with respect to their antimicrobial and cytotoxic activity. Antimicrobial activity was screened against five bacterial strains using disc diffusion. Cytotoxicity was determined by mitochondrial reductase activity with primary gingival fibroblasts, L929 cells, and HSC-2 epithelial cells. Phase contrast microscopy and trypan blue staining were then performed to reveal cell morphology. Cells remained vital after exposure to oral rinses that were only used for cosmetic purposes. Moderate cytotoxic effects were observed for oral rinses containing 0.05% chlorhexidine, ethanol, or pegylated hydrogenated castor oil and sodium dodecyl sulfate. Other oral rinses containing 0.2% chlorhexidine and cocamidopropyl betaine exhibited strong cytotoxic and antimicrobial activity. Strong cytotoxic but moderate antimicrobial activity was observed in oral rinses containing cetylpyridinium chloride. The in vitro data show that oral rinses are heterogeneous with respect to their cytotoxic and antimicrobial effects. Based on their respective properties, oral rinses can be selected either to reduce the microbial load or for cosmetic purposes.

BACKGROUND AND OBJECTIVE Inhibition of prolyl hydroxylases stimulates bone regeneration. Consequently, bone substitute materials were developed that release prolyl hydroxylase inhibitors. However, the impact of prolyl hydroxylase inhibitors released from these carriers on osteoclastogenesis is not clear. We therefore assessed the effect of bone substitute materials that release prolyl hydroxylase inhibitors on osteoclastogenesis. MATERIAL AND METHODS Dimethyloxalylglycine, desferrioxamine, and l-mimosine were lyophilized onto bovine bone mineral and hydroxyapatite, and supernatants were generated. Osteoclastogenesis was induced in murine bone marrow cultures in the presence of the supernatants from bone substitute materials. The formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells and TRAP activity were determined. To test for possible effects on osteoclast progenitor cells, we measured the effect of the supernatants on proliferation and viability. In addition, experiments were performed where prolyl hydroxylase inhibitors were directly added to the bone marrow cultures. RESULTS We found that prolyl hydroxylase inhibitors released within the first hours from bone substitute materials reduce the number and activity of TRAP-positive multinucleated cells. In line with this, addition of prolyl hydroxylase inhibitors directly to the bone marrow cultures dose-dependently reduced the number of TRAP-positive multinucleated cells and the overall resorption activity. Moreover, the released prolyl hydroxylase inhibitors decreased proliferation but not viability of osteoclast progenitor cells. CONCLUSION Our results show that prolyl hydroxylase inhibitors released from bone substitute materials decrease osteoclastogenesis in murine bone marrow cultures.

OBJECTIVE Prolyl hydroxylases (PHD) are oxygen sensors and therefore pharmacological targets to stimulate periodontal regeneration. Here we evaluate the release profile of the PHD inhibitors dimethyloxaloylglycine and l-mimosine from bone substitutes. MATERIALS Dimethyloxaloylglycine and l-mimosine were lyophilised onto bone substitutes including bovine bone mineral, beta-tricalcium phosphate, and hydroxyapatite. Release kinetic was evaluated by bioassays with gingival and periodontal ligament fibroblasts. We determined the capacity of PHD inhibitors to provoke VEGF expression and to repress metabolic activity and proliferation as assessed by immunoassay, MTT conversion and (3)[H]thymidine incorporation, respectively. RESULTS We found that the PHD inhibitors are released from bovine bone mineral as indicated by the increase of VEGF production in gingival and periodontal ligament fibroblasts. Supernatants obtained after 1h also decreased metabolic activity and proliferation of the fibroblasts. A fibrin matrix prolonged the release of PHD inhibitors up to 192h. A similar cellular response was found when supernatants from PHD inhibitors loaded beta-tricalcium phosphate and hydroxyapatite embedded in fibrin were assessed. CONCLUSIONS In conclusion bone substitutes can serve as carriers for PHD inhibitors that maintain their capacity to provoke a pro-angiogenic response in vitro. These findings provide the basis for preclinical studies to evaluate if this release kinetic can stimulate periodontal regeneration.

PURPOSE Autografts are considered to support bone regeneration. Paracrine factors released from cortical bone might contribute to the overall process of graft consolidation. The aim of this study was to characterize the paracrine factors by means of proteomic analysis. MATERIALS AND METHODS Bone-conditioned medium (BCM) was prepared from fresh bone chips of porcine mandibles and subjected to proteomic analysis. Proteins were categorized and clustered using the bioinformatic tools UNIPROT and PANTHER, respectively. RESULTS Proteomic analysis showed that BCM contains more than 150 proteins, of which 43 were categorized into "secreted" and "extracellular matrix." Growth factors that are not only detectable in BCM, but potentially also target cellular processes involved in bone regeneration, eg, pleiotrophin, galectin-1, transforming growth factor beta (TGF-β)-induced gene (TGFBI), lactotransferrin, insulin-like growth factor (IGF)-binding protein 5, latency-associated peptide forming a complex with TGF-β1, and TGF-β2, were discovered. CONCLUSION The present results demonstrate that cortical bone chips release a large spectrum of proteins with the possibility of modulating cellular aspects of bone regeneration. The data provide the basis for future studies to understand how these paracrine factors may contribute to the complex process of graft consolidation.

To assess the pro-angiogenic and pro-inflammatory capacity of the dentine-pulp complex in response to the prolyl hydroxylase inhibitor L-mimosine in a tooth slice organ culture model.Human teeth were sectioned transversely into 600-μm-thick slices and cultured in medium supplemented with serum and antibiotics. Then, pulps were stimulated for 48 h with L-mimosine. Pulps were subjected to viability measurements based on formazan formation in MTT assays. In addition, histological evaluation of pulps was performed based on haematoxylin and eosin staining. Culture supernatants were subjected to immunoassays for vascular endothelial growth factor (VEGF) to determine the pro-angiogenic capacity and to immunoassays for interleukin (IL)-6 and IL-8 to assess the pro-inflammatory response. Interleukin-1 served as pro-inflammatory control. Echinomycin was used to inhibit hypoxia-inducible factor-1 (HIF-1) alpha activity. Data were analysed using Student's t-test and Mann-Whitney U test.Pulps within tooth slices remained vital upon L-mimosine stimulation as indicated by formazan formation and histological evaluation. L-mimosine increased VEGF production when normalized to formazan formation in the pulp tissue of the tooth slices (P 0.05). This effect on VEGF was reduced by echinomycin (P 0.01). Changes in normalized IL-6 and IL-8 levels upon treatment with L-mimosine did not reach the level of significance (P 0.05), whilst treatment with IL-1, which served as positive control, increased IL-6 (P 0.05) and IL-8 levels (P 0.05).The prolyl hydroxylase inhibitor L-mimosine increased VEGF production via HIF-1 alpha in the tooth slice organ culture model whilst inducing no prominent increase in IL-6 and IL-8. Pre-clinical studies will reveal if these in vitro effects translate into dental pulp regeneration.

BACKGROUND AND OBJECTIVE Connective tissue grafts are frequently applied, together with Emdogain(®) , for root coverage. However, it is unknown whether fibroblasts from the gingiva and from the palate respond similarly to Emdogain. The aim of this study was therefore to evaluate the effect of Emdogain(®) on fibroblasts from palatal and gingival connective tissue using a genome-wide microarray approach. MATERIAL AND METHODS Human palatal and gingival fibroblasts were exposed to Emdogain(®) and RNA was subjected to microarray analysis followed by gene ontology screening with Database for Annotation, Visualization and Integrated Discovery functional annotation clustering, Kyoto Encyclopedia of Genes and Genomes pathway analysis and the Search Tool for the Retrieval of Interacting Genes/Proteins functional protein association network. Microarray results were confirmed by quantitative RT-PCR analysis. RESULTS The transcription levels of 106 genes were up-/down-regulated by at least five-fold in both gingival and palatal fibroblasts upon exposure to Emdogain(®) . Gene ontology screening assigned the respective genes into 118 biological processes, six cellular components, eight molecular functions and five pathways. Among the striking patterns observed were the changing expression of ligands targeting the transforming growth factor-beta and gp130 receptor family as well as the transition of mesenchymal epithelial cells. Moreover, Emdogain(®) caused changes in expression of receptors for chemokines, lipids and hormones, and for transcription factors such as SMAD3, peroxisome proliferator-activated receptor gamma and those of the ETS family. CONCLUSION The present data suggest that Emdogain(®) causes substantial alterations in gene expression, with similar patterns observed in palatal and gingival fibroblasts.

OBJECTIVE Short implants are increasingly used, but there is doubt about their performance being similar to that of regular implants. The aim of this study was to compare the mechanical stability of short implants vs. regular implants placed in the edentulous posterior mandible. MATERIAL AND METHODS Twenty-three patients received a total of 48 short implants (5 × 5.5 mm and 5 × 7 mm) and 42 regular implants (4 × 10 mm and 4 × 11.5 mm) in the posterior mandible. Patients who received short implants had

Sclerostin is a Wnt signalling antagonist that controls bone metabolism. Sclerostin is expressed by osteocytes and cementocytes; however, its role in the formation of dental structures remains unclear. Here, we analysed the mandibles of sclerostin knockout mice to determine the influence of sclerostin on dental structures and dimensions using histomorphometry and micro-computed tomography (μCT) imaging. μCT and histomorphometric analyses were performed on the first lower molar and its surrounding structures in mice lacking a functional sclerostin gene and in wild-type controls. μCT on six animals in each group revealed that the dimension of the basal bone as well as the coronal and apical part of alveolar part increased in the sclerostin knockout mice. No significant differences were observed for the tooth and pulp chamber volume. Descriptive histomorphometric analyses of four wild-type and three sclerostin knockout mice demonstrated an increased width of the cementum and a concomitant moderate decrease in the periodontal space width. Taken together, these results suggest that the lack of sclerostin mainly alters the bone and cementum phenotypes rather than producing abnormalities in tooth structures such as dentin.

The use of prolyl hydroxylase inhibitors such as l-mimosine (L-MIM) and dimethyloxaloylglycine (DMOG) to improve angiogenesis is a new approach for periodontal regeneration. In addition to exhibiting pro-angiogenic effects, prolyl hydroxylase inhibitors can modulate the plasminogen activator system in cells from non-oral tissues. This study assesses the effect of prolyl hydroxylase inhibitors on plasminogen activation by fibroblasts from the periodontium.Gingival and periodontal ligament fibroblasts were incubated with L-MIM and DMOG. To investigate whether prolyl hydroxylase inhibitors modulate the net plasminogen activation, kinetic assays were performed with and without interleukin (IL)-1. Moreover, plasminogen activators and the respective inhibitors were analyzed by casein zymography, immune assays, and quantitative polymerase chain reaction.The kinetic assay showed that L-MIM and DMOG reduced plasminogen activation under basal and IL-1-stimulated conditions. Casein zymography revealed that the effect of L-MIM involves a decrease in urokinase-type plasminogen activator activity. In agreement with these findings, reduced levels of urokinase-type plasminogen activator and elevated levels of plasminogen activator inhibitor 1 were observed.L-MIM and DMOG can reduce plasminogen activation by fibroblasts from the gingiva and the periodontal ligament under basal conditions and in the presence of an inflammatory cytokine.

Aim To provide an overview on the biology and soft tissue wound healing around teeth and dental implants. Material and Methods This narrative review focuses on cell biology and histology of soft tissue wounds around natural teeth and dental implants. Results and conclusions The available data indicate that: Oral wounds follow a similar pattern. The tissue specificities of the gingival, alveolar and palatal mucosa appear to be innately and not necessarily functionally determined. The granulation tissue originating from the periodontal ligament or from connective tissue originally covered by keratinized epithelium has the potential to induce keratinization. However, it also appears that deep palatal connective tissue may not have the same potential to induce keratinization as the palatal connective tissue originating from an immediately subepithelial area. Epithelial healing following non-surgical and surgical periodontal therapy appears to be completed after a period of 7–14 days. Structural integrity of a maturing wound between a denuded root surface and a soft tissue flap is achieved at approximately 14-days post-surgery. The formation of the biological width and maturation of the barrier function around transmucosal implants requires 6–8 weeks of healing. The established peri-implant soft connective tissue resembles a scar tissue in composition, fibre orientation, and vasculature. The peri-implant junctional epithelium may reach a greater final length under certain conditions such as implants placed into fresh extraction sockets versus conventional implant procedures in healed sites.

BACKGROUND: Harvesting techniques can affect cellular parameters of autogenous bone grafts in vitro. Whether these differences translate to in vivo bone formation, however, remains unknown. OBJECTIVE: The purpose of this study was to assess the impact of different harvesting techniques on bone formation and graft resorption in vivo. MATERIAL AND METHODS: Four harvesting techniques were used: (i) corticocancellous blocks particulated by a bone mill; (ii) bone scraper; (iii) piezosurgery; and (iv) bone slurry collected from a filter device upon drilling. The grafts were placed into bone defects in the mandibles of 12 minipigs. The animals were sacrificed after 1, 2, 4 and 8 weeks of healing. Histology and histomorphometrical analyses were performed to assess bone formation and graft resorption. An explorative statistical analysis was performed. RESULTS: The amount of new bone increased, while the amount of residual bone decreased over time with all harvesting techniques. At all given time points, no significant advantage of any harvesting technique on bone formation was observed. The harvesting technique, however, affected bone formation and the amount of residual graft within the overall healing period. Friedman test revealed an impact of the harvesting technique on residual bone graft after 2 and 4 weeks. At the later time point, post hoc testing showed more newly formed bone in association with bone graft processed by bone mill than harvested by bone scraper and piezosurgery. CONCLUSIONS: Transplantation of autogenous bone particles harvested with four techniques in the present model resulted in moderate differences in terms of bone formation and graft resorption.

Cornifelin (CNFN) has been identified as a protein component of epidermal corneocytes. Here, we investigated the tissue distribution of CNFN and potential consequences of CNFN deficiency on epithelial function in in vitro models of human skin and oral mucosa. Our detailed bioinformatics and immunostaining analysis revealed that CNFN is not only expressed in human epidermis but also in noncornifying oral mucosa. In normal epidermis, CNFN was confined to the upper granular layer and the stratum corneum. By contrast, in both partly cornifying and noncornifying oral mucosa, CNFN was expressed in a cell membrane-associated pattern over several suprabasal layers. Small interfering RNA-mediated knockdown of CNFN in epidermal keratinocytes (KCs) was associated with only subtle alterations of the overall epidermal architecture in skin models in vitro but led to altered morphology of corneodesmosomes, as detected by electron microscopy. Using dispase treatment followed by mechanical stress, epithelial sheets of CNFN-deficient epidermal KCs were easily disrupted, whereas their CNFN-competent counterparts remained intact. In contrast to the epidermal KCs, CNFN knockdown in oral KCs had a more severe effect and caused pronounced acantholysis in organotypic models of oral mucosa. Together, these findings indicate that CNFN is a structural component of the cell adhesion system of differentiated KCs in both epidermis and oral mucosa.

BACKGROUND AND PURPOSE Autografts are used for bone reconstruction in regenerative medicine including oral and maxillofacial surgery. Bone grafts release paracrine signals that can reach mesenchymal cells at defect sites. The impact of the paracrine signals on osteogenic, adipogenic, and chondrogenic differentiation of mesenchymal cells has remained unclear. MATERIAL AND METHODS Osteogenesis, adipogenesis, and chondrogenesis were studied with murine ST2 osteoblast progenitors, 3T3-L1 preadipocytes, and ATDC5 prechondrogenic cells, respectively. Primary periodontal fibroblasts from the gingiva, from the periodontal ligament, and from bone were also included in the analysis. Cells were exposed to bone-conditioned medium (BCM) that was prepared from porcine cortical bone chips. RESULTS BCM inhibited osteogenic and adipogenic differentiation of ST2 and 3T3-L1 cells, respectively, as shown by histological staining and gene expression. No substantial changes in the expression of chondrogenic genes were observed in ATDC5 cells. Primary periodontal fibroblasts also showed a robust decrease in alkaline phosphatase and peroxisome proliferator-activated receptor gamma (PPARγ) expression when exposed to BCM. BCM also increased collagen type 10 expression. Pharmacologic blocking of transforming growth factor (TGF)-β receptor type I kinase with SB431542 and the smad-3 inhibitor SIS3 at least partially reversed the effect of BCM on PPARγ and collagen type 10 expression. In support of BCM having TGF-β activity, the respective target genes were increasingly expressed in periodontal fibroblasts. CONCLUSIONS The present work is a pioneer study on the paracrine activity of bone grafts. The findings suggest that cortical bone chips release soluble signals that can modulate differentiation of mesenchymal cells in vitro at least partially involving TGF-β signaling.

Platelets modulate formation of osteoclasts and osteoblasts, but research with different preparations of platelets remains inconclusive. Here, we assessed whether serum components modulate the effect of platelet preparations. In murine bone marrow cultures, osteoclastogenesis was investigated in the presence of platelet-released supernatant (PRS), serum containing PRS (SC-PRS), and serum. Osteoclastogenesis was quantified by the numbers of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells, TRAP activity and resorption assays. Also human osteoclastogenesis assays were performed. Viability and proliferation were tested by MTT and (3) [H]thymidine incorporation assays, respectively. Osteoblastogenesis was assessed by histochemical staining for alkaline phosphatase-of murine bone marrow cultures and human MG63 cells. We found PRS to increase the number of TRAP(+) multinucleated cells in the early phase and TRAP activity in the later phase of osteoclastogenesis. SC-PRS and serum decreased the number and activity of TRAP(+) multinucleated cells. Both serum containing preparations reduced viability and proliferation of hematopoietic progenitors. PRS decreased the numbers of alkaline phosphatase-positive colonies while SC-PRS and serum increased osteoblastmarkers in MG63. Proliferation of MG63 was stimulated by all preparations. These results show that activated platelets support osteoclastogenesis, while platelet preparations that contain serum components decrease osteoclastogenesis and increase osteoblastogenesis in vitro, suggesting that serum components modulate the effects of platelets on osteoclastogenesis and osteoblastogenesis.

BACKGROUND: Zirconia (ZrO2 ) has received interest as a dental material; however, little information is available on the impact of surface modifications on the osseointegration of zirconia implants. PURPOSE: The aim of the present study was to determine the effect of acid or alkaline etching of sandblasted ZrO2 implants on bone apposition in vivo. METHODS: Cylindrical ZrO2 implants with two circumferential grooves were placed in the maxilla of 12 miniature pigs. Biopsies were harvested after 1, 2, 4, and 8 weeks of healing. Undecalcified toluidine blue-stained ground sections were produced. The bone-to-implant contact, the bone area, and the presence of multinucleated giant cells were determined by histomorphometry. An uncorrected explorative statistical analysis was performed. RESULTS: Acid etching but not alkaline etching of sandblasted ZrO2 implants caused more bone-to-implant contact than sandblasted ZrO2 implants. The bone area was unaffected by the surface modifications. Acid and alkaline etching both increased the formation of multinucleated giant cells at the implant surface. CONCLUSIONS: This study provides a scientific basis to further investigate the impact of acid etching of sandblasted ZrO2 implants on osseointegration and the role of multinucleated giant cells in this process.

Objectives Saliva can suppress osteoclastogenesis, but the underlying mechanism has not been discovered yet. Considering that endotoxins suppress osteoclastogenesis in bone marrow cultures and that saliva contains endotoxins, it was reasonable to hypothesize that the impact of saliva on osteoclastogenesis requires toll-like receptor 4 signaling. Material and methods To test this hypothesis, we blocked toll-like receptor 4 signaling with TAK-242 in the presence of saliva in murine bone marrow cultures. Osteoclastogenesis was evaluated based on gene expression analysis and histochemical staining for tartrate-resistant acid phosphatase. Resorption was performed on dentine. Results We report that TAK-242 reversed the inhibitory effect of fresh sterile saliva on the formation of multinucleated cells that stained positive for tartrate-resistant acid phosphatase. In line with this finding, TAK-242 increased the expression of the osteoclast functional genes cathepsin K, calcitonin receptor, and tartrate-resistant acid phosphatase in the presence of saliva. TAK-242 also supported the expression of NFATc1, the master regulator of osteoclastogenesis, as well as DC-STAMP and Atp6v0d2, both being cell fusion genes. In support of the hypothesis, depletion of saliva from endotoxin partially reversed the inhibitory effect on osteoclastogenesis. Moreover, salivary pellicle on plastic and titanium did not affect osteoclastogenesis. Conclusion Inhibition of toll-like receptor 4 signaling revealed that saliva can contribute to innate immunity by preventing hematopoietic progenitors to become osteoclasts. Clinical relevance Saliva can activate pattern recognition receptor signaling through endotoxins and other stress factors, indicating the demand for macrophages rather than for osteoclasts. Electronic supplementary material The online version of this article (doi:10.1007/s00784-016-2041-7) contains supplementary material, which is available to authorized users.

Preclinical and clinical studies suggest that a combination of enamel matrix derivative (EMD) with demineralized freeze-dried bone allograft (DFDBA) may improve periodontal wound healing and regeneration. To date, no single study has characterized the effects of this combination on in vitro cell behavior. The aim of this study is to test the ability of EMD to adsorb to the surface of DFDBA particles and determine the effect of EMD coating on downstream cellular pathways such as adhesion, proliferation, and differentiation of primary human osteoblasts and periodontal ligament (PDL) cells.DFDBA particles were precoated with EMD or human blood and analyzed for protein adsorption patterns via scanning electron microscopy. Cell attachment and proliferation were quantified using a commercial assay. Cell differentiation was analyzed using real-time polymerase chain reaction for genes encoding Runx2, alkaline phosphatase, osteocalcin, and collagen 1α1, and mineralization was assessed using alizarinred staining.Analysis of cell attachment revealed no significant differences among control, blood-coated, and EMD-coated DFDBA particles. EMD significantly increased cell proliferation at 3 and 5 days after seeding for both osteoblasts and PDL cells compared to control and blood-coated samples. Moreover, there were significantly higher messenger ribonucleic acid levels of osteogenic differentiation markers, including collagen 1α1, alkaline phosphatase, and osteocalcin, in osteoblasts and PDL cells cultured on EMD-coated DFDBA particles at 3, 7, and 14 days.The results suggest that the addition of EMD to DFDBA particles may influence periodontal regeneration by stimulating PDL cell and osteoblast proliferation and differentiation.

BACKGROUND AND OBJECTIVE In regenerative dentistry, platelet preparations are applied to stimulate bone healing and periodontal regeneration. Here, we pursue a strategy where bone substitutes are used as carriers for platelet-released supernatants. The mitogenic capacity and release kinetics of loaded bone substitutes were assessed. MATERIAL AND METHODS Platelet-released supernatants of washed platelets (washed PRS) and platelet-released supernatants of unwashed platelets (unwashed PRS) were lyophilized onto the bone substitutes deproteinized bovine bone mineral, hydroxyapatite and β-tricalcium phosphate. Scanning electron microscopy images were taken. Supernatants of bone substitutes were collected at hours 1, 3, 6, 24, and 48 and medium was replaced. We evaluated the protein content with the bicinchoninic acid assay and the effect on proliferation using bioassays with human periodontal fibroblasts. Release of growth factors from the loaded bone substitutes was measured based on the platelet-derived growth factor isoform (PDGF-BB) and thrombin immunoassays. Furthermore, we assessed DNA and RNA content of washed PRS and unwashed PRS. RESULTS Unwashed PRS showed higher total protein concentrations than washed PRS, while the concentration of PDGF-BB, thrombin, DNA, RNA and their mitogenic effect was not significantly different. The bone substitute materials adsorbed protein over time but no significant changes in overall appearance was found. Supernatants collected from unwashed PRS-loaded bone substitute after 1 h induced a potent mitogenic response in periodontal fibroblasts. This pro-mitogenic capacity of the supernatants decreased over the observation period. Supernatants of washed PRS-loaded bone substitutes did not induce a substantial mitogenic effect. Levels of PDGF-BB, thrombin and protein were higher in supernatants of unwashed PRS-loaded bone substitutes than of washed PRS-loaded bone substitutes. CONCLUSION Bone substitutes loaded with unwashed PRS, but not bone substitutes loaded with washed PRS show continuously declining release kinetics. These data suggest that plasma components in platelet preparations can modify the release kinetics profile.

OBJECTIVES Autogenous bone grafting has remained the gold standard for bone augmentation procedures with ability to release growth factors to the surrounding microenvironment. Recent investigations have characterized these specific growth factors released by autogenous bone chips with further isolation into a "bone-conditioned medium" (BCM). The aim of the present investigation was to utilize autologous growth factors from bone chips (BCM) in combination with deproteinized bovine bone mineral (DBBM) and investigate the ability for BCM to enhance osteoblast behavior. MATERIALS AND METHODS Mouse ST2 cells were seeded on (1) DBBM particles alone or (2) DBBM + BCM. Thereafter, samples were compared for cell recruitment, adhesion, proliferation, and real-time PCR for osteoblast differentiation markers including Runx2, collagen 1 alpha 2 (COL1A2), alkaline phosphatase (ALP), and osteocalcin (OCN). Alizarin red staining was used to assess mineralization. RESULTS Coating BCM on DBBM particles improved cell migration of ST2 cells and significantly enhanced a 2-fold increase in cell adhesion. While no significant increase in cell proliferation was observed, BCM significantly increased mRNA levels of COL1A2, ALP, and OCN at 3 days post seeding. Furthermore, a 3-fold increase in alizarin red staining was observed on DBBM particles pre-coated with BCM. CONCLUSION Pre-coating DBBM with BCM enhanced the osteoconductive properties of DBBM by mediating osteoblast recruitment, attachment, and differentiation towards bone-forming osteoblasts. Future animal study is necessary to further characterize the added benefit of BCM as an autogenous growth factor source for combination therapies. CLINICAL RELEVANCE The application of BCM in combination with biomaterials may serve as an autogenous growth factor source for bone regeneration.

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 The use of autogenous bone chips during guided bone regeneration procedures has remained the gold standard for bone grafting due to its excellent combination of osteoconduction, osteoinduction and osteogenesis. Recent protocols established by our group have characterized specific growth factors and cytokines released from autogenous bone that have the potential to be harvested and isolated into bone conditioned media (BCM). Due to the advantageous osteo-promotive properties of BCM, the aims of the present study was to pre-coat collagen barrier membranes with BCM and investigate its effect on osteoblast adhesion, proliferation and differentiation for possible future clinical use. METHODS Scanning electron microscopy (SEM) was first used to qualitative assess BCM protein accumulation on the surface of collagen membranes. Thereafter, undifferentiated mouse ST2 stromal bone marrow cells were seeded onto BioGide porcine derived collagen barrier membranes (control) or barrier membranes pre-coated with BCM (test group). Control and BCM samples were compared for cell adhesion at 8 h, cell proliferation at 1, 3 and 5 days and real-time PCR at 5 days for osteoblast differentiation markers including Runx2, alkaline phosphatase (ALP), osteocalcin (OCN) and bone sialoprotein (BSP). Mineralization was further assessed with alizarin red staining at 14 days post seeding. RESULTS SEM images demonstrated evidence of accumulated proteins found on the surface of collagen membranes following coating with BCM. Analysis of total cell numbers revealed that the additional pre-coating with BCM markedly increased cell attachment over 4 fold when compared to cells seeded on barrier membranes alone. No significant difference could be observed for cell proliferation at all time points. BCM significantly increased mRNA levels of osteoblast differentiation markers including ALP, OCN and BSP at 5 days post seeding. Furthermore, barrier membranes pre-coated with BCM demonstrated a 5-fold increase in alizarin red staining at 14 days. CONCLUSION The results from the present study suggest that the osteoconductive properties of porcine-derived barrier membranes could be further improved by BCM by significantly increasing cell attachment, differentiation and mineralization of osteoblasts in vitro. Future animal testing is required to fully characterize the additional benefits of BCM for guided bone regeneration.

This overview provides insights into the molecular and cellular mechanisms involved in guided bone regeneration, in particular focusing on aspects presented in the 3D movie, Cell-To-Cell Communication in Guided Bone Regeneration. The information presented here is based almost exclusively on genetic mouse models in which single genes can be deleted or overexpressed, even in a specific cell type. This information needs to be extrapolated to humans and related to aspects relevant to graft consolidation under the clinical parameters of guided bone regeneration. The overview follows the ground tenor of the Cell-To-Cell Communication series and focuses on aspects of cell-to-cell communication in bone regeneration and guided bone regeneration. Here, we discuss (1) the role of inflammation during bone regeneration, including (2) the importance of the fibrin matrix, and (3) the pleiotropic functions of macrophages. We highlight (4) the origin of bone-forming osteoblasts and bone-resorbing osteoclasts as well as (5) what causes a progenitor cell to mature into an effector cell. (6) We touch on the complex bone adaptation and maintenance after graft consolidation and (7) how osteocytes control this process. Finally, we speculate on (8) how barrier membranes and the augmentation material can modulate graft consolidation.

Vitamin D plays an essential role in bone metabolism as well as in immunity. Hence, it might affect the development and extent of periodontal disease. The aim of this study was the assessment of 25-hydroxyvitamin D (25(OH)D) status in periodontal disease. Twenty-nine patients with severe periodontal disease and 29 healthy volunteers were recruited in this case-control-study. Serum 25(OH)D levels, Periodontal Probing Depth (PPD), Clinical Attachment Level (CAL), Bleeding on Probing (BOP), Body Mass Index (BMI), and current smoking status and smoking history (packyears) were assessed in all participants. Serum 25(OH)D levels were compared between controls and cases. Multivariable logistic regression was used to determine the odds ratio (OR) and 95 % confidence interval (CI) for periodontal disease in 25(OH)D deficient probands. Patients with periodontal disease presented a significantly higher proportion of deficient 25(OH)D levels (i.e.

Made available in DSpace on 2018-12-11T17:08:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Background: Supplementation of bone substitutes with recombinant platelet-derived growth factor-BB (PDGF-BB) can enhance bone regeneration. The aim of the study was to evaluate the effect of PDGF-BB on bone formation in the presence of ß-tricalcium phosphate and bovine bone mineral matrix in a rat calvaria defect model. Methods: The authors examined 5 mm rat calvarial defects treated with ß-tricalcium phosphate (TCP) or demineralized bovine bone mineral (DBBM) with and without 0.3 mg/ml recombinant PDGF-BB. Calvaria defects were randomly divided into the following treatment groups (n = 5); TCP; TCP plus PDGF-BB; DBBM; DBBM plus PDGF-BB; and untreated empty control. After 45 days, bone formation was evaluated by histomorphometry and fluorescence microscopy. Results: The authors report that the area of newly formed bone was similar between the empty controls and the two bone substitutes, TCP and DBBM. Supplementation of TCP and DBBM with PDGF-BB had no significant impact on bone formation. Fluorochrome staining revealed no visible changes in the pattern of bone formation in defects filled with TCP and DBBM, irrespective of PDGF-BB. Furthermore, supplementation with PDGF-BB did not influence biomaterial degradation. Conclusions: The authors concluded that PDGF-BB had no impact on bone formation and degradation of bone substitutes in the respective rodent models. Thus, possible beneficial effects of PDGF-BB may require other model situations. Department of Surgery and Integrated Clinic Araçatuba Dental School UNESP-Univ Estadual Paulista Karl Donath Laboratory for Hard Tissue and Biomaterial Research Division of Oral Surgery Medical University of Vienna Austrian Cluster for Tissue Regeneration Department of Preventive Restorative and Pediatric Dentistry School of Dental Medicine University of Bern Department of Oral Biology Medical University of Vienna Department of Anatomy Araçatuba Dental School UNESP-Univ Estadual Paulista Department of Surgery and Integrated Clinic Araçatuba Dental School UNESP-Univ Estadual Paulista Department of Anatomy Araçatuba Dental School UNESP-Univ Estadual Paulista

Introduction Prolyl hydroxylase (PHD) inhibitors can induce a proangiogenic response that stimulates regeneration in soft and hard tissues. However, the effect of PHD inhibitors on the dental pulp is unclear. The purpose of this study was to evaluate the effects of PHD inhibitors on the proangiogenic capacity of human dental pulp–derived cells. Methods To test the response of dental pulp–derived cells to PHD inhibitors, the cells were exposed to dimethyloxalylglycine, desferrioxamine, L-mimosine, and cobalt chloride. To assess the response of dental pulp cells to a capping material supplemented with PHD inhibitors, the cells were treated with supernatants from calcium hydroxide. Viability, proliferation, and protein synthesis were assessed by formazan formation, 3 [H]thymidine, and 3 [H]leucine incorporation assays. The effect on the proangiogenic capacity was measured by immunoassays for vascular endothelial growth factor (VEGF). Results We found that all 4 PHD inhibitors can reduce viability, proliferation, and protein synthesis at high concentrations. At nontoxic concentrations and in the presence of supernatants from calcium hydroxide, PHD inhibitors stimulated the production of VEGF in dental pulp–derived cells. When calcium hydroxide was supplemented with the PHD inhibitors, the supernatants from these preparations did not significantly elevate VEGF levels. Conclusions These results show that PHD inhibitors can stimulate VEGF production of dental pulp–derived cells, suggesting a corresponding increase in their proangiogenic capacity. Further studies will be required to understand the impact that this might have on pulp regeneration.

The aim of this study was to compare selected crevicular markers in early wound healing after second stage surgery and immediately restored implant placement. A total of ten patients (mean age 53 years, 6 females and 4 males) to be treated with all together 20 single tooth implants were enrolled in the study. In 9 cases a second stage surgery after conventional submerged implant healing was performed, in the remaining 11 cases immediate implants were planned prior to tooth extraction. Beginning 1 day after implant installation and provisional restoration according to the copy abutment technique and 1 day after healing abutment insertion, respectively, peri-implant crevicular fluid samples (PICF) were collected, the PICF flow rate was recorded and laboratory assays performed for the assessment of interleukin-1β (IL-1β) and neutrophil elastase (NPE) concentrations (T1). Follow-up intervals were scheduled 1–3 weeks postoperatively (T2–4). Statistical analyses included explorative data analysis and median statistics (Wilcoxon test). Wound healing was uneventful in all cases and the concentrations of IL-1β and NPE remained stable over the entire observation period. Standard deviations and medians for IL-1β were higher in the second stage implant group with statistically significant differences in the concentrations on T2 and T4. No differences were found for NPE. The small sample size of this pilot investigation requires careful interpretation of the data presented. Within the limitations of this study it may be concluded that both immediately placed implants and implants at second stage surgery show similar patterns of wound healing with regard to selected markers of peri-implant disease, with minor differences in IL-1β concentrations on follow-up in favor of immediate implantation and restoration.

Ankylosed teeth are considered in orthodontic treatment planning; however, diagnostic tools to quantify the rigidity of the tooth-to-bone connection are rare. Resonance frequency analysis (RFA) can quantify the rigidity of the dental implant-to-bone connection and thus may serve as a potential diagnostic tool to identify ankylosed teeth. To test this assumption, we examined 15 and 30 primary mandibular molars, with and without clinical signs of ankylosis, using the Osstell Mentor system. A cut-off implant stability quotient (ISQ) of 43 provided a specificity of 100% and a sensitivity of 53.3% when measured in the mesio-distal direction or a sensitivity of 20% when measured in the bucco-lingual direction. Based on a receiver-operating characteristic (ROC), the area under the curve (AUC) of 0.807 showed the mesio-distal direction of measurement to be a test of moderate discriminatory power. Given its non-invasiveness, RFA may serve as a quantitative diagnostic supplement to the clinical examination of potentially ankylosed primary molars.

Bei thermisch gebogenen TVG- und ESG-Scheiben konnten in der Vergangenheit bei Schadensfallen und Forschungsprojekten durch Visualisierung mittels Polfilteraufnahmen Unregelmasigkeiten hinsichtlich der Verteilung der thermischen Vorspannung festgestellt werden. Die Abweichungen von einer homogenen Vorspannungsverteilung steht in Zusammenhang mit nicht konformen Bruchbildern [1]. Bis dato fehlte jedoch eine Interpretationsgrundlage fur die Bewertung der Polfilterbilder, da der Zusammenhang zwischen der visualisierten Farbgebung und dem Spannungszustand im Glas unbekannt war. Das Forschungsprojekt “ZIM – Gebogenes Glas“ nahm dies zum Anlass, die Moglichkeiten einer Online-Qualitatskontrolle mit Visualisierung der Vorspannungsverteilung zur Verbesserung der Produktqualitat von gebogenem Glas zu untersuchen und zerstorungsfrei Aussagen uber das Bruchbild zu treffen. Quality control of heat strengthened and fully tempered glass panels – Use of photoelasticity. In the past irregularities concerning the distribution of prestress in bent heat strengthened and fully tempered glass have been visualized by using polarizing filters. Especially in cases of damages, discrepancies of homogeneous distributions have been detected in polarizing filter images and a correlation between non-conforming fracture pattern has been made. Until now a basis for the interpretation of these irregularities could not be found. Therefore, within the research project “ZIM – bended glass” the product quality of bent glass by using the visualization of the distribution of stress was investigated. The results shall also serve for a future online quality monitoring system. The distribution of stress combined with the visualized colors gives a deeper insight in the resulting fracture patterns of glass.

Background: Autogenous bone grafts obtained by different harvesting techniques behave differently during the process of graft consolidation; the underlying reasons are however not fully understood. One theory is that harvesting techniques have an impact on the number and activity of the transplanted cells which contribute to the process of graft consolidation. Materials and Methods: To test this assumption, porcine bone grafts were harvested with four different surgical procedures: bone mill, piezosurgery, bone drilling (bone slurry), and bone scraper. After determining cell viability, the release of molecules affecting bone formation and resorption was assessed by reverse transcription polymerase chain reaction and immunoassay. The mitogenic and osteogenic activity of the conditioned media was evaluated in a bioassay with isolated bone cells. Results: Cell viability and the release of molecules affecting bone formation were higher in samples harvested by bone mill and bone scraper when compared with samples prepared by bone drilling and piezosurgery. The harvesting procedure also affected gene expression, for example, bone mill and bone scraper samples revealed significantly higher expression of growth factors such as bone morphogenetic protein-2 and vascular endothelial growth factor compared with the two other modalities. Receptor activator of nuclear factor kappa B ligand expression was lowest in bone scraper samples. Conclusion: These data can provide a scientific basis to better understand the impact of harvesting techniques on the number and activity of transplanted cells, which might contribute to the therapeutic outcome of the augmentation procedure.

Aim Vitamin D deficiency is highly prevalent in the population and associated with impaired peri-implant bone regeneration. Yet, there is a gap in understanding the impact of vitamin D supplementation on the process of osseointegration. In this study, the effect of vitamin D supplementation on peri-implant bone regeneration was investigated. Methods Fifty ovariectomized Sprague–Dawley rats were divided into three groups. The depletion group was fed a vitamin D-free diet for 8 weeks. The repletion group received vitamin D-free diet for 6 weeks, before animals were switched to standard diet containing 2400 IU/kg vitamin D. The control group was fed the standard diet. Two titanium mini-implants were placed in the tibia. All groups remained on their previous diet until sacrifice. Blood sample testing and histomorphometric analysis were performed. Results Vitamin D depletion caused a significant reduction in 25-hydroxvitamin D in rat serum that returned to control levels in the repletion group. This vitamin deficiency was associated with a decrease in bone-to-implant contact in the cortical area, which was leveled to controls in the repletion group. No significant changes by vitamin D depletion were noticed in the medullar compartment. Moreover, also the peri-implant bone area and the mineral apposition rate remained unchanged upon vitamin D depletion. Conclusion These results indicate that vitamin D deficiency has a negative impact on cortical peri-implant bone formation in ovariectomized rats, which can be compensated by vitamin D supplementation. This study provides first insight into the potential beneficial effect of vitamin D supplementation in implant dentistry.

Agis H, Watzek G, Gruber R. Prolyl hydroxylase inhibitors increase the production of vascular endothelial growth factor by periodontal fibroblasts. J Periodont Res 2012; 47: 165–173. © 2011 John Wiley & Sons A/S Background and Objective: Pharmacological inhibitors of prolyl hydroxylases (PHDs) can induce a proangiogenic response that favors wound healing and bone regeneration. However, the response of periodontal cells to PHD inhibitors is unknown. Material and Methods: To determine the effects of PHD inhibitors on periodontal cells, we exposed human fibroblasts from the gingiva and the periodontal ligament to dimethyloxallyl glycine, desferrioxamine, l-mimosine and CoCl2. Viability, proliferation, and protein synthesis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), [3H]thymidine, and [3H]leucine incorporation, respectively. The levels of Ki67, hypoxia-inducible factor 1α (HIF-1α), p27, phosphorylated c-Jun N-terminal kinase (JNK) and phosphorylated p38 were determined by immunohistochemistry and western blotting. Vascular endothelial growth factor (VEGF) mRNA levels were measured by quantitative PCR. Protein levels of VEGF and interleukin (IL)-6 were evaluated by immunoassays. Results: We found that PHD inhibitors, while leaving cell viability unchanged, reduced proliferation and protein synthesis. This was paralleled by decreased Ki67 levels and increased p27 levels, suggesting that PHD inhibitors provoke growth arrest. Independently from this response, PHD inhibitors stabilized HIF-1α and increased the production of VEGF. This increase of VEGF was observed in the presence of proinflammatory IL-1 and pharmacological inhibitors of JNK and p38 signaling. Moreover, PHD inhibitors did not modulate expression of IL-6 and the phosphorylation of JNK and p38. Conclusion: These results suggest that PHD inhibitors enhance the production of VEGF in periodontal fibroblasts, even in the presence of proinflammatory IL-1. The data further suggest that PHD inhibitors do not provoke a significant proinflammatory or anti-inflammatory response in this in vitro setting.

Dvorak G, Arnhart C, Heuberer S, Huber CD, Watzek G, Gruber R: Peri-implantitis and late implant failures in postmenopausal women: a cross-sectional study. J Clin Periodontol 2011; 38: 950–955. doi: 10.1111/j.1600-051X.2011.01772.x. Abstract Aim: Systemic bone loss is a major cause of fractures in postmenopausal women and may also affect the jawbone; however, its consequences on the success of dental implants remain poorly understood. Materials and Methods: In this cross-sectional study, the relation between self-reported osteoporosis and the success rate of dental implants in an adult female population was evaluated. The primary outcome parameters were the occurrence of peri-implantitis and late implant failures. Women with unknown bone status were excluded from the study. The potential confounders age, recipient site, smoking, periodontal disease and time of loading were recorded. Results: Data from 203 women with a mean age of 63±9 years and 967 dental implants were investigated. The patients were classified according to their medical history into one of three groups: osteoporosis (47 women), osteopenia (16 women) and healthy controls (140 women). Patients with unknown bone status (n=26) were excluded. The multi-level statistical analysis showed no association between peri-implantitis [odds ratio (OR) 2.1; p=0.6] or implant failure [hazards ratio (HR) 2.5; p=0.2] and systemic bone loss. Conclusions: No relation was found between osteoporosis and peri-implantitis in an adult female population.

The bond joint between dentin and ceramic is a critical determinant in prosthodontic dentistry. The laser is an alternative to the diamond bur for preparing tooth cavities. However, the impact of lasers on the bond between the laser-irradiated dentin and the ceramic remains a matter of controversy. We determined the shear strength of bonds between ceramic blocks and human dentin discs prepared with either an Er,Cr:YSGG laser or a diamond bur. A total of 180 dentin discs were randomly assigned to four groups. Three groups of discs were prepared with the Er,Cr:YSGG laser irradiation (2 W, 30 Hz, 50% H(2)O, 70% air) and the fourth group was prepared with a diamond bur. In one of the laser groups the discs surfaces were also treated with phosphoric acid and in another with phosphoric acid and mechanical smoothing using a dental excavator. The ceramic blocks were bonded to the dentin discs with Syntac adhesive (together with Variolink II curing system), ExciTE adhesive (together with Variolink II curing system) or RelyX self-adhesive cement. The shear strength of the bond between ceramic and dentin was significantly higher following dentin surface treatment with the laser alone than following treatment with the diamond bur and Variolink II/Syntac (p = 0.021) but not significantly higher than following treatment with the diamond bur and Variolink II/ExciTE (p = 0.138) or RelyX (p = 0.150). A significant difference was not observed when the laser-treated dentin was conditioned with phosphoric acid and mechanical smoothing. These findings demonstrate that the bond between dentin and ceramic may be stronger after laser irradiation; however, the selection of the adhesive material is an additional factor that affects the bond strength.

Lipoxins and resolvins are endogenous eicosanoids which participate in the resolution of inflammation. The connective tissue, including the fibroblasts of the periodontal ligament and the gingiva, is inflamed in periodontal disease. However, it is unknown if periodontal fibroblasts are targeted by lipoxins and resolvins. This study has investigated the impacts of lipoxin A4, lipoxin B4, and resolvin D1 on the response of periodontal ligament fibroblasts and gingival fibroblasts to the inflammatory mediators interleukin (IL)-1b and tumor necrosis factor-a (TNF-a) and platelet-derived growth factor BB (PDGF-BB). It was found that up to a concentration of 1 mM neither of the eicosanoids changed the expression of IL-6 and IL-8 by periodontal fibroblasts previously exposed to IL-1b and TNF-a. The eicosanoids also failed to modulate IL-1b-induced phosphorylation of p38 and c-Jun N-terminal protein kinase signaling molecules. Furthermore, lipoxins and resolvin D1 did not change proliferation rates or protein synthesis by periodontal fibroblasts exposed to PDGF-BB. Together, these results suggest that the response of periodontal fibroblasts to inflammatory mediators and a potent mitogen is not modulated by lipoxins and resolvin D1 under the in vitro conditions studied.

Teriparatide is an anabolic osteoporosis therapeutic agent that can improve healing after fractures and periodontal surgeries. Clinical studies investigating the effects of teriparatide on the osseointegration of titanium implants have not been performed. We conducted an open-label randomized controlled feasibility study and included 24 individuals with edentulous lower jaws. The participants received 2 study implants in the mandible during interforaminal dental implant surgery. They were randomly assigned to receive either 20 µg of teriparatide once daily for 28 days or no treatment. Study implants were retrieved from 23 participants after 9 weeks and were subjected to histomorphometric analyses. Endpoints were new bone-volume-per-tissue-volume (NBV/TV) and new bone-to-implant-contact (NBIC). We report here that median values of NBV/TV in the control and the teriparatide groups were 15.4% vs. 17.6% in the periosteal compartment, 11.3% vs. 16.5% in the cortical compartment, and 7.3% vs. 12.0% in the medullary compartment, respectively. NBIC median values in the control and the teriparatide groups were 3.3% vs. 4.1% in the periosteal compartment, 5.0% vs. 4.4% in the cortical compartment, and 0.3% vs. 1.4% in the medullary compartment, respectively. The results provide the first histological data on the osseointegration of titanium study implants in individuals treated with teriparatide. ClinicalTrials.gov number, NCT00089674. Abbreviations: NBV/ TV, new bone-volume per tissue volume; NBIC, new bone-to-implant contact; BV/TV, bone-volume per tissue volume; BIC, bone-to-implant contact; ROI, region of interest.