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BACKGROUND/PURPOSE Dalbergia cochinchinensi has been widely used in traditional medicine because of its flavonoids. This study examined which components in D. cochinchinensis were capable of reducing or even stimulating the formation of bone-resorbing osteoclasts. MATERIALS AND METHODS We have isolated subfamilies of chalcones (isoliquiritigenin, butein), flavones (7-hydroxy-6-methoxyflavone) and neoflavanoids (5-methoxylatifolin), and performed an in vitro bioassay on osteoclastogenesis. The flavonoids were tested for their potential to change the expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin K (CTSK) in murine bone marrow cultures being exposed to RANKL, M-CSF and TGF-β1 using RT-PCR, histochemistry and immunoassay. RESULTS We could confirm that isoliquiritigenin and butein significantly lower the expression of TRAP and CTSK in this setting. Moreover, histochemistry supported the decrease of TRAP by the chalcones. We further observed a trend towards an increase of osteoclastogenesis in the presence of 5-methoxylatifolin and 7-hydroxy-6-methoxyflavone, particular in bone marrow cultures being exposed to RANKL and M-CSF. Consistently, the anti-inflammatory activity was restricted to isoliquiritigenin and butein in murine RAW 264.7 inflammatory macrophages stimulated by lipopolysaccharide (LPS). With respect to osteoblastogenesis, neither of the flavonoids but butyrate, a short chain fatty acid, increased the osteogenic differentiation marker alkaline phosphatase activity in ST2 murine mesenchymal cells. CONCLUSION We have identified two flavonoids from D. cochinchinensis with a potential pro-osteoclastogenic activity and confirm the anti-osteoclastogenic activity of isoliquiritigenin and butein.

It has been proposed that platelet-rich fibrin (PRF) can be used to support bone regeneration during alveolar ridge augmentation. The aim of this study was to determine whether an approach utilizing PRF provides similar performance to the established guided bone regeneration (GBR) procedure.Two-wall defects were surgically created in beagle dogs and treated in three experimental groups: (i) a sticky bone (SB) substitute prepared using liquid PRF and deproteinized porcine bone mineral (DPBM); (ii) SB covered with solid PRF compressed into a membrane; and (iii) GBR performed using DPBM covered by a collagen membrane. Quantitative reverse-transcription polymerase chain reaction was applied to the specimen after 1 week of healing, and microcomputed tomography (micro-CT) and histological outcomes were analysed after 8 weeks of healing.Compared with GBR, PRF resulted in a moderate increase in the expression levels of osteoblast and osteoclast markers, osteocalcin, and calcitonin receptor. Moreover, PRF modestly increased angiogenesis and the inflammation markers vascular endothelial growth factor (VEGF) and IL-6. Micro-CT and histological analyses confirmed the expected increased alveolar ridge area, with no significant differences between the three groups. Consistently, graft consolidation, as indicated by new bone formation at the defect site, did not differ significantly between groups.The present results demonstrate that PRF-based approaches perform comparably to the established GBR procedure in terms of the consolidation of DPBM in two-wall alveolar defects.

Fas ligand (FasL) belongs to the tumour necrosis factor superfamily regulating bone turnover, inflammation, and apoptosis. The appendicular and axial skeleton phenotype of mature FaslWe performed a phenotypical analysis of mice carrying the homozygous FaslHealthy FaslOur findings suggest that FasL is a catabolic factor in alveolar bone homeostasis but it does not affect the inflammatory osteolysis.

Background Secretomes of mesenchymal stromal cells (MSC) are emerging as a novel strategy for growth factor delivery and a promising alternative to cell therapies for tissue regeneration. The objective of this study was to compare the efficacy of adjunctive use of conditioned media of human MSC (MSC-CM) with collagen barrier membranes (MEM) for guided bone regeneration (GBR) vs. adjunctive use of conditioned media of leukocyte- and platelet-rich fibrin (PRF-CM).Methods MSC-CM and PRF-CM were prepared from three human donors each and subjected to quantitative proteomic analysis using mass spectrometry and multiplex immunoassay. Early responses (48 h) of MSC to MSC-CM and PRF-CM were studied via gene expression analysis. Next, MEM functionalized with pooled MSC-CM or PRF-CM were applied on critical-size rat calvaria defects and new bone formation was analyzed via micro-computed tomography (µCT, 2 and 4 weeks) and histology (4 weeks).Results Proteomic analysis revealed several proteins representing biological processes related to bone formation in both MSC-CM and PRF-CM, although more differentially expressed proteins (p Conclusion Based on the proteomic analysis and in vivo investigation, MSC-CM functionalized on MEM represents a promising strategy to enhance GBR.

Secretomes of mesenchymal stromal cells (MSC) are emerging as a novel growth factor (GF)-based strategy for periodontal and bone regeneration. The objective of this study was to compare the secretome of human bone marrow MSC (BMSC) to that of leukocyte- and platelet-rich fibrin (L-PRF), an established GF-based therapy, in the context of wound healing and regeneration. Conditioned media from human BMSC (BMSC-CM) and L-PRF (LPRF-CM) were subjected to quantitative proteomic analysis using liquid chromatography with tandem mass spectrometry. Global profiles, gene ontology (GO) categories, differentially expressed proteins (DEPs), and gene set enrichment (GSEA) were identified using bioinformatic methods. Concentrations of selected proteins were determined using a multiplex immunoassay. Among the proteins identified in BMSC-CM (2157 proteins) and LPRF-CM (1420 proteins), 1283 proteins were common. GO analysis revealed similarities between the groups in terms of biological processes (cellular organization, protein metabolism) and molecular functions (cellular-/protein-binding). Notably more DEPs were identified in BMSC-CM (n=550) compared to LPRF-CM (n=118); these included several key GF, cytokines, and extracellular matrix (ECM) proteins involved in wound healing. GSEA revealed enrichment of ECM (especially bone-ECM) related processes in BMSC-CM and immune-related processes in LPRF-CM. Similar trends for inter-group differences in protein detection were observed in the multiplex analysis. Thus, while the secretomes of both BMSC and L-PRF are favourable for wound healing, BMSC-CM is additionally enriched for proteins/processes relevant for periodontal and bone regeneration.

OBJECTIVES Resonance frequency analysis (RFA) is used to monitor implant stability. Its output, the Implant Stability Quotient (ISQ), supposedly correlates with insertion torque, a common measurement of primary stability. However, the reliability of RFA in condensed bone remains unclear. MATERIAL AND METHODS In this human cadaver study in edentulous jaws and fresh extraction sockets, implants were inserted using a split-mouth approach into condensed or untreated bone. Mean ISQ, peak insertion torque, and pre- and postoperative bone volume fractions (BV/TV) were assessed. RESULTS In edentulous jaws, insertion torque and ISQ correlated both in untreated (r��=��0.63, p��=��0.02) and in condensed (r��=��0.82, p����< 0.01) bone. In extraction sockets, insertion torque and ISQ only correlated in untreated (r��=��0.78, p��

Objectives: to study how cellular lysates could affect the expression of inflammatory cytokines of periodontal-related cells. Materials and Methods: cell suspensions were submitted to different treatments: lysate 1 – no treatment; lysate 2 – sonication; and lysate 3 – freeze/thawing. All lysates were centrifuged, and the supernatants were used for experimentation. Cell viability assays, RT-qPCR of IL1, IL6 and IL8, IL6 immunoassay, and immunofluorescence of NF-kB were applied to verify the inflammatory crosstalk potential of damaged cells over healthy plated cells. Further, titanium discs and collagen membranes were treated with the cell lysates and checked for IL8 RT-qPCR expression. Results: lysates obtained upon sonication or freeze/thawing of oral squamous cell carcinoma cell lines provoked a robust increase in the expression of IL1, IL6, and IL8 by gingival fibroblasts, which was confirmed by immunoassays for IL6. Lysates obtained from the gingival fibroblasts failed to increase the expression of inflammatory cytokines in oral squamous cell carcinoma cells. Epithelial cell lysates caused the activation of the NF-kB signalling cascade in gingival fibroblasts as indicated by the phosphorylation and nuclear translocation of p65, indicating inflammation. Finally, epithelial lysates could adhere to the titanium and collagen membrane surfaces and increase IL8 expression by gingival fibroblasts. Conclusions: injured oral epithelial cells can release factors that incite gingival fibroblasts to become pro-inflammatory. Clinical Relevance: injuries affecting the oral mucosa generate epithelial fragments that may reach the underlying connective tissue and provoke inflammation. These injuries are routinely caused by mastication, sonication for teeth cleaning, prostheses maladaptation, implant drilling, and teeth preparation.

Objective To investigate the efficacy of platelet-rich fibrin (PRF) for the socket healing after removal of mandibular third molars when compared to natural wound healing. Methods This systematic review and meta-analysis were conducted according to PRISMA guidelines. The eligibility criteria comprised randomized controlled trials (RCTs) comparing the clinical outcomes of PRF with that of natural healing. Outcomes measured include post-operative pain, analgesics taken, swelling, soft tissue healing, bone healing, complication rates, as well as incidence of alveolar osteitis (AO, dry socket). Results From 194 articles identified, 18 RCTs were included. Overall, 15 of 18 RCTs demonstrated significantly better outcomes in terms of reduced complication rates, healing rates, or patient reported pain scores in the PRF group when compared to control. Data from all studies demonstrated that PRF was able to significantly lower the rate of AO from 15.9 % in the control group to 5.8 % in the PRF group, representing a ∼3-fold reduction. All studied investigating soft tissue healing with PRF demonstrated better outcomes in favor of PRF group. Conclusions PRF was shown to decrease post-op pain/morbidity, have favored soft tissue healing, and possess lower complication rates or AO incidence. PRF therefore proves to be a viable treatment adjunct following removal of mandibular third molars.

Aim To determine the healing outcome following grafting with deproteinized porcine bone mineral (DPBM) with or without collagen membrane coverage in two‐wall (both buccal and lingual)‐damaged extraction sockets. Materials and methods Distal roots of three mandibular premolars in six beagle dogs were extracted, and the whole buccal and lingual bony walls were surgically removed. Three treatment protocols were then applied according to the following group allocation: no graft (None), grafting DPBM (BG), and grafting DPBM with coverage by a collagen membrane (BG + M). Two observational periods (2 and 8 weeks) were used with the split‐mouth design, and quantitative and qualitative analyses were performed by microcomputed tomography and histology. Results The dimensions of the alveolar ridge at both grafted sites (BG and BG + M) remained similar to those of the pristine ridge in the histologic and radiographic analyses, whereas the ungrafted sites (None) collapsed both vertically and horizontally. Both grafting protocols produced substantial bony regeneration, but the addition of a covering membrane enhanced the proportion of mineralized tissue within the augmented area, and the BG + M group also showed a significantly larger area of regenerated ridge than the None group (p

Objectives To investigate how scaling affects the penetration of microorganisms into dentinal tubules, how pulpal cells seeded into the pulp cavity respond to bacterial challenge, and how penetration and inflammatory response may depend on the bacterial composition. Materials and methods Root canals of 102 extracted human teeth underwent shaping and cleaning. Half of the teeth were subjected to scaling and root planing, the other half remained untreated. Teeth were exposed to either Streptococcus gordonii and Actinomyces oris or S. gordonii and Porphyromonas gingivalis for 10 weeks. Bacterial invasion was assessed in a depth of 1 mm to the root surface. Human pulpal cells were seeded into the cavities to assess the expression of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and matrix metalloproteinase-3 (MMP-3) by real-time polymerase chain reaction and immunoassay. Results The percentage of teeth with bacteria detected in dentine was higher when teeth received scaling than when they were untreated: 66.6% versus 44.4% when exposed to A. oris/S. gordonii, and 50% versus 25% when exposed to P. gingivalis/S. gordonii (p = 0.043). Scaling had no impact on IL-8 and MMP-3 expression in pulpal cells. P. gingivalis/S. gordonii caused higher levels of IL-8, MCP-1, and MMP-3 than A. oris/S. gordonii (p = 0.003, p = 0.011, p = 0.037). Conclusion Scaling supports the penetration of bacteria into the dentine of extracted human teeth. P. gingivalis may affect the immune response in pulpal cells. Clinical relevance Root surface debridement with hand instruments may facilitate bacterial penetration. Other kinds of mechanical instrumentation in this experimental setting should be investigated.

Background The present study focused on the inflammatory disease progress after periodontal defect induction and aimed to specifically determine periodontal tissue responses following dental plaque accumulation by ligatures on a site with/without standardized periodontal defect induction. Methods After 1 month from extraction of the adjacent teeth, semi-circumferential defects were surgically created in the unilateral second and fourth premolars (test group), whereas no defects were being induced at the contralateral sites (control group). One week later, silk was used to ligate the tooth cervix at both sites to encourage the accumulation of dental plaque. Four weeks later, the tissue samples were collected for histological/histomorphometric and microarray analysis. Microbiological analysis was performed before defect induction and at ligatures, and after 4 weeks of dental plaque accumulation. Results Remarkable inflammation was clinically and histologically observed in both groups after plaque accumulation, and the intrabony type of periodontal defect exaggerated inflammatory cell infiltration into the connective tissue layer. Expression of genes related to inflammation such as IL-1 was highly up-regulated in test sites. However, these inflammatory infiltrations did not invade to a boundary of periodontal ligament and connective tissue attachment in both groups, and histomorphometric results corresponds to these observational results. Bacterial findings also showed no significant differences in detected microbiome compositions between control and test groups at three-time points. Conclusion Intrabony defect might exaggerate the plaque-induced inflammation in the aspect of inflammatory cell infiltration and the related gene expression, but both dental plaque and the pre-existing periodontal defect negligibly disrupt periodontal attachment and the underlying alveolar bone.

OBJECTIVE The aim of the study was to determine whether the inhibition of apoptosis via pan-caspase inhibitors can attenuate the changes in the alveolar ridge upon tooth extraction. BACKGROUND Cells undergoing apoptosis might play a central role in the onset of alveolar bone resorption and the ensuing bone atrophy following tooth extraction. Caspases are proteases that regulate apoptotic cell death. It is, therefore, reasonable to hypothesize that blocking apoptosis with pan-caspase inhibitors attenuates the changes in the alveolar ridge following tooth extraction. METHODS In 16 inbred rats, the mandibular first (M1) and second (M2) molars of one side were extracted. Following random allocation, the rats received either a cell-permeable pan-caspase inhibitor or diluent. After a healing period of 10��days, changes in shape and height of the alveolar ridge were examined using geometric morphometrics and linear measurements based on micro-computed tomography. RESULTS Geometric morphometric analysis revealed that the pan-caspase inhibitor prevented major shape changes of the alveolar ridge following M1 tooth extraction (P��

Objective: Iliac crest autografts can compensate for severe mandibular atrophy before implant placement. However, the implant success in the augmented bone is not entirely predictable. Here we performed a retrospective cohort study to determine the success and related parameters of implants placed in augmented bone and pristine bone for up to 11 years. Material and Methods: We analyzed 18 patients where 72 implants were placed six months after iliac crest transplantation and 19 patients where 76 implants were placed in pristine bone. The primary endpoint was implant loss. Secondary endpoints were the implant success, peri-implant bone loss, and the clinical parameters related to peri-implantitis. Moreover, we evaluated the oral-health-related quality of life (OHIP). Results: Within a mean follow-up of 5.8 ± 2.2 and 7.6 ± 2.8 years, six but no implants were lost when placed in augmented and pristine bone, respectively. Among those implants remaining in situ, 58% and 68% were rated as implant success (p = 0.09). A total of 11% and 16% of the implants placed in the augmented and the pristine bone were identified as peri-implantitis (p = 0.08). Bone loss was similar in both groups, with a mean of 2.95 ± 1.72 mm and 2.44 ± 0.76 mm. The mean OHIP scores were 16.36 ± 13.76 and 8.78 ± 7.21 in the augmentation and the control group, respectively (p = 0.35). Conclusions: Implants placed in iliac crest autografts have a higher risk for implant loss and lower implant success rates compared to those placed in the pristine bone.

Liquid platelet-rich fibrin (PRF) is produced by fractionation of blood without additives that initiate coagulation. Even though liquid PRF is frequently utilized as a natural source of fibrinogen to prepare sticky bone, the concentration of fibrinogen and the overall amount of “clottable PRF” components have not been evaluated. To this aim, we prepared liquid PRF at 300, 700, and 2000 relative centrifugal force (RCF), for 8 min and quantified the fibrinogen levels by immunoassay. We report here that, independent of the RCF, the fibrinogen concentration is higher in the platelet-poor plasma (PPP) compared to the buffy coat (BC) fraction of liquid PRF and further decreases in the remaining red fraction. We then determined the weight of the clotted PRF fractions before and after removing the serum. The PPP and BC fractions consist of 10.2% and 25.3% clottable matrix suggesting that more than half of the weight of clottable BC is caused by cellular components. Our data provide insights into the distribution of fibrinogen in the different fractions of liquid PRF. These findings suggest that PPP is the main source of clottable fibrinogen, while the BC is more a cell source when it comes to the preparation of sticky bone.

Background: Pyroptosis is a caspase-dependent catabolic process relevant to periodontal disorders for which inflammation is central to the pathophysiology of the disease. Although enamel matrix derivative (EMD) has been applied to support periodontal regeneration, its capacity to modulate the expression of pyroptosis-related genes remains unknown. Considering EMD has anti-inflammatory properties and pyroptosis is linked to the activation of the inflammasome in chronic periodontitis, the question arises whether EMD could reduce pyroptosis signalling. Methods: To answer this question, primary macrophages obtained from murine bone marrow and RAW 264.7 macrophages were primed with EMD before being challenged by lipopolysaccharide (LPS). Cells were then analysed for pyroptosis-signalling components by gene expression analyses, interleukin-1β (IL-1β) immunoassay, and the detection of caspase-1 (CAS1). The release of mitochondrial reactive oxygen species (ROS) was also detected. Results: We report here that EMD, like the inflammasome (NLRP3) and CAS1 specific inhibitors—MCC950 and Ac-YVAD-cmk, respectively—lowered the LPS-induced expression of NLRP3 in primary macrophages (EMD: p = 0.0232; MCC950: p = 0.0426; Ac-YVAD-cmk: p = 0.0317). EMD further reduced the LPS-induced expression of NLRP3 in RAW 264.7 cells (p = 0.0043). There was also a reduction in CAS1 and IL-1β in RAW 264.7 macrophages on the transcriptional level (p = 0.0598; p = 0.0283; respectively), in IL-1β protein release (p = 0.0313), and CAS1 activity. Consistently, EMD, like MCC950 and Ac-YVAD-cmk, diminished the ROS release in activated RAW 264.7 cells. In ST2 murine mesenchymal cells, EMD could not be tested because LPS, saliva, and IL-1β + TNF-α failed to provoke pyroptosis signalling. Conclusion: These findings suggest that EMD is capable of dampening the expression of pyroptosis-related genes in macrophages.

Platelet-rich fibrin (PRF) has a potent anti-inflammatory activity but the components mediating this effect remain unknown. Blood lipids have anti-inflammatory properties. The question arises whether this is also true for the lipid fraction of PRF. To answer this question, lipid fractions of solid and liquid PRF were tested for their potential to lower the inflammatory response of ST2 bone marrow stromal cells and primary bone marrow macrophages exposed to IL1β and TNFα, and LPS, respectively. Cytokine production and the underlying signalling pathway were analysed by RT-PCR, immunoassays, and Western blotting. We report here that lipids from solid and liquid PRF substantially lowered cytokine-induced expression of IL6, CCL2 and CCL5 in ST2 cells. Moreover, the inflammatory response induced by Pam3CSK4, the agonist of Toll-like receptor (TLR) TLR2, was partially reduced by the lipid extracts in ST2 cells. The PRF lipids further reduced the LPS-induced expression of IL1β, IL6 and CCL5 in macrophages at the transcriptional level. This was confirmed by showing the ability of PRF lipids to diminish IL6 at the protein level in ST2 cells and macrophages. Likewise, PRF lipid extracts reduced the phosphorylation of p38 and JNK and moderately decreased the phosphorylation of NFκB-p65 in ST2 cells. These findings suggest that the lipid fraction is at least partially responsible for the anti-inflammatory activity of PRF in vitro.

Digital technology facilitates the manufacturing of complete dentures; however, clinical and patient-reported outcomes are underreported. This prospective, randomized, single-blind cross-over study reports the clinical and patient-related outcomes of 10 edentulous patients receiving digital dentures prepared with the Vita Vionic System and conventional dentures produced from heat-polymerized polymethylmethacrylate resin. Clinical efficiency was stated based on the Sato score for quantitative assessment of complete denture quality. Patient satisfaction was evaluated with the oral health-related quality of life questionnaire (OHIP-20). We report here that the Sato score was slightly higher in patients receiving digital versus conventional dentures with a mean of 73.2 ± 12.3 and 67.4 ± 11.8, respectively (p = 0.16). Moreover, upper and lower stability was superior in digital dentures (p = 0.03 and p = 0.10, respectively), while denture polish was better in conventional dentures (p = 0.03). Quality of life was slightly higher in patients receiving conventional compared to digital dentures with an OHIP-20 of 101.7 ± 12.0 and 95.6 ± 24.2, respectively (p = 0.33). Taken together and when considering the low power of the study, our findings suggest a trend towards better clinical efficiency of digital compared to conventional dentures, while patient satisfaction remained unaffected by the type of manufacturing.

Taurocholic acid (TCA), a conjugation of cholic acid with taurine, is one of the main bile acids that is elevated in liver disease. Considering the epidemiologic linkage of periodontal disease to liver disease, the question arises about the possible effect of elevated TCA levels on periodontal cells. To answer this question, gingival fibroblasts and human oral squamous cell carcinoma cell line (HSC-2) were pretreated with interleukine1β (IL1β) and tumor necrosis factorα (TNFα) in the presence and absence of TCA. Also, mouse macrophages (RAW 264.7) were incubated with sterile-filtered human saliva with and without TCA. Inflammatory cytokines were measured by real time polynucleotide chain reaction (RT-PCR) and an immunoassay. The nuclear translocation of the p65 subunit was visualized by immunostaining. In pretreated gingival fibroblasts and HSC-2 cells, TCA considerably reduced the expression of IL1β, IL6, and IL8. In support of these observations, TCA lowered the saliva-induced expression of IL1α, IL1β and IL6 in RAW 264.7 cells. An immunoassay confirmed the capacity of TCA to diminish inflammation-induced expression of IL6 in gingival fibroblasts, HSC-2 and RAW 264.7 cells. Consistently, TCA blocked the nuclear translocation of p65 in fibroblasts. These findings suggest that TCA has anti-inflammatory activity in gingival fibroblasts, human oral squamous cell carcinoma cells and macrophages in vitro.

Sclerosteosis, a rare autosomal recessive genetic disorder caused by a mutation of the Sost gene, manifests in the facial skeleton by gigantism, facial distortion, mandibular prognathism, cranial nerve palsy, and, in extreme cases, compression of the medulla oblongata. Mice lacking sclerostin reflect some symptoms of sclerosteosis, but this is the first report of the effect on the facial skeleton. We used geometric morphometrics (GMM) to analyze the deformations of the murine facial skeleton from the wild‐type to the Sost gene knockout. Landmark coordinates were obtained by surface reconstructions from micro‐computed tomography. Centroid size, principal component scores in shape space and form space, and asymmetry were computed by the standard GMM formulas, and dental and skeletal jaw lengths were examined as ratios. We show here that, compared to wild type controls, mice lacking Sost have larger centroid size (effect size, p‐value: 4.59

Aim Our objective was to compare guided virtual surgery to conventional surgery in terms of angular deviation of single dental implants placed in the posterior mandible. Materials and methods Patients with bilateral homologous single teeth missing in the posterior mandible were eligible for this split-mouth randomized clinical trial. Cone beam computed tomography (CBCT) was performed for virtual planning of implant position and manufacturing of the stereolithographic guides. One week after the surgery, a second CBCT scan was superimposed to the initial planning. Primary endpoint was the angular deviation between virtual and clinical implant position. Secondary endpoints were linear deviations and patient-reported outcomes collected with a questionnaire. Results Data from 12 patients were available for analysis. Angular deviation was significantly lower using stereolithographic guides as compared to conventional guides (2.2 ± 1.1° vs. 3.5 ± 1.6°, p = .042). Linear deviations were similar for both techniques in the coronal (2.34 ± 1.01 vs. 1.93 ± 0.95 mm) and apical (2.53 ± 1.11 vs. 2.19 ± 1.00 mm) dimensions (p ˃ .05). The selection of the surgical technique had no significant impact on the patient-reported outcomes. Conclusion Our data suggest that the angular discrepancy between the virtual and the clinical implant position is slightly lower when using stereolithographic guides as compared to conventional guides.

Background The periodontal ligament (PDL) comprises cells embedded in the extracellular matrix (ECM) and is required for periodontal healing upon tooth transplantation. However, the extent to which the ECM contributes to periodontal regeneration remains unknown. This study aimed to evaluate effects of separate PDL matrix without cells on periodontal healing. Methods Mandibular premolars extracted from six beagle dogs, were 1) left untreated, 2) decellularized, or 3) surface planed. Cytologic, histologic, scanning electron microscopy, and transmission electron microscopy analyses confirmed that detergents could be used to decellularize the PDL. An additional circumferential osteotomy was performed to replant dental roots into extraction sockets. Radiography and histology were used to evaluate periodontal regeneration 8 weeks later, with the data adjusted for multiple testing. Results In pristine extraction sites, total root resorption (P = 0.034), recovered PDL space (P = 0.012), and new cementum (P = 0.004) were greater in untreated teeth than in roots that underwent surface planing. There were no significant changes when comparing untreated teeth with teeth having a decellularized PDL (P = 0.081, P = 0.170, and P = 1.000, respectively), and decellularized teeth showed significant increase of new cementum compared to surface planed teeth (P = 0.048). In the defect area, only the recovered PDL space (P = 0.034) was greater in untreated teeth when comparing with denuded roots. Conclusion These results suggest that in addition to untreated PDL, decellularized PDL also partially supports reattachment (particularly cementogenesis) in pristine extraction sites but not in defect areas.

Aim Osteoimmunology covers the cellular and molecular mechanisms responsible for inflammatory osteolysis that culminates in the degradation of alveolar bone. Osteoimmunology also focuses on the interplay of immune cells with bone cells during bone remodelling and regeneration. The aim of this review was to provide insights into how osteoimmunology affects alveolar bone health and disease. Method This review is based on a narrative approach to assemble mouse models that provide insights into the cellular and molecular mechanisms causing inflammatory osteolysis and on the impact of immune cells on alveolar bone regeneration. Results Mouse models have revealed the molecular pathways by which microbial and other factors activate immune cells that initiate an inflammatory response. The inflammation-induced alveolar bone loss occurs with the concomitant suppression of bone formation. Mouse models also showed that immune cells contribute to the resolution of inflammation and bone regeneration, even though studies with a focus on alveolar socket healing are rare. Conclusions Considering that osteoimmunology is evolutionarily conserved, osteolysis removes the cause of inflammation by provoking tooth loss. The impact of immune cells on bone regeneration is presumably a way to reinitiate the developmental mechanisms of intramembranous and endochondral bone formation.

Background and Aims: To describe the biology of alveolar bone regeneration.Material and Methods: Four comprehensive reviews were performed on (a) mesenchymal cells and differentiation factors leadi ...

Simvastatin embedded into poly(lactic-co-glycolic acid) (PLGA)-based scaffolds can stimulate bone regeneration in preclinical models. However, the ideal pharmacological dose has not been evaluated. This systematic review reports on the simvastatin doses used in preclinical studies and evaluates the regeneration of critical-sized bone defects. References were selected in a two-phase process. Electronic databases (Embase, LILACS, LIVIVO, PubMed, SCOPUS, and Web of Science) and grey literature databases (Google Scholar, Open Grey, and ProQuest) were searched until September 2022. The risk of bias was considered to be low based on the SYRCLE tool. We identified four studies in rat, two in parietal and two in calvaria bone, one in mouse parietal bone, and one in rabbit femur bone. Simvastatin, ranging from 8 to 100 µg, significantly increased bone formation in five studies, as compared to the scaffold alone based on µ-computed tomography, histomorphometric, and radiography analysis. The median increase in bone formation caused by simvastatin was 2.1-fold compared to the PLGA-based scaffold alone. There was, however, no significant correlation between the relative bone gain and the doses of simvastatin (p = 0.37). The data suggest that relatively lower doses of simvastatin can consistently promote preclinical bone regeneration. However, the interpretation of these data must consider the heterogenicity of the PLGA-scaffolds, the defect anatomy, the observation period, and the evaluation method.

To investigate the effect of platelet-rich fibrin on bone formation by investigating its use in guided bone regeneration, sinus elevation and implant therapy.This systematic review and meta-analysis were conducted and reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. The eligibility criteria comprised human controlled clinical trials comparing the clinical outcomes of platelet-rich fibrin with those of other treatment modalities. The outcomes measured included percentage of new bone formation, percentage of residual bone graft, implant survival rate, change in bone dimension (horizontal and vertical), and implant stability quotient values.From 320 articles identified, 18 studies were included. Owing to the heterogeneity of the investigated parameters, a meta-analysis was only possible for sinus elevation. There is a general lack of data from comparative randomised clinical trials evaluating platelet-rich fibrin for guided bone regeneration procedures (only two studies), with no quantifiable advantages in terms of new bone formation or dimensional bone gain found in the platelet-rich fibrin group. For sinus elevation, the meta-analysis demonstrated no advantage in terms of histological new bone formation in the control group (bone graft alone) compared with the test group (bone graft and platelet-rich fibrin). Two studies demonstrated that platelet-rich fibrin may shorten healing periods prior to implant placement. Platelet-rich fibrin was also shown to slightly enhance primary implant stability (implant stability quotient value5) as assessed using implant stability quotients and resonance frequency analysis parameters, with no histological data evaluating bone-implant contact yet available on this topic. In one study, platelet-rich fibrin was shown to improve the clinical parameters when utilised as an adjunct for the treatment of peri-implantitis.In the majority of studies, platelet-rich fibrin offered little or no clear advantage in terms of new bone formation as evaluated in various studies on guided bone regeneration and sinus elevation, nor in implant stability and treatment of peri-implantitis. Various authors and systematic reviews on the topic have now expressed criticism of the various study designs and protocols, and the lack of appropriate controls and available information regarding patient selection. Well-controlled human studies on these specific topics are required.

To investigate the use of platelet-rich fibrin for alveolar ridge preservation compared to natural healing, bone graft material and platelet-rich fibrin in combination with bone graft material.The present systematic review was conducted and reported according to the Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines. The review examined randomised controlled trials comparing the clinical outcomes of platelet-rich fibrin with those of other modalities for alveolar ridge preservation. Studies of third molar extraction site healing were excluded. The studies were classified into three categories: natural wound healing vs platelet-rich fibrin; bone graft material vs platelet-rich fibrin; and bone graft material vs bone graft material and platelet-rich fibrin.From 179 articles identified, 16 randomised controlled trials were included. Owing to the heterogeneity of the investigated parameters, it was not possible to perform a meta-analysis. In total, 10 randomised controlled trials compared platelet-rich fibrin to natural wound healing, with seven of these demonstrating favourable outcomes to either limit postextraction dimensional changes or improve new bone formation in the platelet-rich fibrin group. Three of four studies comparing healing with bone graft material to platelet-rich fibrin found that the latter led to significantly greater horizontal or vertical bone resorption, and the bone graft material was more able to maintain the ridge dimensions. Two out of three randomised controlled trials investigating healing with both bone graft material and platelet-rich fibrin reported better outcomes using this combined approach than with bone graft material alone. All studies investigating soft tissue healing with platelet-rich fibrin demonstrated better outcomes in the platelet-rich fibrin group.The majority of studies comparing healing with platelet-rich fibrin to natural healing concluded that the former more successfully limits postextraction dimensional changes than the latter. However, 75% of studies investigating platelet-rich fibrin vs bone graft material reported better results in the bone graft group with respect to its ability to maintain postextraction dimensional changes. The addition of platelet-rich fibrin to bone graft material may improve clinical outcomes, although data are limited.

Fas ligand (FasL) is a member of the tumor necrosis factor (TNF) superfamily involved in the activation of apoptosis. Assuming that apoptosis is initiated after tooth extraction it is reasonable to suggest that FasL may play a pivotal role in the healing of extraction sockets. Herein, we tested the hypothesis of whether the lack of FasL impairs the healing of extraction sockets. To this end, we extracted upper right incisors of FasL knockout (KO) mice and their wildtype (WT) littermates. After a healing period of two weeks, bone volume over total volume (BV/TV) via µCT and descriptive histological analyses were performed. µCT revealed that BV/TV in the coronal region of the socket amounted to 39.4% in WT and 21.8% in KO, with a significant difference between the groups (p=0.002). Likewise, in the middle region of the socket, BV/TV amounted to 50.3% in WT and 40.8% in KO (p

OBJECTIVES The extraction of impacted mandibular third molars is a common surgical procedure often associated with complications including post-traumatic neuropathy. Previous work has focused on identifying confounding factors, but a robust preoperative risk prediction model remains elusive. METHODS Using a dataset of 648 patients and 812 impacted mandibular third molars, we used least absolute shrinkage and selection operator (LASSO) to fit prediction models based on risk factors assessed at both the tooth and patient levels. In addition, we fitted multivariable logistic regression models with the Firth correction for generalized estimating equations (GEE). RESULTS The LASSO model for post-traumatic neuropathy identified distoangular impaction of ��� 45�� (odds ratio [OR]��=��2.9), proximity to the inferior alveolar nerve of ��� 3��mm (OR��=��1.9), disadvantageous curving (OR��=��1.4), and psychiatric conditions (OR��=��2.1) as predictors [area under the receiving operator characteristic curve (AUC)��=��0.75]. Among other complications analyzed, the LASSO model for bleeding identified deep embedding or full impaction (OR��=��1.8), psychiatric conditions (OR��=��1.3), and age (OR��=��0.9) as predictors (AUC��=��0.64). These associations between predictors and postoperative complications were fundamentally reinforced by the corresponding GEE models. CONCLUSIONS Our findings point to the predictability of post-traumatic neuropathy and bleeding based on tooth anatomy and patient characteristics, overall suggesting that preoperatively identifiable factors can predict the risk of adverse outcomes in the extraction of impacted mandibular third molars. CLINICAL SIGNIFICANCE Mandibular third molar extraction is both a routine procedure and a leading cause of trigeminal neuropathy. Prevention of post-traumatic neuropathy, aided by individualized preoperative risk prediction, is of high clinical relevance.

PURPOSE To investigate the use of platelet-rich fibrin for alveolar ridge preservation compared to natural healing, bone graft material and platelet-rich fibrin in combination with bone graft material. MATERIALS AND METHODS The present systematic review was conducted and reported according to the Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines. The review examined randomised controlled trials comparing the clinical outcomes of platelet-rich fibrin with those of other modalities for alveolar ridge preservation. Studies of third molar extraction site healing were excluded. The studies were classified into three categories: natural wound healing vs platelet-rich fibrin; bone graft material vs platelet-rich fibrin; and bone graft material vs bone graft material and platelet-rich fibrin. RESULTS From 179 articles identified, 16 randomised controlled trials were included. Owing to the heterogeneity of the investigated parameters, it was not possible to perform a meta-analysis. In total, 10 randomised controlled trials compared platelet-rich fibrin to natural wound healing, with seven of these demonstrating favourable outcomes to either limit postextraction dimensional changes or improve new bone formation in the platelet-rich fibrin group. Three of four studies comparing healing with bone graft material to platelet-rich fibrin found that the latter led to significantly greater horizontal or vertical bone resorption, and the bone graft material was more able to maintain the ridge dimensions. Two out of three randomised controlled trials investigating healing with both bone graft material and platelet-rich fibrin reported better outcomes using this combined approach than with bone graft material alone. All studies investigating soft tissue healing with platelet-rich fibrin demonstrated better outcomes in the platelet-rich fibrin group. CONCLUSIONS The majority of studies comparing healing with platelet-rich fibrin to natural healing concluded that the former more successfully limits postextraction dimensional changes than the latter. However, 75% of studies investigating platelet-rich fibrin vs bone graft material reported better results in the bone graft group with respect to its ability to maintain postextraction dimensional changes. The addition of platelet-rich fibrin to bone graft material may improve clinical outcomes, although data are limited.

Particulate autologous tooth roots are increasingly used for alveolar bone augmentation; however, the proteomic profile of acid dentin lysate and the respective cellular response have not been investigated. Here we show that TGF-β1 is among the 226 proteins of acid dentin lysate (ADL) prepared from porcine teeth. RNA sequencing identified 231 strongly regulated genes when gingival fibroblasts were exposed to ADL. Out of these genes, about one third required activation of the TGF-β receptor type I kinase including interleukin 11 (IL11) and NADPH oxidase 4 (NOX4). Reverse transcription-quantitative polymerase chain reaction and immunoassay confirmed the TGF-β-dependent expression of IL11 and NOX4. The activation of canonical TGF-β signaling by ADL was further confirmed by the phosphorylation of Smad3 and translocation of Smad2/3, using Western blot and immunofluorescence staining, respectively. Finally, we showed that TGF-β activity released from dentin by acid lysis adsorbs to titanium and collagen membranes. These findings suggest that dentin particles are a rich source of TGF-β causing a major response of gingival fibroblasts.

Objectives This study aims to compare the treatment outcomes of periodontal intrabony defects by using platelet-rich fibrin (PRF) with other commonly utilized modalities. Materials and methods The eligibility criteria comprised randomized controlled trials (RCTs) comparing the clinical outcomes of PRF with that of other modalities. Studies were classified into 10 categories as follows: (1) open flap debridement (OFD) alone versus OFD/PRF; (2) OFD/bone graft (OFD/BG) versus OFD/PRF; (3) OFD/BG versus OFD/BG/PRF; (4–6) OFD/barrier membrane (BM), OFD/PRP, or OFD/enamel matrix derivative (EMD) versus OFD/PRF; (7) OFD/EMD versus OFD/EMD/PRF; (8–10) OFD/PRF versus OFD/PRF/metformin, OFD/PRF/bisphosphonates, or OFD/PRF/statins. Weighted means and forest plots were calculated for probing depth (PD), clinical attachment level (CAL), and radiographic bone fill (RBF). Results From 551 articles identified, 27 RCTs were included. The use of OFD/PRF statistically significantly reduced PD and improved CAL and RBF when compared to OFD. No clinically significant differences were reported when OFD/BG was compared to OFD/PRF. The addition of PRF to OFD/BG led to significant improvements in CAL and RBF. No differences were reported between any of the following groups (OFD/BM, OFD/PRP, and OFD/EMD) when compared to OFD/PRF. No improvements were also reported when PRF was added to OFD/EMD. The addition of all three of the following biomolecules (metformin, bisphosphonates, and statins) to OFD/PRF led to statistically significant improvements of PD, CAL, and RBF. Conclusions The use of PRF significantly improved clinical outcomes in intrabony defects when compared to OFD alone with similar levels being observed between OFD/BG and OFD/PRF. Future research geared toward better understanding potential ways to enhance the regenerative properties of PRF with various small biomolecules may prove valuable for future clinical applications. Future research investigating PRF at histological level is also needed. Clinical relevance The use of PRF in conjunction with OFD statistically significantly improved PD, CAL, and RBF values, yielding to comparable outcomes to OFD/BG. The combination of PRF with bone grafts or small biomolecules may offer certain clinical advantages, thus warranting further investigations.

Allografts consisting of demineralized bone matrix (DBM) are supposed to retain the growth factors of native bone. However, it is not clear if transforming growth factor &beta, 1 (TGF-&beta, 1) is maintained in the acid-extracted human bone. To this aim, the aqueous solutions of supernatants and acid lysates of OraGRAFT&reg, Demineralized Cortical Particulate and OraGRAFT&reg, Prime were prepared. Exposing fibroblasts to the aqueous solution caused a TGF-&beta, receptor type I kinase-inhibitor SB431542-dependent increase in interleukin 11 (IL11), NADPH oxidase 4 (NOX4), and proteoglycan 4 (PRG4) expression. Interleukin 11 expression and the presence of TGF-&beta, 1 in the aqueous solutions were confirmed by immunoassay. Immunofluorescence further confirmed the nuclear translocation of Smad2/3 when fibroblasts were exposed to the aqueous solutions of both allografts. Moreover, allografts released matrix metalloprotease-2 activity and blocking proteases diminished the cellular TGF-&beta, response to the supernatant. These results suggest that TGF-&beta, is preserved upon the processing of OraGRAFT&reg, and released by proteolytic activity into the aqueous solution.

Purpose The purpose of this prospective clinical study was to evaluate the efficiency of alveolar ridge reconstruction with the lateral border of scapula (LBS) prior to implant placement and to assess onlay graft retention and bone resorption during a short term of function. Materials and methods A total of 25 partially or fully edentulous patients with severe alveolar bone atrophy received ridge reconstruction with grafts harvested from the LBS. Histologic analysis of bone grafts was performed. Six months after augmentation, patients underwent CBCT and received dental implants. After another 3 months, healing abutments and implant-supported dentures were placed. Patients were followed for an average of 24 months. Results Thirteen patients received primary bone grafting from LBS. Twelve patients experienced unsuccessful ridge reconstruction with other grafts before and were secondarily augmented with LBS. The average dimensions of LBS grafts were 6.3 × 2.3 × 1.2 cm. Histologic analysis confirmed the cortical nature of the graft. No donor-site complications occurred, and arm movements were restored within 2 weeks. Following augmentation, two patients had sutures disrupted that healed uneventfully after revision. The average resorption of LBS grafts after 6 months was 12.2% ± 3.0%. At the time of implant placement, the dimension of the ridge was 12.3 ± 2.0 mm and 6.9 ± 1.6 mm in height and width, respectively. The survival rate of the 174 implants placed was 98.3%. Conclusion LBS can be used as an alternative extraoral grafting site for extensive ridge reconstruction prior to implant placement.

Macrophage activation and osteoclastogenesis are hallmarks of inflammatory osteolysis and may be targeted by the local application of liquid platelet-rich fibrin (PRF). Liquid PRF is produced by a hard spin of blood in the absence of clot activators and anticoagulants, thereby generating an upper platelet-poor plasma (PPP) layer, a cell-rich buffy coat layer (BC; termed concentrated-PRF or C-PRF), and the remaining red clot (RC) layer. Heating PPP has been shown to generate an albumin gel (Alb-gel) that when mixed back with C-PRF generates Alb-PRF having extended working properties when implanted in vivo. Evidence has demonstrated that traditional solid PRF holds a potent anti-inflammatory capacity and reduces osteoclastogenesis. Whether liquid PRF is capable of also suppressing an inflammatory response and the formation of osteoclasts remains open. In the present study, RAW 264.7 and primary macrophages were exposed to lipopolysaccharides (LPS), lactoferrin, and agonists of Toll-like receptors (TLR3 and TLR7) in the presence or absence of lysates prepared by freeze-thawing of liquid PPP, BC, Alb-gel, and RC. For osteoclastogenesis, primary macrophages were exposed to receptor activator of nuclear factor kappa B ligand (RANKL), macrophage colony-stimulating factor (M-CSF), and human transforming growth factor-β1 (TGF-β1) in the presence or absence of PPP, BC, Alb-gel, RC lysates and hemoglobin. We show here that it is mainly the lysates prepared from PPP and BC that consistently reduced the agonist-induced expression of interleukin 6 (IL6) and cyclooxygenase-2 (COX2) in macrophages, as determined by RT-PCR and immunoassay. With respect to osteoclastogenesis, lysates from PPP and BC but also from RC, similar to hemoglobin, reduced the expression of osteoclast marker genes tartrate-resistant acid phosphatase (TRAP) and cathepsin K, as well as TRAP histochemical staining. These findings suggest that liquid PRF holds a potent in vitro heat-sensitive anti-inflammatory activity in macrophages that goes along with an inhibition of osteoclastogenesis.

PURPOSE Compaction of particulated grafts is done manually; thus, the effect of compression force on bone regeneration remains unclear. The aim of this study was to evaluate the impact of two different compression forces on the consolidation of particulated bovine hydroxyapatite. MATERIALS AND METHODS Two titanium cylinders were fixed on the calvarium of eight New Zealand rabbits. Both defects were filled with particulated bovine hydroxyapatite subjected to a compression force of 0.7 kg/cm2 or 1.6 kg/cm2 before being covered with a resorbable collagen membrane. A handheld device that uses a spring to control the compression force applied by the plugger was used. At 6 weeks, histomorphometry of the area immediately adjacent to the calvaria bone and to the collagen membrane was performed. RESULTS It was shown that next to the calvaria, the bone volume per tissue volume (BV/TV) was 29.0% ± 8.8% and 27.6% ± 8.2% at low and high compression force, respectively; the bone-to-biomaterial contact (BBC) was 58.2% ± 25.0% and 69.3% ± 22.9%, respectively (P > .05). In the corresponding area next to the collagen membrane, BV/TV was 4.9% ± 5.1% and 5.7% ± 4.7%, and the BBC was 18.3% ± 20.8% and 20.1% ± 15.9%, respectively (P > .05). In addition, the number and area of blood vessels were not significantly affected by compression force. CONCLUSION Both compression forces applied resulted in similar consolidation of bovine hydroxyapatite expressed by new bone formation and vascularization based on a rabbit calvaria augmentation model.

Collagen membranes and matrices being widely used in guided bone regeneration and soft tissue augmentation have characteristic properties based on their composition. The respective proteomic signatures have not been identified. Here, we performed a high-resolution shotgun proteomic analysis on two porcine collagen-based biomaterials designed for guided bone regeneration and soft tissue augmentation. Three lots each of a porcine-derived collagen membrane and a matrix derived from peritoneum and/or skin were digested and separated by nano-reverse-phase high-performance liquid chromatography. The peptides were subjected to mass spectrometric detection and analysis. A total of 37 proteins identified by two peptides were present in all collagen membranes and matrices, with 11 and 16 proteins being exclusively present in the membrane and matrix, respectively. The common extracellular matrix proteins include fibrillar collagens (COL1A1, COL1A2, COL2A1, COL3A1, COL5A1, COL5A2, COL5A3, COL11A2), non-fibrillar collagens (COL4A2, COL6A1, COL6A2, COL6A3, COL7A1, COL16A1, COL22A1), and leucine-rich repeat proteoglycans (DCN, LUM, BGN, PRELP, OGN). The structural proteins vimentin, actin-based microfilaments (ACTB), annexins (ANXA1, ANXA5), tubulins (TUBA1B, TUBB), and histones (H2A, H2B, H4) were also identified. Examples of membrane-only proteins are COL12A1 and COL14A1, and, of matrix only proteins, elastin (ELN). The proteomic signature thus revealed the similarities between but also some individual proteins of collagen membrane and matrix.

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.

MicroRNA‑21 (miR‑21) is a small non‑coding RNA that is differentially expressed during tooth development, particularly during amelogenesis. Although orthodontic tooth movement and the innate immune response are impaired, miR‑21 knockout mice demonstrate no obvious skeletal phenotype. However, the consequence of miR‑21 knockout on tooth phenotype and corresponding alveolar bone is unknown. The current study utilized landmark‑based geometric morphometrics to identify anatomical dissimilarities of the three lower and upper molars, and the corresponding alveolar bone, in miR‑21 knockout and wild‑type control mice. The anatomical structures were visualized by microcomputer tomography. A total of 36 and 38 landmarks were placed on mandibular and maxillary molars, respectively. For the alveolar bone, 16 landmarks were selected on both anatomical sites. General Procrustes analysis revealed significantly smaller molars and dimensions of the alveolar bone in the mandible of the miR‑21 knockout mice when compared with wild‑type controls (P=0.03 and P=0.04, respectively). The overall dimension of the mandible was reduced by the lack of miR‑21 (P=0.02). In the maxilla, the dimension of the alveolar bone was significant (P=0.02); however, this was not observed in the molars (P=0.36). Based on principal component analysis, no changes in shape for any of the anatomical sites were observed. Dental and skeletal jaw length were calculated and no prognathism was identified. However, the fluctuating asymmetry of the molars in the mandible and the maxilla was reduced in the miR‑21 knockout mice by 38 and 27%, respectively. Taken together, the results of the present study revealed that the molars in the mandible and the dimension of the respective alveolar bone were smaller in miR‑21 mice compared with wild‑type littermates, suggesting that miR‑21 influences tooth development.

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.

The preparation of platelet-rich fibrin (PRF) requires blood centrifugation to separate the yellow plasma from the red erythrocyte fraction. PRF membranes prepared from coagulated yellow plasma are then transferred to the defect sites to support tissue regeneration. During natural wound healing, however, it is the unfractionated blood clot (UBC) that fills the defect site. It is unclear whether centrifugation is necessary to prepare a blood-derived matrix that supports tissue regeneration. The aim of the present study was to compare lysates prepared from PRF and UBC based on bioassays and degradation of the respective membranes. We report here that lysates prepared from PRF and UBC membranes similarly activate TGF-β signaling, as indicated by the expression of interleukin 11 (IL-11), NADPH oxidase 4 (NOX-4) and proteoglycan 4 (PRG4) in gingival fibroblasts. Consistently, PRF and UBC lysates stimulated the phosphorylation and nuclear translocation of Smad3 in gingival fibroblasts. We further observed that PRF and UBC lysates have comparable anti-inflammatory activity, as shown by the reduction in lipopolysaccharide (LPS)-induced IL-6, inducible nitric oxidase synthase (iNOS) and cyclooxygenase 2 (COX-2) expression in RAW264.7 cells. Moreover, inflammation induced by Poly (1:C) HMW and FSL-1, which are agonists of Toll-like receptor (TLR) 3 and 2/6, respectively, was reduced by both PRF and UBC. PRF and UBC lysates reduced the nuclear translocation of p65 in LPS-induced RAW264.7 cells. In contrast to the similar activity observed in the bioassays, UBC membranes lack the structural integrity of PRF membranes, as indicated by the rapid and spontaneous disintegration of UBC membranes. We show here that the lysates prepared from PRF and UBC possess robust TGF-β and anti-inflammatory activity. However, visual inspection of the PRF and UBC membranes confirmed the negative impact of erythrocytes on the structural integrity of membranes prepared from whole blood. The data from the present study suggest that although both UBC and PRF have potent TGF-β and anti-inflammatory activity, UBC does not have the strength properties required to be used clinically to prepare applicable membranes. Thus, centrifugation is necessary to generate durable and clinically applicable blood-derived membranes.

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