Inhibition of osteosarcoma metastasis in vivo by targeted downregulation of MMP1 and MMP9.

Osteosarcoma (OS) mortality stems from lung metastases. Matrix metalloproteinases (MMPs) facilitate metastatic dissemination by degrading extracellular matrix components. Herein we studied the impact of targeted MMP downregulation on OS metastasis. Differential gene expression analysis of human OS cell lines revealed high MMP9 expression in the majority of OS cell lines. Furthermore, 143B, a metastatic OS cell line, exhibited increased MMP1 and MMP9 mRNA levels. Gene set enrichment analysis on metastatic and non-metastatic OS patient specimens indicated epithelial-mesenchymal transition as the most enriched gene set, with MMP9 displaying strong association to genes in this network. Using the same dataset, Kaplan-Meier analysis revealed a correlation between MMP1 expression and dismal patient survival. Hence, we undertook targeted suppression of MMP1 and MMP9 gene expression in OS cell lines. The ability of OS cells to migrate and form colonies was markedly reduced upon MMP1 and MMP9 downregulation, whereas their cell proliferation capacity remained intact. MMP9 downregulation decreased tumor growth and lung metastases area in an orthotopic mouse OS model. Consistently, human OS lung metastasis specimens displayed marked MMP9 protein expression. Our findings highlight the role of MMP1 and MMP9 in OS metastasis, warranting further exploration of simultaneous inhibition of MMPs for future OS therapeutics.

Tissue changes around dental implants installed in alveolar ridge preservation sites: A 1-year follow-up randomized controlled clinical trial.

OBJECTIVE: This study aimed to assess radiographic marginal bone changes 22 months post extraction, which is 1 year after implant loading in alveolar ridge preservation (ARP) sites grafted with a combination of collagen-embedded xenogenic bone substitute (DBBM-C) and collagen matrix (CMX), comparing them with implants placed in naturally healed sites. METHODS: This randomized controlled clinical trial was conducted over 22 months. Patients who needed a single tooth extraction and subsequent implant placement in nonmolar areas were enrolled. The test group received deproteinized bovine bone mineral with 10% collagen covered by a procaine collagen membrane, while the control group allowed spontaneous healing. Radiographic bone level changes were documented using periapical radiographs at implant placement and follow-up visits (6, 10, and 22 months postextraction). Early implant soft tissue exposure, clinical parameters, and patient-reported outcomes were recorded. RESULTS: Twenty-two out of 28 participants completed a 22-month follow-up, 9 in the test group and 13 in the control group. At 10-month postextraction follow-up, the mean MBL was 1.01 ± 1.04 mm in the treatment group and 0.81 ± 0.93 mm in the control group (p = 0.804). At 22 months, the mean MBL was 2.09 ± 1.03 mm in the treatment group and 1.58 ± 0.73 mm in the control group (p = 0.339). No statistically significant differences in probing depth (PD) and bleeding on probing (BOP) were found at the 22 -month follow-up as well. Soft tissue mean recession was observed in the control group (0.36 ± 0.84 mm), while no recession was found in the test group (p = 0.2). Early implant soft tissue exposure occurred in 33% of test group participants, while none was observed in the control group (p = 0.047). CONCLUSION: One year after implant loading, no significant differences in marginal bone resorption were found between implants placed in ARP-treated and naturally healed sites. However, ARP-treated sites exhibited early implant soft-tissue exposure, suggesting a possible impairment in soft tissue healing.

Oral fibroblasts rescue osteogenic differentiation of mesenchymal stem cells after exposure to Zoledronic acid in a paracrine effect.

Background: Medication-related osteonecrosis of the jaw is a serious complication that develops in oncologic patients treated with Zoledronic acid. Although used for over 30 years, the influence of Zoledronic acid on bone has been thoroughly investigated, mainly on osteoclasts. While decreasing osteoclast differentiation and function, for many years it was thought that Zoledronic acid increased osteoblast differentiation, thus increasing bone volume. Moreover, despite the influence of soft tissue on the bone healing process, the impact of zoledronic acid on the interaction between soft tissue and bone was not investigated. Aim: Our goal was to investigate the influence of Zoledronic Acid and soft tissue cells on osteogenic differentiation of mesenchymal stem cells (MSCs). Materials and methods: Osteogenic differentiation of MSCs was examined after exposure to Zoledronic Acid. To determine the influence of soft tissue cells on MSCs' osteogenic differentiation, conditioned media from keratinocytes and oral fibroblasts were added to osteogenic medium supplemented with Zoledronic Acid. Proteomic composition of keratinocytes' and fibroblasts' conditioned media were analyzed. Results: Zoledronic Acid decreased osteogenic differentiation of MSCs by seven-fold. The osteogenic differentiation of MSCs was restored by the supplementation of fibroblasts' conditioned medium to osteogenic medium, despite Zoledronic acid treatment. Five osteogenic proteins involved in the TGFß pathway were exclusively identified in fibroblasts' conditioned medium, suggesting their role in the rescue effect. Conclusion: Oral fibroblasts secrete proteins that enable osteogenic differentiation of MSCs in the presence of Zoledronic Acid.

Revealing leukocyte populations in human peri-implantitis and periodontitis using flow cytometry.

OBJECTIVE: To identify, quantify, and characterize leukocyte populations in PI and periodontitis using flow cytometry. METHODS: Fresh biopsies from human PI and periodontitis lesions were processed to a single-cell suspension. The immune cell types were identified using flow cytometry. RESULTS: Twenty-one biopsies were obtained and analyzed corresponding to fourteen PI and seven periodontitis samples. Participants' average age was 63.95 ± 14.77 years without a significant difference between PI and periodontitis patients, the female/male ratio was 8/12, and mean PD was 8.5 ± 2.17. High similarity was found between periodontitis and PI in the main immune cell types. Out of the leukocytes, the PMN proportion was 40% in PI and 33% in periodontitis. T-cells 22% in PI and 18% in periodontitis. Similar proportions of B-cells 10% and macrophages 6% were found in PI and periodontitis. Dendritic and NK cells were found in low proportions (~ 1%) in PI and periodontitis. T-cell sub-analysis showed that CD4-positive were more prevalent than CD8-positive in both diseases (CD4/CD8 ratio of 1.2). CONCLUSION: With the use of flow cytometry analysis, the leukocyte populations in human peri-implantitis and periodontitis were classified. In PI and periodontitis, we identified similar proportions of specific (CD4/CD8) and innate (dendritic and NK) immune cells. These results corroborate previous histological studies. CLINICAL RELEVANCE: Flow cytometry analysis can be used to identify and quantify immune cells in PI and periodontitis, including sub-classification of T cells (CD4/8) as well as detection of cells that require multiple markers for identification (such as dendritic cells).

Endothelial Progenitor Cells Promote Osteosarcoma Progression and Invasiveness via AKT/PI3K Signaling.

BACKGROUND: Osteosarcoma (OS) mortality is attributed to lung metastases. Endothelial progenitor cells (EPCs) mediate the angiogenic switch in several cancers. The spatial proximity between EPCs and OS in the bone led to the hypothesis that EPCs-osteosarcoma interactions may possibly promote OS progression and aggressiveness. METHODS: A PI3K inhibitor, Bevacizumab (an anti-VEGF-A antibody), and an anti-FGF2 antibody were added to the EPCs' conditioned medium (EPC-CM), and their impacts on OS cell (U2-OS and 143B) proliferation, migration, invasion, MMP9 expression, and AKT phosphorylation were determined. The autocrine role of VEGF-A was assessed using Bevacizumab treatment and VEGF-A silencing in OS cells. Toward this end, an orthotopic mouse OS model was established. Mouse and human tumors were immunolabeled with antibodies to the abovementioned factors. RESULTS: EPC-CM enhanced osteosarcoma MMP9 expression, invasiveness, and migration via the PI3K/AKT pathway. The addition of Bevacizumab and an anti-FGF2 antibody to the EPC-CM diminished OS cell migration. The autocrine role of VEGF-A was assessed using Bevacizumab and VEGF-A silencing in OS cells, resulting in decreased AKT phosphorylation and, consequently, diminished invasiveness and migration. Consistently, OS xenografts in mice displayed high VEGF-A and FGF2 levels. Remarkably, lung metastasis specimens derived from OS patients exhibited marked immunolabeling of CD31, VEGF-A, and FGF2. Conclusions: EPCs promote OS progression not only by physically incorporating into blood vessels, but also by secreting cytokines, which act via paracrine signaling. EPCs induced in vitro MMP9 overexpression, invasion, and migration. Additional animal studies are warranted to further expand these results. These findings may pave the way toward the development of novel EPCs-targeted therapeutics aimed at blocking OS metastasis.

Comparing Operators and Imaging Techniques When Performing Trans-Crestal Sinus Augmentation: A Pilot Study.

The aim of this article was to compare baseline residual ridge height using Cone-beam Computed Tomography (CBCT) and panoramic radiographs. A secondary aim was to examine the magnitude of vertical bone gain 6 months after trans-crestal sinus augmentation and compare it between operators. Thirty patients, who underwent trans-crestal sinus augmentation simultaneously with dental implant placement, were included in this retrospective analysis. Surgeries were done by 2 experienced surgeons (EM and EG) using the same surgical protocol and materials. Preoperative residual ridge height was measured on panoramic and CBCT images. The final bone height and the magnitude of the vertical augmentation were measured on panoramic X ray taken 6 months after surgery. Mean residual ridge height measured preoperatively using CBCT was 6.07 ± 1.38 mm, whereas these same measurements on the panoramic radiographs yielded similar results (6.08 ± 1.43 mm), which were statistically insignificant (P = .535). Postoperative healing was uneventful in all cases. All 30 implants were successfully osseointegrated at 6 months. The mean overall final bone height was 12.87 ± 1.39 mm (12.61 ± 1.21 and 13.39 ± 1.63 mm for operators EM and EG, respectively; P = .19). Likewise, mean postoperative bone height gain was 6.78 ± 1.57 mm, which was 6.68 ± 1.32 and 6.99 ± 2.06 mm for operators EM and EG, respectively (P = .66). A moderate positive correlation was found between residual bone height and final bone height (r = 0.43, P = .002). A moderate negative correlation was found between residual bone height and augmented bone height (r = -0.53, P = .002). Sinus augmentation performed trans-crestally produce consistent results with minimal interoperator differences between experienced clinicians. Both CBCT and panoramic radiographs produced similar assessment of the preoperative residual bone height.

Palatal soft tissue thickness around dental implants and natural teeth in health and disease: A cross sectional study.

BACKGROUND: Previous studies focused on the influence of buccal mucosa thickness on peri-implant bone loss and inflammation, with inconclusive results. We observed substantially thicker palatal mucosal tissues at peri-implantitis sites. Therefore, we hypothesize that thick palatal peri-implant mucosa may be associated with deeper pockets and disease severity. PURPOSE: To compare the thickness of the palatal tissue between natural teeth and implants in periodontal health and disease. METHODS: Adult, non-smoker, healthy patients who visited our department for periodontal examination or treatment with restored implants in the posterior maxilla were recruited. Probing depth (PD), plaque index (PI), gingival index (GI) and radiographic measurements were recorded around implant and the contralateral tooth. Palatal tissue thickness was measured using a 30G needle that was inserted perpendicular into the mucosa at the bottom of the periodontal/peri-implant pocket and 3 mm coronally. Differences in the palatal tissue thickness between teeth and implants (in the same patient) was performed using t-test; as well as between peri-implantitis and non-peri-implantitis sites (among patients). RESULTS: Sixty patients were included. Thirty-four implants were diagnosed with peri-implantitis and 26 healthy/mucositis implants with corresponding 24 healthy/gingivitis teeth and 36 teeth with attachment loss. Mean PD was higher around implants (4.47 ± 1.57 mm) than teeth (3.61 ± 1.23 mm, p = 0.001). The thickness of implants' palatal mucosa was higher than in teeth, at the base of the pocket and 3 mm coronally (4.58 ± 1.38 mm vs. 3.01 ± 1.11, p = 0.000; 3.58 ± 2.15 vs. 1.89 ± 1.11, p = 0.000, respectively). Mean palatal tissue thickness was 4.32 ± 2.35 mm for the peri-implantitis group while only 2.61 ± 1.39 in healthy implants, 3 mm coronal to the base of the pocket (p = 0.001). Palatal thickness at peri-implantitis sites was higher (4.32 ± 2.35) compared to periodontitis sites (2.23 ± 0.93), p = 0.000. Implant sites with palatal mucosa >4 mm (n = 32) had deeper mean pockets (5.58 ± 1.98) compared with thinner (≤4 mm) sites (n = 28) (4.48 ± 1.18, p = 0.018). CONCLUSION: Thicker palatal tissue around implants is associated with deeper palatal pockets. Thick palatal tissue was found around implants diagnosed with peri-implantitis.

IFN-ß mediates the anti-osteoclastic effect of bisphosphonates and dexamethasone.

Zoledronic acid (Zol) is a potent bisphosphonate that inhibits the differentiation of monocytes into osteoclasts. It is often used in combination with dexamethasone (Dex), a glucocorticoid that promotes the resolution of inflammation, to treat malignant diseases, such as multiple myeloma. This treatment can result in bone pathologies, namely medication related osteonecrosis of the jaw, with a poor understanding of the molecular mechanism on monocyte differentiation. IFN-ß is a pro-resolving cytokine well-known as an osteoclast differentiation inhibitor. Here, we explored whether Zol and/or Dex regulate macrophage osteoclastic differentiation via IFN-ß. RAW 264.7 and peritoneal macrophages were treated with Zol and/or Dex for 4-24 h, and IFN-ß secretion was examined by ELISA, while the IFN stimulated gene (ISG) 15 expression was evaluated by Western blotting. RANKL-induced osteoclastogenesis of RAW 264.7 cells was determined by TRAP staining following treatment with Zol+Dex or IFN-ß and anti-IFN-ß antibodies. We found only the combination of Zol and Dex increased IFN-ß secretion by RAW 264.7 macrophages at 4 h and, correspondingly, ISG15 expression in these cells at 24 h. Moreover, Zol+Dex blocked osteoclast differentiation to a similar extent as recombinant IFN-ß. Neutralizing anti-IFN-ß antibodies reversed the effect of Zol+Dex on ISG15 expression and partially recovered osteoclastic differentiation induced by each drug alone or in combination. Finally, we found Zol+Dex also induced IFN-ß expression in peritoneal resolution phase macrophages, suggesting these drugs might be used to enhance the resolution of acute inflammation. Altogether, our findings suggest Zol+Dex block the differentiation of osteoclasts through the expression of IFN-ß. Revealing the molecular pathway behind this regulation may lead to the development of IFN-ß-based therapy to inhibit osteoclastogenesis in multiple myeloma patients.

Digital photometric analysis of gingival response to periodontal treatment.

OBJECTIVES: The color is a major factor in determining inflammation status in most gingival indices. Current indices have limitations mainly due to subjective nature. Digital color analysis can provide objective and accurate measurements. Thus, the present study aimed to assess by digital tool the gingival color in the different stages of an active periodontal treatment. METHODS: Forty patients (19 males and 21 females) diagnosed with periodontitis (stage III/ IV, grade C) and treated surgically were included in the study. Clinical data (probing depth, bleeding on probing, clinical attachment level, gingival index, and gingival recession) and photographs by digital single-lens-reflex (DSLR) camera were recorded before initial periodontal treatment, which included scaling and root surface debridement (T0); the same parameters were then re-evaluated 6-8 weeks (T1) and 3 months after periodontal surgery (regenerative/resective) (T2). Differences between clinical parameters were calculated. The color space defined by the International Commission on Illumination (CIELab) was used to analyze gingival color. RESULTS: In 56 periodontal surgical sites, 168 photographs were taken. The a*-value of the CIELab color system (higher a*- value translate to a stronger red color) was significantly reduced between T0 to T1 and further decreased at T2 (32.01, 29.28, and 27.45 respectively). Significant improvement in clinical parameters were found between T0 to T1 and T1 to T2. Sub-analysis of two distinct surgical interventions revealed that only regenerative procedure improved the a*-value, which was significantly correlated with pocket depth reduction. CONCLUSIONS: Photometric analysis can be used to assess gingival color change during periodontal treatment of patients with periodontitis. CLINICAL SIGNIFICANCE: Gingival inflammation is a major factor in periodontal assessment; nevertheless, all current gingival inflammation indices are partially subjective and only semi-quantitative. The digital photometric analysis may allow for accurate and objective gingival color assessment during periodontal treatment.

Volumetric assessment of changes in the alveolar ridge dimension following guided bone regeneration using a combination freeze-dried bone allograft with collagen membrane or novel resorbable scaffold: A prospective two-center clinical trial.

BACKGROUND: The aim of this study was to examine osseous changes following lateral bone augmentation using a novel scaffold (OV) alone and compare it to combination therapy using freeze-dried bone allograft (FDBA) and resorbable collagen membrane (FDBA/CM). METHODS: Thirty patients completed this 9-months prospective two-center cohort clinical trial. Before surgery and 9-months re-entry, linear measurements were performed, and impressions taken. Cone-beam computed tomography (CBCT) were done at baseline and 9 months. DICOM slice data were converted into volumetric images using 3D Slicer. Following 3D volumetric image construction, pre- and post-op Standard Triangle Language files were superimposed and volumetric data were extracted for a 10-mm region of interest. Linear measurements were compared similarly. RESULTS: Baseline clinical parameters were similar in both groups (4.22 and 4.53 mm for OV and FDBA/CM at -2 mm, respectively). Following treatment, vertical distance from the stent had changed minimally (-0.36 and -0.12 mm, respectively). Similarly, lateral bone gain ranged from 0 to 0.4 mm, for both groups. To the contrary, the CBCT measurements showed a significantly greater increase in horizontal width in the control at -2 mm (0.95 ± 0.2 mm) compared with -0.62 mm for the OV (P = 0.000). Similar changes were observed at -5 mm (0.63 and -0.41 mm, respectively, P = 0.01). Sites volume had increased from 266 ± 149 mm3 to 360 ± 138 mm3 (P = 0.001) for FDBA/CM with negligible changes for OV (from 334 to 335 mm3 , P = 0.952). these between-group changes being statistically significant (P = 0.023). CONCLUSION: FDBA/CM yielded better albeit moderate increase in the volume of the edentulous ridge, while OV scaffolds failed to produce similar results.

Severe Complications Following Maxillary Sinus Augmentation Using Poly L-lactide-co-ε-caprolactone-Coated Bovine Bone: A Retrospective Study.

PURPOSE: To describe the postoperative complications following lateral wall sinus augmentation using (poly L-lactideco-ε-caprolactone; PLCL) and natural polysaccharides polymers-coated bovine bone (PBB). The secondary aims were to examine histologic findings and to propose complication management alternatives. MATERIALS AND METHODS: This retrospective study included 61 subjects who underwent 67 lateral wall sinus augmentation procedures using PBB in the standard protocol. In cases that presented complications, treatment included additional antibiotic therapy, implant removal, or sinus reentry and total removal of the grafting material. In three cases, biopsy specimens were taken from the sinuses, and histologic analyses were performed. RESULTS: The prevalence of postoperative complications was 32.8% (22 of 67 cases) in 18 of the patients (29.5%). The most prevalent symptoms were persistent pain (68.2%), swelling (63.6%), and oroantral fistula (54.5%). Radiographic signs appeared in 45.5% of the complications. A total of 24 implants failed; thus, an overall 80.3% survival rate was established at 19 months. The vast majority of complications (86.4%) were treated eventually with reentry surgery and revealed that the sinus was full with granulation tissue surrounding pieces of a nonossified rubber-like material. In cases where implants were placed, nonosseointegrated implants were surrounded by soft tissue. The sinus was cleaned thoroughly; the graft material remnants were removed together with inflamed parts of the sinus membrane, followed by chlorhexidine and saline lavages. In the biopsy specimens taken from the sinus cavity, there were no histologic features of new bone formation around the grafted material. CONCLUSION: Lateral wall maxillary sinus augmentation using PBB was associated with an acute sinus infection histologic appearance and with a 7-times-higher failure rate compared with previous reports. This serious adverse event suggests that PBB cannot be recommended for maxillary sinus augmentations.

The effect of a surgeon's position on the axial inclination of dental implants placed freehand: a single-blind study.

OBJECTIVES: To examine whether a surgeons' position affects the axial angulation of dental implants placed freehand. METHOD AND MATERIALS: Implants' axial angulation was assessed on digital panoramic radiographs. An occlusal line was plotted based on the neighboring teeth/crowns. The mesial angle between the long axis of the implant and the occlusal line was measured. In addition, post-hoc ideal implant-positioning planning was done on the panoramic digital image, and the angle of the intersection between the long axis of the actual and post-hoc ideal implant was measured. Student t test for unpaired observations and the Kolmogorov-Smirnov nonparametric tests were utilized to compare the ipsi- and contralateral sides and between clinicians. RESULTS: Seventy-seven patients (149 implants) were eligible for the study. Implants had slight mesial inclination (mean 97.7 ± 8.7 degrees) which was similar for both the ipsi- (98.2 ± 8.4 degrees) and contralateral sides (97.2 ± 9.1 degrees), P = .491. For the post-hoc planning versus actual placement comparison, the overall median (interquartile range) of implant angular deviation was minimal (-0.25 degrees [-2.98, +3.47]). This was true for both the ipsilateral (-0.5 degrees [-2.9, +2.9]) and contralateral (-0.2 [-4.2, +5.4]) sides, P = .55. For the actual versus post-hoc planning, most observations clustered around the midline (zero to minimal deviation), while for the implant to occlusal plane angle, a tendency towards slight mesial angulation was observed. CONCLUSIONS: Dental implants placed freehand by experienced clinicians have only slight axial deviation as measured from post-hoc optimal position. Implants placed in ipsilateral and contralateral sides and by left- and right-dominant-hand clinicians had similar angulations.

Endothelial progenitors increase vascularization and improve fibroblasts function that prevent medication-related osteonecrosis of the jaw.

OBJECTIVES: In a previous rat model, MRONJ occurrence was 50%. Our aim was to investigate the potential of endothelial progenitor cells (EPCs) to improve fibroblasts function and prevent MRONJ. METHODS: Human gingival fibroblasts were cultured with EPC-conditioned media (EPC-CM); endothelial growth media (EGM-2) or DMEM followed by incubation with 10 µM zoledronic (ZOL) and dexamethasone (DEX). Cell proliferation and migration were assessed by XTT and scratch wound healing assays. In vivo, ten Lewis rats were treated weekly with ZOL and DEX for 11 weeks. After a week, EPCs or EGM-2 were injected to the gingiva around the molars. At 3 weeks, bilateral molars were extracted. After 8 weeks, wound healing was assessed, and serum VEGF and blood vessels were quantified. RESULTS: ZOL/DEX significantly reduced fibroblasts proliferation and wound healing. Treatment with EPC-CM before ZOL/DEX improved cell proliferation, and scratch healing (p = .007, p = .023). In vivo, local EPC injection before tooth extraction increased serum VEGF (p = .01) and soft tissue vascularization (p = .05). Normal healing was similar (80%) in EPCs and EGM-2 groups. CONCLUSION: EPC rescued fibroblasts from the cytotoxic effect of ZOL/DEX and elevated serum VEGF and vessel density that might reduce MRONJ occurrence to 20% compared to 50% in a similar model.

The Paracrine Role of Endothelial Cells in Bone Formation via CXCR4/SDF-1 Pathway.

Vascularization is a prerequisite for bone formation. Endothelial progenitor cells (EPCs) stimulate bone formation by creating a vascular network. Moreover, EPCs secrete various bioactive molecules that may regulate bone formation. The aim of this research was to shed light on the pathways of EPCs in bone formation. In a subcutaneous nude mouse ectopic bone model, the transplantation of human EPCs onto ß-TCP scaffold increased angiogenesis (p < 0.001) and mineralization (p < 0.01), compared to human neonatal dermal fibroblasts (HNDF group) and a-cellular scaffold transplantation (ß-TCP group). Human EPCs were lining blood vessels lumen; however, the majority of the vessels originated from endogenous mouse endothelial cells at a higher level in the EPC group (p < 01). Ectopic mineralization was mostly found in the EPCs group, and can be attributed to the recruitment of endogenous mesenchymal cells ten days after transplantation (p < 0.0001). Stromal derived factor-1 gene was expressed at high levels in EPCs and controlled the migration of mesenchymal and endothelial cells towards EPC conditioned medium in vitro. Blocking SDF-1 receptors on both cells abolished cell migration. In conclusion, EPCs contribute to osteogenesis mainly by the secretion of SDF-1, that stimulates homing of endothelial and mesenchymal cells. This data may be used to accelerate bone formation in the future.

Emergency department visits at Rambam health care campus, Israel: non-trauma related dental conditions.

OBJECTIVES: Studies of emergency department (ED) visits for non-traumatic dental conditions (NTDCs) have been carried out in the USA and Canada. In Israel, there is a shortage of such studies. In the current retrospective study, we report on the frequency and distribution of NTDCs ED visits at Rambam Health Care Campus (Rambam), in Haifa, which is an academic hospital serving more than 2.4 million residents of Northern Israel. MATERIALS AND METHODS: The data concerning ED visits at Rambam between 2010 and 2017 were obtained retrospectively from Rambam's computerized clinical and personal database of adult patients (≥18 years) visiting the ED for NTDCs. RESULTS: Overall, 1.8% of the patients who visited the Rambam ED, were identified as presenting with NTDCs. From 2010 until 2017, the number of NTDCs admissions increased by 45%, while the total ED admissions rose by 16%. The average waiting time for maxillofacial consultations for patients with NTDCs increased from 102 min in 2010 to 138 min in 2017. The busiest hours in the ED for NTDCs were during the morning shifts (47% of daily visits). CONCLUSIONS: The results of the study show that systemic and conceptual changes are needed to reduce the number of non-trauma related applications to ED.These changes can be by increasing the number of personnel or by introducing recent advances such as tele-medicine for prescreening of patients. This change calls for a greater involvement of the health policy leaders to provide alternative solutions for emergency dental care.

Socket Preservation Using Xenograft Does Not Impair Implant Primary Stability in Sheep: Clinical, Histological, and Histomorphometric Study.

Implant primary stability, which depends mainly on the amount and quality of bone, is important for implant survival. Socket preservation aims to reduce bone volumetric changes after tooth extraction. This animal study aims to examine whether preserving a ridge by using xenograft impairs the primary stability of the implant. Eighteen artificial bone defects were prepared in 4 sheep (5- and 8-mm length). Defects were randomly grafted with xenografts: Bio-Oss (BO), Bio-Active bone (BB), or left for natural healing (control). After 8 weeks, bone biopsy was harvested and dental implants installed. During installation, peak insertion torque (IT) was measured by hand ratchet, and primary stability by the Osstell method. Histomorphometric analysis showed a higher percentage of new bone formation in the naturally healed defects compared to sites with xenograft (control: 68.66 ± 4.5%, BB: 48.75 ± 4.34%, BO: 50.33 ± 4.0%). Connective tissue portion was higher in the BO and BB groups compared to control (44.25 ± 2.98%, 41 ± 6%, and 31.33 ± 4.5%, P < .05, respectively). Residual grafting material was similar in BO and BB (7 ± 2.44%, 8.66 ± 2.1%, respectively). Mean IT and implant stability quotient (ISQ) values were not statistically different among the groups. A positive correlation was found between IT and ISQ (r = 0.65, P = 0). In conclusion, previously grafted defects with xenograft did not influence primary stability and implant insertion torque in delayed implant placement. These results may be attributed to a relatively high bone fill of the defect (∼50%) 2 months after grafting.

Endothelial Progenitor Cells inhibit jaw osteonecrosis in a rat model: A major adverse effect of bisphosphonate therapy.

Medication-related osteonecrosis of the jaw (MRONJ) is a serious adverse effect of antiresorptive and antiangiogenic therapies. MRONJ is identified by chronic wounds in the oral mucosa associated with exposed necrotic bone. We hypothesized that zoledronic acid (ZOL) impairs keratinocyte and fibroblast function and reduces soft tissue vascularization; therefore, treating MRONJ with proangiogenic cells may benefit MRONJ patients. The effect of ZOL and dexamethasone (DEX) on gingival fibroblasts and keratinocytes was investigated. In-vitro, ZOL inhibited fibroblast and keratinocyte proliferation, delaying scratch healing. In-vivo, exposed bone was detected at tooth extraction sites, mainly in ZOL(+)/DEX(+) rats; and was associated with significantly decreased soft tissue vascularization, serum-VEGF, and tissue-VEGF. Local injection of early and late endothelial progenitor cells (EPCs) healed 13 of 14 MRONJ lesions compared with 2/7 lesions in the mesenchymal stem cells, and 2/6, in culture-medium group. The EPCs reduced necrotic bone area, increased serum and tissue VEGF levels. EPCs engraftment was minimal, suggesting their paracrine role in MRONJ healing. The EPC-conditioned medium improved scratch healing of keratinocytes and fibroblasts via VEGF pathway and elevated mRNA of VEGFA and collagen1A1. In conclusion, a novel MRONJ treatment with EPCs, increased vascularization and improved epithelial and fibroblast functions as well as cured the lesion.

Predicting Angiogenesis by Endothelial Progenitor Cells Relying on In-Vitro Function Assays and VEGFR-2 Expression Levels.

Clinical trials have demonstrated the safety and efficacy of autologous endothelial progenitor cell (EPC) therapy in various diseases. Since EPCs' functions are influenced by genetic, systemic and environmental factors, the therapeutic potential of each individual EPCs is unknown and may affect treatment outcome. Therefore, our aim was to compare EPCs function among healthy donors in order to predict blood vessel formation (angiogenesis) before autologous EPC transplantation. Human EPCs were isolated from the blood of ten volunteers. EPCs proliferation rate, chemoattractant ability, and CXCR4 mRNA levels were different among donors (p < 0.0001, p < 0.01, p < 0.001, respectively). A positive correlation was found between SDF-1, CXCR4, and EPCs proliferation (R = 0.736, p < 0.05 and R = 0.8, p < 0.01, respectively). In-vivo, blood vessels were counted ten days after EPCs transplantation in a subcutaneous mouse model. Mean vessel density was different among donors (p = 0.0001); nevertheless, donors with the lowest vessel densities were higher compared to control (p < 0.05). Finally, using a linear regression model, a mathematical equation was generated to predict blood vessel density relying on: (i) EPCs chemoattractivity, and (ii) VEGFR-2 mRNA levels. Results reveal differences in EPCs functions among healthy individuals, emphasizing the need for a potency assay to pave the way for standardized research and clinical use of human EPCs.

Use of transforming growth factor-ß loaded onto ß-tricalcium phosphate scaffold in a bone regeneration rat calvaria model.

BACKGROUND: Transforming growth factor-ß (TGF-ß1 ) enhances mesenchymal stem cell (MSC) differentiation into osteoblasts. PURPOSE: The aim of the study was to assess whether TGF-ß1 loaded onto ß-tricalcium phosphate (ß-TCP) synthetic scaffold enhances bone regeneration in a rat calvaria model. The release kinetics of TGF-ß1 from ß-TCP scaffold was evaluated in vitro. MATERIALS AND METHODS: TGF-ß1 in various concentrations (1-40 ng/mL) was loaded onto the ß-TCP scaffold, and release kinetics was monitored by ELISA. The effect of TGF-ß1 on the proliferation of MSCs was assessed using AlamarBlue, and MSC differentiation was evaluated by Alizarin Red quantification assay.Bone augmentation following transplantation of TGF-ß1 loaded onto ß-TCP in a rat calvaria model was evaluated in vivo. RESULTS: Greater TGF-ß1 release from the 40 ng/mL concentration was found. A suppressive effect of TGF-ß on the MSCs proliferation was observed with maximum inhibition obtained with 40 ng/mL compared to the control group (P = .028). A positive effect on MSCs osteogenic differentiation was found.Bone height and bone area fraction in vivo were similar with or without TGF-ß1 ; however, blood vessel density and degradation of the scaffold were significantly higher in the TGF-ß1 group. CONCLUSION: TGF-ß1 adsorbed to ß-TCP stimulated angiogenesis and scaffold degradation that may enhance bone formation.

Prospective randomized controlled clinical trial to compare hard tissue changes following socket preservation using alloplasts, xenografts vs no grafting: Clinical and histological findings.

PURPOSE: To compare dimensional changes and bone quality of two different grafting materials used for socket preservation. MATERIALS AND METHODS: Thirty-three patients requiring extraction were recruited and randomly assigned to receive: biphasic calcium sulfate/ hydroxyapatite (BCS/HA); bovine derived xenograft (BDX) or no grafting (Control). Ridge width (at -3 and -6 mm) and vertical distance from a stent were measured at the time of extraction/grafting. Measurements were repeated at reentry and core biopsies were harvested. RESULTS: Baseline vertical distance for the BDX, C and BCS/HA groups were 7.45 ± 3.1, 7.69 ± 4.2, and 6.75 ± 3.5 mm, respectively (P = .830). Post-op, C group had greater vertical loss (1.71 ± 0.4 mm) compared to BCS/HA (0.65 ± 0.5) and BDX (0.25 ± 0.2 mm), P = .059. Mean baseline width at -3 mm was 8.69 ± 1.1 mm, 8.31 ± 1.4 mm, and 9.0 ± 1.1 mm, respectively (P = .509). Post-op, this width was reduced by 2.96 ± 0.3 mm (C), 1.56 ± 0.4 mm (BDX), and 0.5 ± 0.4 mm (BCS/HA), P = .001. Mean ridge width at -6 mm for the C (6.5 ± 1.7 mm) was significantly smaller than BCS/HA (7.95 ± 2.8 mm) and BDX (8.85 ± 1.9 mm), P = .043. Histologically, the BDX group had greater residual scaffold material and less vital bone compared to the BCS/HA group. Pain scores were relatively low for all groups. CONCLUSIONS: BCS/HA may be used for socket preservation with similar or better results compared to BDX. The significance of greater residual scaffold found in the BDX group is yet to be determined.