Periodontal regenerative therapy with enamel matrix derivative in the treatment of intrabony defects: a prospective 2-year study

Abstract Objective To date, enamel matrix derivative (EMD) has been considered to be one of the few biomaterials for clinical use capable of demonstrating true periodontal regeneration. The aim of this two-center prospective clinical study was to evaluate 2-year outcome of periodontal regenerative therapy using EMD in the treatment of intrabony defects, performed as an ‘advanced medical treatment’ under the national healthcare system in Japan. Results Patients with chronic periodontitis who have completed initial periodontal therapy at either of the two dental school clinics were enrolled. Each contributed at least one intrabony defect of ≥3 mm in depth. During surgery, EMD was applied to the defect following debridement. Twenty-two participants (mean age 55.2 years old, 9 men and 13 women) completed 2-year reevaluation, and a total of 42 defects were subjected to data analysis. Mean gains in clinical attachment level (CAL) at 1 and 2 years were 2.9 mm (38% of baseline CAL) and 3.1 mm (41%), respectively, both showing a significant improvement from baseline. There was also a significant reduction in probing depth (PD): mean reductions at 1 and 2 years were 3.2 and 3.3 mm, respectively. There was a progressive improvement in the mean percentages of bone fill from 26% at 1 year to 36% at 2 years. No significant difference in CAL gain at 2 years was found between 3-wall bone defects and other defect types combined. In multiple regression analysis, the baseline PD was significantly associated with CAL gain at 2 years. In this population of patients, the treatment of intrabony defects with EMD yielded clinically favorable outcomes, as assessed by periodontal and radiographical parameters, over a period of 2 years

MicroRNA-214 Promotes Apoptosis in Canine Hemangiosarcoma by Targeting the COP1-p53 Axis

<div><p>MicroRNA-214 regulates both angiogenic function in endothelial cells and apoptosis in various cancers. However, the regulation and function of miR-214 is unclear in canine hemangiosarcoma, which is a spontaneous model of human angiosarcoma. The expression and functional roles of miR-214 in canine hemangiosarcoma were presently explored by performing miRNA TaqMan qRT-PCR and transfecting cells with synthetic microRNA. Here, we report that miR-214 was significantly down-regulated in the cell lines used and in clinical samples of canine hemangiosarcoma. Restoration of miR-214 expression reduced cell growth and induced apoptosis in canine hemangiosarcoma cell lines through transcriptional activation of p53-regulated genes although miR-214 had a slight effect of growth inhibition on normal endothelial cells. We identified COP1, which is a critical negative regulator of p53, as a novel direct target of miR-214. COP1 was overexpressed and the specific COP1 knockdown induced apoptosis through transcriptional activation of p53-regulated genes as well as did miR-214-transfection in HSA cell lines. Furthermore, p53 knockdown abolished the miR-214-COP1-mediated apoptosis; thus, miR-214 and COP1 regulated apoptosis through controlling p53 in HSA. In conclusion, miR-214 functioned as a tumor suppressor in canine hemangiosarcoma by inducing apoptosis through recovering the function of p53. miR-214 down-regulation and COP1 overexpression is likely to contribute to tumorigenesis of HSA. Therefore, targeting miR-214-COP1-p53 axis would possibly be a novel effective strategy for treatment of canine hemangiosarcoma and capable of being applied to the development of novel therapeutics for human angiosarcoma.</p></div

Periodontal Regenerative Therapy Using rhFGF-2 and Deproteinized Bovine Bone Mineral versus rhFGF-2 Alone: 4-Year Extended Follow-Up of a Randomized Controlled Trial

The aim of this study was to evaluate longitudinal outcomes of recombinant human fibroblast growth factor (rhFGF)-2 plus deproteinized bovine bone mineral (DBBM) therapy in comparison with rhFGF-2 alone for treating periodontal intrabony defects. This study describes 4-year follow-up outcomes of the original randomized controlled trial. Intrabony defects in periodontitis patients were treated with rhFGF-2 (control) or rhFGF-2 plus DBBM (test). Clinical, radiographic, and patient-reported outcome (PRO) measures were used to evaluate the outcomes. Thirty-two sites were able to be followed up. At 4 years postoperatively, clinical attachment level (CAL) gains in the test and control groups were 3.5 &plusmn; 1.4 mm and 2.7 &plusmn; 1.4 mm, respectively, showing significant improvement from preoperative values but no difference between groups. Both groups showed an increase in radiographic bone fill (RBF) over time. At 4 years, the mean value for RBF in the test group (62%) was significantly greater than that in the control group (42%). In 1&ndash;2-wall defects, the test treatment yielded significantly greater RBF than the control treatment. No significant difference in PRO scores was noted between the groups. Although no significant difference in CAL gain was found between the groups at the 4-year follow-up, the combination treatment significantly enhanced RBF. Favorable clinical, radiographic outcomes, and PRO in both groups can be maintained for at least 4 years

miR-214 decreased the number of viable cells and induced apoptosis in HSA cell lines.

<p>(A) Cell viability was assessed by performing the MTT assay. miR-214-transfection decreased the number of viable cells in HSA cell lines and that of control EC. However, the degree of growth inhibition was slight for the control EC compared with that for the HSA cell lines. All data are presented as the mean of triplicate experiments with error bars indicating the s.d. (Two-tailed t-test; *p<0.05, **p<0.01). (B) Morphological assessment of nuclei by Hoechst 33342 nuclear staining. miR-214 increased the number of cells with fragmented nuclei (white arrow heads) dose-dependently in HSA cell lines, whereas it caused no morphological changes in the nuclei of the control EC. Scale bars in the photographs indicate 25 μm. The graph shows the percentage of cells with fragmented nuclei among a total 500 cells. All data are presented as the mean of triplicate experiments with error bars indicating the s.d. (Two-tailed t-test; *p<0.05, **p<0.01). (C) Apoptotic cell count by Annexin V/ PI double staining. Annexin V-positive/ PI-negative and Annexin V-positive/ PI-positive cells represent early and late-phase apoptosis, respectively. miR-214 increased both early and late-phase apoptosis in HSA cell lines. However, miR-214 hardly induced apoptosis in the control EC. All data are presented as the mean of triplicate experiments with error bars indicating the s.d. The statistical significances stated were referred to the entire Annexin/PI population (Two-tailed t-test; *p<0.05, **p<0.01). (D) Immunoblotting for caspase-3 active form and PARP proform. miR-214 increased the amount of the active form of caspase-3 and cleavage of PARP proform dose-dependently in HSA cell lines (JuB2, Re12 and Ud6) although significant changes were not observed in control EC (CnAOEC) transfected by miR-214. β-actin was used for normalization of the amount of sample loaded. (E) Cell cycle analysis. miR-214 increased the sub-G fraction and decreased S and G₂/M fractions in all HSA cell lines, indicating that miR-214 induced apoptosis in HSA cell lines; however, only slight decrease of S fraction were observed in control EC. The histograms show the representative data of each cell line transfected by 10 nM of miR-214 or control RNA in the triplicated analysis. The data of bar graphs are presented as the mean of triplicate experiments with error bars indicating the s.d. (Two-tailed t-test; *p<0.05, **p<0.01).</p