Alveolar ridge regeneration in two-wall-damaged extraction sockets of an in vivo experimental model.

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 < .05). CONCLUSIONS: Bone grafting with collagen membrane can maintain the alveolar ridge dimensions with substantial bone regeneration in a two-wall-damaged extraction socket.

Histologic analysis following grafting of damaged extraction sockets using deproteinized bovine or porcine bone mineral: A randomized clinical trial.

OBJECTIVES: This study histologically analyzed biopsy samples obtained from sites of damaged extraction socket grafting using deproteinized bovine bone mineral (DBBM) or deproteinized porcine bone mineral (DPBM) with coverage by a collagen membrane. MATERIAL AND METHODS: One hundred patients participated in this randomized controlled clinical trial of extraction socket grafts performed in cases of periodontally compromised teeth. All participants were blinded to their group allocations, and each material was grafted with coverage by collagen membranes after extraction of the tooth and removal of granulation tissue. At implant placement at 4 months, a biopsy was harvested at the implant site using a trephine was analyzed histologically. RESULTS: Eighty-five biopsy samples were acquired, of which 81 were finally included in the histologic analysis (42 in DBBM and 39 in DPBM group). Both DBBM and DPBM groups showed comparable proportions of residual biomaterial (12.37 ± 5.67% and 12.21 ± 5.75%, respectively), newly formed bone (15.07 ± 10.52% and 18.47 ± 11.47%, respectively), and nonmineralized tissue (72.56 ± 10.07% and 71.55 ± 15.47%, respectively). There were no significant differences in these histologic parameters between the two groups with different biomaterials. CONCLUSION: Comparable histologic bone formation was found in both socket grafted groups with DBBM or DPBM covered by collagen membranes in periodontally damaged extraction sockets. However, a wide variation in new bone formation was found after 4 months of postsurgical healing and a tendency of higher new bone formation was shown at damaged sockets that had an intact unilateral residual wall regardless of buccal or lingual side.

Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs.

Cellular reprogramming of somatic cells to patient-specific induced pluripotent stem cells (iPSCs) enables in vitro modelling of human genetic disorders for pathogenic investigations and therapeutic screens. However, using iPSC-derived cardiomyocytes (iPSC-CMs) to model an adult-onset heart disease remains challenging owing to the uncertainty regarding the ability of relatively immature iPSC-CMs to fully recapitulate adult disease phenotypes. Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) is an inherited heart disease characterized by pathological fatty infiltration and cardiomyocyte loss predominantly in the right ventricle, which is associated with life-threatening ventricular arrhythmias. Over 50% of affected individuals have desmosome gene mutations, most commonly in PKP2, encoding plakophilin-2 (ref. 9). The median age at presentation of ARVD/C is 26 years. We used previously published methods to generate iPSC lines from fibroblasts of two patients with ARVD/C and PKP2 mutations. Mutant PKP2 iPSC-CMs demonstrate abnormal plakoglobin nuclear translocation and decreased ß-catenin activity in cardiogenic conditions; yet, these abnormal features are insufficient to reproduce the pathological phenotypes of ARVD/C in standard cardiogenic conditions. Here we show that induction of adult-like metabolic energetics from an embryonic/glycolytic state and abnormal peroxisome proliferator-activated receptor gamma (PPAR-γ) activation underlie the pathogenesis of ARVD/C. By co-activating normal PPAR-alpha-dependent metabolism and abnormal PPAR-γ pathway in beating embryoid bodies (EBs) with defined media, we established an efficient ARVD/C in vitro model within 2 months. This model manifests exaggerated lipogenesis and apoptosis in mutant PKP2 iPSC-CMs. iPSC-CMs with a homozygous PKP2 mutation also had calcium-handling deficits. Our study is the first to demonstrate that induction of adult-like metabolism has a critical role in establishing an adult-onset disease model using patient-specific iPSCs. Using this model, we revealed crucial pathogenic insights that metabolic derangement in adult-like metabolic milieu underlies ARVD/C pathologies, enabling us to propose novel disease-modifying therapeutic strategies.

Clinical Factors and Cellular Responses of In Situ Human Alveolar Bone-Derived Mesenchymal Stromal Cells Associated With Early Periimplant Marginal Bone Loss: A Prospective Cohort Pilot Study.

PURPOSE: To investigate clinical factors and cellular responses of in situ human alveolar bone-derived mesenchymal stromal cells involved in early periimplant marginal bone loss. MATERIALS AND METHODS: Thirty-seven completely or partially edentulous patients were enrolled in this study. Periapical radiographs were taken at the time of implant surgery, at 3-month follow-up, and at 1-year follow-up. Univariate analysis and multiple logistic regression were performed to investigate the associations between marginal bone loss and study variables. The mRNA expression levels of 21 bone-remodeling- and tissue-healing-associated genes were analyzed by subgroup. RESULTS: Thirty-one patients with 98 implants were followed. The incidence and mean amount of bone loss were higher for overdentures than for other prosthesis and higher for the maxilla than for the mandible. The bone loss group showed lower mRNA expression levels of runt-related transcription factor-2, bone morphogenetic protein-2, and peroxisome proliferator-activated receptor gamma-2 and higher receptor activator of NKκB ligand/osteoprotegerin (RANKL/OPG) ratio. CONCLUSION: Within the limitations of the study, certain genes involved in bone remodeling (runt-related transcription factor-2 [Runx-2], bone morphogenetic protein-2 [BMP-2], and peroxisome proliferator-activated receptor gamma-2 [PPARγ-2]) and RANKL/OPG are correlated with early periimplant bone loss, with the type of suprastructure and the involved jaw being significant clinical factors.

Resonance Frequency Analysis of Tapered Implants Placed at Maxillary Posterior Sites After Lateral Sinus Augmentation: A 1.5-year Follow-Up Prospective Study.

OBJECTIVES: To use resonance frequency analysis to evaluate tapered implants placed at maxillary posterior sites after lateral sinus augmentation. MATERIALS AND METHODS: Patients who had missing teeth in the maxillary posterior area and required lateral sinus augmentation before implant placement were enrolled in this study. After a 6-month healing period, a tapered implant (Osstem TSIV) was placed. Implant success rate, survival rate, and marginal bone loss of the implants were measured. For resonance frequency analysis, implant stability quotient (ISQ) values were measured at each visit during a 1.5-year follow-up period. RESULTS: Twenty-four patients completed the study procedure. The residual bone height was 2.57 ± 1.10 mm (mean ± SD). Healing of the grafted area was uneventful in all cases, and 55 tapered implants were installed. The implant success rate was 95.56%, and the survival rate was 100% throughout the observation period. The marginal bone loss was limited to 0.22 ± 0.44 mm. ISQ increased gradually from 68.40 ± 11.14 to 82.24 ± 4.75 during the 1.5-year follow-up period. CONCLUSION: The tapered implants showed good initial and final stability after placement in the soft bone of the maxillary posterior area after lateral sinus augmentation.

Analysis of fatty acids in mouse tissue via in situ transmethylation with biochar.

Lipid derivatization technology-mediated fatty acid profiling studies have been suggested to dissect the contents of lipids in white fat and brown fat tissue. The focus of this study is to profile fatty acid lipidomics in brown adipose tissue and white adipose tissue of mice by derivatizing their lipids into fatty acid methyl esters via in situ transmethylation using a rice husk-derived biochar as porous media. The in situ transmethylation using biochar is advantageous in biological analysis because there was no loss of samples inevitably occurring in the loss of lipid in solvent extraction and purification steps.

Characteristics of Local Delivery Agents for Treating Peri-Implantitis on Dental Implant Surfaces: A Preclinical Study.

Local delivery agents (LDAs) are widely used in peri-implantitis treatments. The aim of this study was to identify LDAs remaining on the dental implant surfaces and to analyze the components of these residues after applying various cleaning methods. Implants were prepared with a sand-blasted, large-grit, acid-etched surface. Four kinds of LDAs were applied on the implant surfaces: chlorhexidine gel (group 2), tetracycline solution (group 3), and 2 kinds of minocycline hydrochloride agents (groups 4 and 5). Group 1 received normal saline as a control. Two cleaning methods were applied for different durations as follows: (1) running distilled water for 10 seconds (subgroup A), 5 minutes (subgroup B), and 15 minutes (subgroup C); and (2) water spray of a dental-unit chair for 10 seconds (subgroup D) and 5 minutes (subgroup E). Scanning electron microscopy and energy-dispersive x-ray spectroscopy were used to analyze the surface morphology and residue components for all implants. The amount of LDA removed from the implant surfaces in groups 1, 2, 3, and 5 increased with the cleaning duration and pressure. However, Minocline remained coated on the implant surfaces in group 4 under all cleaning conditions. Minocline could not be cleaned off well by water due to its hydrophobicity. Therefore, directly using this agent on implant surfaces with peri-implantitis should be carefully considered. The presence of LDA residues without drug efficacies on implant surfaces might interfere with reosseointegration and act as a reservoir of microorganisms.

Radiographic and histologic observations of sequential healing processes following ridge augmentation after tooth extraction in buccal-bone-deficient extraction sockets in beagle dogs.

OBJECTIVES: To evaluate dimensional ridge alterations and sequential healing processes following ridge augmentation after tooth extraction in damaged extraction sockets with buccal-bone-deficiency. MATERIAL AND METHODS: Bilateral dental roots of three mandibular premolars were extracted with entire removal of the buccal-bone plate in eight beagle dogs. Unilateral sites were grafted with biomaterials (test group) and contralateral sites were healed without grafting (control group). Observations were made after 1, 2, 4, and 8 weeks, and all sites were distributed evenly (n = 6 for each group and period). Radiographic/histomorphometric analyses were performed. RESULTS: In spontaneous healing of damaged extraction sockets, the dimension of regenerated alveolar ridge gradually increased until 4 weeks and then remained stable, but radiographic/histomorphometric analyses revealed evident dimensional shrinkage compared to the pristine tissue at 8 weeks in the coronal and middle areas. Bone grafting retained the pristine dimension of alveolar ridge, and newly formed bone area within the augmented space continuously expanded during the observational period to the outermost border of the space. CONCLUSIONS: Spontaneous healing of damaged extraction sockets caused substantial dimensional shrinkage. However, ridge augmentation can provide space into which new bone may grow continuously, resulting in the final dimensions comparable to those of the pristine alveolar ridge.

Chemokine in inflamed periodontal tissues activates healthy periodontal-ligament stem cell migration.

AIM: The present study aimed to characterize the expression pattern of chemokines obtained from inflamed periodontal defects and to determine the characteristics of human periodontal-ligament stem cells (hPDLSCs) migrated by each specific chemokine. MATERIALS AND METHODS: Both inflamed and healthy periodontal tissues were obtained from periodontitis patients (n = 11), and the chemokine expression levels were analyzed. The periodontal-tissue-specific chemokines were applied to healthy hPDLSCs from extracted teeth (n = 3), with FGF-2 acting as a positive control. Cells were separated by selected chemokines using transwell method into migrated/unmigrated hPDLSCs. The characteristics of the hPDLSC subpopulation recruited by each chemokine were assessed, and gene expression pattern was analyzed by microarray. RESULTS: Chemokines were categorized into three groups by specific patterns of "appearing," "increasing," and "decreasing/disappearing" from healthy to inflamed tissues. A representative chemokine from each group enhanced the capacities for colony formation and osteogenic/adipogenic differentiation while maintaining the surface markers of hPDLSCs. RANTES/CCL5 significantly increased the cellular migration of hPDLSCs, via enhancement of signaling pathways, regulation of the actin skeleton, and focal adhesion. CONCLUSION: The present study found a specific chemokine profile induced by inflammation in periodontal tissues, with RANTES/CCL5 appearing to play a role in the migration of hPDLSCs into inflammatory periodontal lesions.

In Vivo Evaluation of Commercially Available Gel-Type Polyethylene Glycol Membrane for Carrier of Recombinant Human Bone Morphogenetic Protein-2.

PURPOSE: This study evaluated a commercially available, 3-dimensional gel-type polyethylene glycol (PEG) membrane as a carrier for recombinant human bone morphogenetic protein-2 (rhBMP-2) using a rat calvarial defect model. Another gel-type carrier, fibrin-fibronectin system (FFS), was used as a positive control. MATERIALS AND METHODS: Critical-size defects were made in the rat calvarium, which were allocated to 1 of 10 groups comprising 2 healing periods and biomaterial conditions: 1) sham control, 2) FFS only, 3) FFS plus BMP-2, 4) PEG only, and 5) PEG plus BMP-2. Radiographic and histologic analyses were performed at 2 and 8 weeks after surgery. RESULTS: After 2 weeks, some parts of the FFS were biodegraded and extensive cellular infiltration was observed at sites that received FFS or FFS plus BMP-2. The PEG membrane retained its augmented volume without cellular infiltration at sites that received PEG or PEG plus BMP-2. After 8 weeks, the FFS was completely degraded and replaced by new bone and connective tissues. In contrast, the volume of residual PEG was similar to that at 2 weeks, with slight cellular infiltration. In particular, there was progressive bone regeneration around micro-cracks and resorbed outer surface in the PEG + BMP-2 group. Although the PEG + BMP-2 group showed increased area and percentage of new bone, there was no statistical relevance after 2 and 8 weeks in histomorphometric analyses. However, the appearance of the healing differed (with new bone formation along micro-cracks in the PEG + BMP-2 group), and further studies with longer healing periods are needed to draw conclusions about clinical applications. CONCLUSION: Evidence of mechanical stability and new bone formation along micro-cracks when using PEG plus BMP-2 might support the PEG membrane as a candidate carrier material for rhBMP-2.

Autonomous and nonautonomous regulation of Wnt-mediated neuronal polarity by the C. elegans Ror kinase CAM-1.

Wnts are a conserved family of secreted glycoproteins that regulate various developmental processes in metazoans. Three of the five Caenorhabditis elegans Wnts, CWN-1, CWN-2 and EGL-20, and the sole Wnt receptor of the Ror kinase family, CAM-1, are known to regulate the anterior polarization of the mechanosensory neuron ALM. Here we show that CAM-1 and the Frizzled receptor MOM-5 act in parallel pathways to control ALM polarity. We also show that CAM-1 has two functions in this process: an autonomous signaling function that promotes anterior polarization and a nonautonomous Wnt-antagonistic function that inhibits anterior polarization. These antagonistic activities can account for the weak ALM phenotypes displayed by cam-1 mutants. Our observations suggest that CAM-1 could function as a Wnt receptor in many developmental processes, but the analysis of cam-1 mutants may fail to reveal CAM-1's role as a receptor in these processes because of its Wnt-antagonistic activity. In this model, loss of CAM-1 results in increased levels of Wnts that act through other Wnt receptors, masking CAM-1's autonomous role as a Wnt receptor.

Missense mutations in plakophilin-2 cause sodium current deficit and associate with a Brugada syndrome phenotype.

BACKGROUND: Brugada syndrome (BrS) primarily associates with the loss of sodium channel function. Previous studies showed features consistent with sodium current (INa) deficit in patients carrying desmosomal mutations, diagnosed with arrhythmogenic cardiomyopathy (or arrhythmogenic right ventricular cardiomyopathy). Experimental models showed correlation between the loss of expression of desmosomal protein plakophilin-2 (PKP2) and reduced INa. We hypothesized that PKP2 variants that reduce INa could yield a BrS phenotype, even without overt structural features characteristic of arrhythmogenic right ventricular cardiomyopathy. METHODS AND RESULTS: We searched for PKP2 variants in the genomic DNA of 200 patients with a BrS diagnosis, no signs of arrhythmogenic cardiomyopathy, and no mutations in BrS-related genes SCN5A, CACNa1c, GPD1L, and MOG1. We identified 5 cases of single amino acid substitutions. Mutations were tested in HL-1-derived cells endogenously expressing NaV1.5 but made deficient in PKP2 (PKP2-KD). Loss of PKP2 caused decreased INa and NaV1.5 at the site of cell contact. These deficits were restored by the transfection of wild-type PKP2, but not of BrS-related PKP2 mutants. Human induced pluripotent stem cell cardiomyocytes from a patient with a PKP2 deficit showed drastically reduced INa. The deficit was restored by transfection of wild type, but not BrS-related PKP2. Super-resolution microscopy in murine PKP2-deficient cardiomyocytes related INa deficiency to the reduced number of channels at the intercalated disc and increased separation of microtubules from the cell end. CONCLUSIONS: This is the first systematic retrospective analysis of a patient group to define the coexistence of sodium channelopathy and genetic PKP2 variations. PKP2 mutations may be a molecular substrate leading to the diagnosis of BrS.

Ridge regeneration of damaged extraction sockets using rhBMP-2: an experimental study in canine.

OBJECTIVES: This study evaluated the dimensional ridge alteration in a buccal-bone-deficient extraction socket, and ridge regeneration following socket grafting accompanied by recombinant human bone morphogenetic protein 2 (rhBMP-2) or a collagen membrane covering. MATERIAL AND METHODS: In five beagle dogs, entire buccal bone of the extracted sockets of premolars was surgically removed and immediately grafted using one of the following graft protocols: (1) sham surgery without any grafting, and grafting with (2) deproteinized bovine bone mineral (DBBM), (3) DBBM/rhBMP-2 and (4) DBBM covered with a collagen membrane (DBBM/Membrane). Quantitative/qualitative analyses were performed radiographically/histologically after 8 weeks. RESULTS: Buccal-deficient extraction sockets healed with significant reduction in buccolingual dimension along the entire length of the socket, but all grafting techniques reduced the dimensional changes compared to the non-grafted control sites. Histologically, sites received DBBM only exhibited minimal regeneration, whereas sites grafted with DBBM/rhBMP-2 or DBBM/Membrane exhibited greater new bone formation extending the entire augmented area. CONCLUSIONS: Buccal-bone-deficiency may lead to significant volume reduction after tooth extraction along the entire length of the socket, and socket grafting accompanied by rhBMP-2 or covered with a membrane can be candidate therapies for preservation of the buccolingual dimension and successful ridge regeneration.

Apatite-coated collagen sponge for the delivery of bone morphogenetic protein-2 in rabbit posterolateral lumbar fusion.

Bone morphogenetic proteins (BMPs) need an effective delivery system for efficient bone regeneration. In this study, we evaluated the efficiency of an apatite-coated collagen sponge for the long-term delivery of BMP-2 in a rabbit model of lumbar posterolateral fusion. A total of 15 rabbits, divided into three groups, underwent posterolateral lumbar fusion. The first group (control group) received uncoated collagen sponges without BMP-2. The second group (uncoated group) received uncoated collagen sponges with BMP-2 (40 µg each side). The third group (apatite-coated group) received apatite-coated collagen sponges with the same level of BMPs (40 µg each side). All rabbits were euthanized 6 weeks after operation, and the fusion status was assessed by radiographic study, micro-CT, manual palpation, biomechanical study, and histological examination. Fusion rates as determined by radiographic study, micro-CT, and manual palpation showed that the apatite-coated group had a significantly higher rate of fusion than the control group (P = 0.024), while the uncoated group did not (P = 0.083). Biomechanical study showed significantly higher tensile strength in the apatite-coated group than the uncoated group (P = 0.032). Denser trabeculations were found in the apatite-coated group compared with the uncoated group. It is concluded that the use of apatite-coated collagen sponges for BMP-2 delivery enhanced bone regeneration.

Influence of nanocoated calcium phosphate on two different types of implant surfaces in different bone environment: an animal study.

OBJECTIVE: The purpose of this study was to evaluate the osseointegration of two different types of surfaces, smooth and roughened surface implants nanocoated with calcium phosphate (CAP) around different bone environment. MATERIALS AND METHODS: Five male mongrel dogs were used in this study. The premolars and molars were extracted on both sides of the mandible. Eight weeks after extraction, implants were submerged on both sides of the mandible. On the left, CAP nanocoated roughened surface (RCAP) implants were installed whereas, the CAP nanocoated smooth surface (SCAP) implants were installed on the right side. The control group had no defect, on the other hand, three-wall intrabony defects were surgically created adjacent to the implant in the experimental group. The dogs were sacrificed after 12 weeks. RESULTS: Histological and histomorphometrical analysis were performed with the specimen. The SCAP and RCAP implants showed good osseointegration with no statistical significance in the control group. Histologically, the SCAP group showed little resolution of the defect compared with the RCAP group. In the experimental groups, there was a significant difference in defect fill between SCAP and RCAP. CONCLUSION: Within the limits of our study, it can be concluded that SCAP and RCAP implants show no difference in sufficient bone area whereas, CAP nanocoating on roughened implant surface may enhance osseointegration in deficient bone environment.

Long-term retrospective study of narrow implants for fixed dental prostheses.

OBJECTIVE: The objectives of this study were to analyze retrospectively the long-term survival and success rates of Narrow implants (NIs) placed with various implant systems, and the association with biological and technical complications. MATERIAL AND METHODS: In total, 338 patients (men = 45.6%, women = 54.4%) who received 541 NIs (≤3.5 mm in diameter) for fixed prostheses were enrolled in this retrospective study. The mean marginal bone level (MMBL) change was calculated. Life table analysis with the cumulative survival rate and success rate was calculated, and biological and technical complications were evaluated. RESULTS: The annual MMBL change was 0.07 ± 0.20 mm. The 12-year cumulative survival (success) rates of NIs were 98.1% (91.8%) and 98.5% (93.8%) for the implant- and subject-based analysis, respectively. During the observation period up to 12 years (mean 4.9 years), six implants were lost in the maxilla, whereas three implants were lost in the mandible. Technical complications were more frequent than biological complications. Infection was the most common underlying cause of biological complications and the most frequent technical complication was decementation. CONCLUSION: In conclusion, the findings of this study suggest that NIs could be used safely for narrow alveolar ridges or narrow mesiodistal spaces on the basis of their high survival rate.

Controlled delivery of low-dose bone morphogenetic protein-2 using heparin-conjugated fibrin in the posterolateral lumbar fusion of rabbits.

The heparin-conjugated fibrin (HCF) system has been developed to deliver bone morphogenetic proteins (BMPs) for a long-term period and thus enhance bone regeneration. In the present study, we tested the effectiveness of the delivery system for spinal fusion with a very low dose of BMP-2. A total of 15 rabbits underwent posterolateral lumbar spine, divided into three groups. The control group received only collagen sponges without BMP-2, another group (BMP-only group) received collagen sponges loaded with BMP-2 (10 µg each side), and the last group (BMP/HCF group) received collagen sponges filled with HCF loaded with BMP-2 (10 µg each side). All animals were euthanized 8 weeks after surgery, and the fusion was assessed by radiographs, manual palpation, computed tomography scan, and mechanical testing. No case in the BMP/HCF group or in the control group achieved solid fusion, while all cases in BMP-only group showed evidence of solid fusion. BMP/HCF group had significantly lower fusion rate and tensile strength than BMP-only group at the dose of 10 µg of BMP-2. The HCF long-term delivery system with the low dose of BMP-2 (10 µg) is ineffective for the induction of lumbar posterolateral fusion in the rabbits.

Comparing the clinical effects of local administration of minocycline ointment and doxycycline solution in the treatment of acute periodontal abscesses: a retrospective clinical study.

PURPOSE: The objective of this retrospective clinical study was to provide evidence supporting the adjunctive local application of doxycycline solution or minocycline ointment, in conjunction with drainage, for the treatment of acute periodontal abscesses. METHODS: The study included 63 patients who had received treatment for acute periodontal abscesses through drainage supplemented with 1 of 3 types of adjunctive medications during their initial visit (visit 1; baseline): 1) saline irrigation (the control group), 2) 2% minocycline ointment (the TM group), or 3) 300 mg/mL doxycycline irrigation (the TD group). The same adjunctive medication was administered at visit 2, which took place 1 week after visit 1. Probing depth (PD), bleeding on probing (BOP), plaque index, gingival recession, clinical attachment level, and tooth mobility were clinically evaluated at visits 1, 2, and a third visit (visit 3; 4 weeks after visit 1). Statistical significance was considered to be indicated by P values <0.05. RESULTS: By visit 3, all clinical indices and tooth mobility had significantly decreased in each group. At this visit, PD and BOP on the abscess side were significantly lower in the TM and TD groups compared to the control group. The TD group showed a significantly greater improvement than the TM group, with mean PD reductions of 1.09 mm in the control group, 1.88 mm in the TM group, and 2.88 mm in the TD group. Similarly, mean BOP reductions were 45% in the control group, 73.02% in the TM group, and 95.45% in the TD group. CONCLUSIONS: Local and adjunctive administration of doxycycline and minocycline in combination with drainage exhibited clinical advantages over drainage alone in improving PD and BOP. Notably, a doxycycline solution of 300 mg/mL was more effective than a 2% minocycline ointment.

Injectable contraceptive Depo-Provera induces erratic beating patterns in patient-specific induced pluripotent stem cell-derived cardiomyocytes with long QT syndrome type 2.

BACKGROUND: Long QT syndrome type 2 (LQT2) is caused by pathogenic variants in KCNH2. LQT2 may manifest as QT prolongation on an electrocardiogram and present with arrhythmic syncope/seizures and sudden cardiac arrest/death. Progestin-based oral contraceptives may increase the risk of LQT2-triggered cardiac events in women. We previously reported on a woman with LQT2 and recurrent cardiac events temporally related and attributed to the progestin-based contraceptive medroxyprogesterone acetate ("Depo-Provera" [Depo] MilliporeSigma, Catalog# 1378001, St. Louis, MO). OBJECTIVE: The purpose of this study was to evaluate the arrhythmic risk of Depo in a patient-specific induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM) model of LQT2. METHODS: An iPSC-CM line was generated from a 40-year-old woman with p.G1006Afs∗49-KCNH2. A CRISPR/Cas9 gene-edited/variant-corrected isogenic control iPSC-CM line was generated. FluoVolt (Invitrogen, F10488, Waltham, MA) was used to measure the action potential duration after treatment with 10 µM Depo. Erratic beating patterns characterized as alternating spike amplitudes, alternans, or early afterdepolarization-like phenomena were assessed using multielectrode array (MEA) after 10 µM Depo, 1 µM isoproterenol (ISO), or combined Depo + ISO treatment. RESULTS: Depo treatment shortened the action potential duration at 90% repolarization of G1006Afs∗49 iPSC-CMs from 394 ± 10 to 303 ± 10 ms (P < .0001). Combined Depo + ISO treatment increased the percentage of electrodes displaying erratic beating in G1006Afs∗49 iPSC-CMs (baseline: 18% ± 5% vs Depo + ISO: 54% ± 5%; P < .0001) but not in isogenic control iPSC-CMs (baseline: 0% ± 0% vs Depo + ISO: 10% ± 3%; P = .9659). CONCLUSION: This cell study provides a potential mechanism for the patient's clinically documented Depo-associated episodes of recurrent ventricular fibrillation. This in vitro data should prompt a large-scale clinical assessment of Depo's potential proarrhythmic effect in women with LQT2.

Electrophysiological mapping of embryonic mouse hearts: mechanisms for developmental pacemaker switch and internodal conduction pathway.

INTRODUCTION: Understanding sinoatrial node (SAN) development could help in developing therapies for SAN dysfunction. However, electrophysiological investigation of SAN development remains difficult because mutant mice with SAN dysfunctions are frequently embryonically lethal. Most research on SAN development is therefore limited to immunocytochemical observations without comparable functional studies. METHODS AND RESULTS: We applied a multielectrode array (MEA) recording system to study SAN development in mouse hearts acutely isolated at embryonic ages (E) 8.5-12.5 days. Physiological heart rates were routinely restored, enabling accurate functional assessment of SAN development. We found that dominant pacemaking activity originated from the left inflow tract (LIFT) region at E8.5, but switched to the right SAN by E12.5. Combining MEA recordings and pharmacological agents, we show that intracellular calcium (Ca(2+))-mediated automaticity develops early and is the major mechanism of pulse generation in the LIFT of E8.5 hearts. Later in development at E12.5, sarcolemmal ion channels develop in the SAN at a time when pacemaker channels are down-regulated in the LIFT, leading to a switch in the dominant pacemaker location. Additionally, low micromolar concentrations of tetrodotoxin (TTX), a sodium channel blocker, minimally affect pacemaker rhythm at E8.5-E12.5, but suppress atrial activation and reveal a TTX-resistant SAN-atrioventricular node (internodal) pathway that mediates internodal conduction in E12.5 hearts. CONCLUSIONS: Using a physiological mapping method, we demonstrate that differential mechanistic development of automaticity between the left and right inflow tract regions confers the pacemaker location switch. Moreover, a TTX-resistant pathway mediates preferential internodal conduction in E12.5 mouse hearts.