Correction: Influence of the shared epitope on the elicitation of experimental autoimmune arthritis biomarkers.

[This corrects the article DOI: 10.1371/journal.pone.0250177.].

Influence of the shared epitope on the elicitation of experimental autoimmune arthritis biomarkers.

Our previous studies have shown that inoculation of the oral cavity of "humanized" B6.DR1/4 mice with the periodontal pathogen Porphyromonas gingivalis results in an increase in the percentage of circulating Th17 cells, loss of bone and an exacerbation of experimental autoimmune arthritis. The aim of this study was to assess the role played by the human HLA-DRß molecule containing the shared epitope supplied as a transgene to I-A˚ (murine class II null) C57BL/6 (B6) mice in driving these findings. We compared various immune response parameters as well as alveolar and peri-articular bone loss between humanized B6.DR1 (or B6.DR4) mice and their WT (B6) counterparts. We found that the presence of the shared epitope in the context of inoculation with P. gingivalis enhanced the percentage of Th17 cells generated, dramatically enhanced bone loss and importantly allowed for the generation of CCP2⁺ ACPAs that are not found in C57BL/6 or DBA/1 arthritic mouse serum. Due to the exceedingly complex nature of environmental factors impacting on genetic elements, it has been difficult to unravel mechanisms that drive autoimmune arthritis in susceptible individuals. The findings in this study may provide one small piece of this puzzle that can help us to better understand part of this complexity.

Understanding Early-Stage Posttraumatic Osteoarthritis for Future Prospects of Diagnosis: from Knee to Temporomandibular Joint.

PURPOSE OF REVIEW: Many mechanical load-bearing joints of the body are prone to posttraumatic osteoarthritis (PTOA), including the knee joint and temporomandibular joint (TMJ). Early detection of PTOA can be beneficial in prevention or alleviating further progression of the disease. RECENT FINDINGS: Various mouse models, similar to those used in development of novel diagnosis strategies for early stages of OA, have been proposed to study early PTOA. While many studies have focused on OA and PTOA in the knee joint, early diagnostic methods for OA and PTOA of the TMJ are still not well established. Previously, we showed that fluorescent near-infrared imaging can diagnose inflammation and cartilage damage in mouse models of knee PTOA. Here we propose that the same approach can be used for early diagnosis of TMJ-PTOA. In this review, we present a brief overview of PTOA, application of relevant mouse models, current imaging methods available to examine TMJ-PTOA, and the prospects of near-infrared optical imaging to diagnose early-stage TMJ-OA.

A comparison of two types of electrospun chitosan membranes and a collagen membrane in vivo.

BACKGROUND: Electrospun chitosan membranes subjected to post-spinning processes using either triethylamine/tert-butyloxycarbonyl (TEA/tBOC) or butyryl-anhydride (BA) modifications to maintain nanofiber structure have exhibited potential for use in guided bone regeneration applications. The aim of this study was to evaluate ability of the modified membranes to support healing of bone-grafted defects as compared to a commercial collagen membrane. METHOD: TEA/tBOC-treated and BA-treated chitosan membranes were characterized for fiber morphology by electron microscopy, residual trifluoroacetic acid by19F NMR and endotoxin level using an endotoxin quantitation kit (ThermoScientific, US). Chitosan membranes were cut into 12 mm diameter disks. An 8 mm calvarial defect was created in each of 48 male rats and then filled with Bio-Oss (Geistlich, US) bone graft. The grafted defects were covered with either (1) TEA/tBOC-treated chitosan membrane (2) BA-treated chitosan membrane or (3) the control BioMend Extend (Zimmer Biomet, US) collagen membrane. After 3 and 8 weeks, the rats were euthanized and calvaria was retrieved for microCT and histological analyses (n = 8/group/time points). RESULTS: Both TEA/tBOC-treated and BA-treated membranes were composed of nanofibers in the ∼231 to ∼284 nm range respectively, exhibited no TFA salt residue and low endotoxin levels (≤0.1 ± 0.01 EU/membrane). All membranes supported increased bone growth from 3 weeks to 8 weeks though there was no significant difference among the membrane types. However, TEA/tBOC treated and BA treated chitosan membranes both showed significantly greater bone density (∼6% greater at 3 weeks and ∼8% greater at 8 weeks) as compared to BioMend Extend collagen membrane at both time points (p = 0.0002). CONCLUSIONS: Chitosan membranes supported better bone healing based on bone density than the collagen membrane.

Vitamin D attenuates human gingival fibroblast inflammatory cytokine production following advanced glycation end product interaction with receptors for AGE.

BACKGROUND AND OBJECTIVES: Vitamin D [1,25(OH)2 D3 or 1,25D3] is critical in musculoskeletal health, inflammation, immune response, and glucose metabolism. Patients with vitamin D deficiency may be at higher risk of diabetes and periodontitis. Diabetic patients exhibit exacerbated inflammation and more periodontal destruction. Advanced glycation end products (AGEs), formed during diabetic hyperglycemia, activate inflammatory pathways in periodontitis. Human gingival fibroblasts (HGFs) express receptors for AGEs (RAGEs) and can contribute to inflammation. OBJECTIVES: Determine whether glycated human serum albumin (G-HSA) augments HGF IL-6 and IL-8 production, and whether treatment with 1,25D3 attenuates cytokine production following stimulation with G-HSA + IL-1ß and/or IL-17. MATERIAL AND METHODS: HGFs were incubated ±G-HSA or normal human serum albumin (HSA), ±IL-1ß and/or IL-17, ±1,25D3. Cytokines were measured by ELISA. Neutralizing anti-RAGE was used to assess AGE-RAGE interaction. Endotoxin was measured using the ToxinSensor™ System. Data were expressed as mean ± standard deviation and analyzed using a one-way analysis of variance (ANOVA) and Scheffe's F procedure for post hoc comparisons. RESULTS: G-HSA or IL-1ß, but not HSA, significantly stimulated IL-6 and IL-8 production. G-HSA or HSA when combined with IL-1ß or IL-1ß + IL-17 synergistically stimulated IL-6 and IL-8. Neutralizing anti-RAGE inhibited IL-6 and IL-8 produced by cells stimulated with IL-1ß + G-HSA but not (+HSA). Synergism caused by HSA did not appear to be mediated by endotoxin since its levels in G-HSA and HSA were not sufficient to stimulate fibroblasts. Vitamin D inhibited IL-6 and IL-8 production stimulated by G-HSA or HSA + IL-1ß or IL-1ß + IL-17. CONCLUSIONS: Results suggest that the "perioprotective" effects of vitamin D are related to its ability to regulate inflammatory cytokine production by HGFs following AGE-RAGE interaction.

Mechanically stable surface-hydrophobilized chitosan nanofibrous barrier membranes for guided bone regeneration.

The use of chitosan based nanofiber membranes in guided bone regeneration (GBR) is limited by its uncontrolled swelling and mechanical instability in aqueous environments. This paper describes the significantly improved stability and properties of surface butyrylated chitosan nanofiber (BCSNF) membranes that greatly enhance their potential in GBR. The BCSNF membranes exhibited an overall degree of substitution of 1.61, an average diameter of 99.3 ± 33.7 nm, and a 75% decrease in swelling with an approximate doubling in suture pull out strengths as compared to unmodified fibers in aqueous environment. In a five week phosphate-buffered saline-lysozyme degradation study, it was found that the remaining mass fraction of BCSNF membranes was 11.5% more than that of unmodified fibers. In vitro, the BCSNF membranes were found to support the adhesion and proliferation of fibroblasts and were cell occulusive. In vivo, the BCSNF membranes were found to significantly improve the regeneration of a rat calvarial critical size defect in a 12 week healing period and showed better barrier function than commercially available collagen membranes with little soft tissue penetration through the membranes. Taken together, these data provide strong scientific evidence for use of BCSNF membranes in GBR applications.

Dental and Dental Hygiene Intraprofessional Education: A Pilot Program and Assessment of Students' and Patients' Satisfaction.

Interprofessional and intraprofessional education (when students from two or more professions or within the same profession, respectively, learn about, from, and/or with each other) is crucial for effective interdisciplinary collaboration. The aims of this study were to assess the effectiveness of a clinical intraprofessional education program for dental and dental hygiene students, based on students' expectations and satisfaction with the program and patients' satisfaction with the team-based care. The pilot program was developed at the University of Tennessee Health Science Center College of Dentistry, where dental hygiene students were paired randomly with dental students scheduled for prophylaxis, scaling and root planing, or periodontal maintenance. Surveys with questions about the students' expectations and satisfaction were distributed to 89 senior dental students and 27 senior dental hygiene students before and after team-based procedures. Another survey was distributed to 17 patients asking about their satisfaction with the team-based care. All 27 dental hygiene students (100% response rate), 51 dental students (57.3% response rate), and all 17 patients (100% response rate) participated in the surveys. The results showed that both the dental and dental hygiene students had high expectations and were overall satisfied with the intraprofessional education. The students' expectations and perceived educational gap (difference between expectations and satisfaction) differed for the dental and dental hygiene students (p<0.001). The male dental students were also more satisfied than the female dental students (p<0.01). Overall, the program met or exceeded the students' expectations, and the patients were overwhelmingly satisfied with the team-based care. These results suggest that this intraprofessional practice model provided an effective educational experience for both dental and dental hygiene students and patients. The differences between the dental hygiene and dental students' expectations will help in the design of more effective training that promotes intraprofessional and interprofessional teamwork.

In vitro and in vivo evaluations of a novel post-electrospinning treatment to improve the fibrous structure of chitosan membranes for guided bone regeneration.

Electrospun chitosan membranes have been investigated for guided bone regeneration but are susceptible to swelling, dissolution, and loss of biomimetic nanofiber structure due to residual acid salts. A novel process was investigated for acidic salt removal from chitosan electrospun in 70% trifluoroacetic acid (TFA) by treating with triethylamine (TEA)/acetone and di-tert-butyl dicarbonate (tBOC) instead of the common Na2CO3 treatment. TFA salt removal and nanofiber structure stabilization were confirmed by EDS, FTIR, 19F NMR and electron microscopy before and after soaking in water. Membrane degradation after 4 weeks in PBS with 100 µg ml-1 lysozyme and osteoblastic proliferation were similar between TEA/tBOC-treated and Na2CO3-treated membranes. A simulated surgical tear test using surgical tacks showed that the peak tensile tear strength of the TEA/tBOC-treated chitosan membranes (62.1 ± 1.9 N mm-1) was significantly greater than a commercial polylactic acid (PLA) membrane (13.4 ± 0.4 N mm-1), similar to one commercial collagen membrane (55.3 ± 7.5 N mm-1) but lower than another commercial collagen membrane (133.9 ± 21.5 N mm-1). Rat 8 mm critical-sized calvarial defects covered with TEA/tBOC-treated chitosan membranes prevented soft tissue infiltration and supported new bone growth (15.76 ± 10.28%) similar to a commercial collagen membrane (16.08 ± 10.69%) at 12 weeks based on microCT analyses. Hence our novel TEA/tBOC process significantly improved nanofiber structure and mechanical strengths of electrospun chitosan membranes as compared to Na2CO3 treated membranes, without affecting in vitro degradation or cytocompatibility, improved membrane mechanical properties to be greater than a commercial PLA membrane and to be in range of commercial collagen membranes and supported calvarial bone defect healing similar to collagen. Thus TEA/tBOC-treated chitosan membranes exhibit many characteristics and properties that strongly support their potential for use in guided bone regeneration.

Bone loss and aggravated autoimmune arthritis in HLA-DRß1-bearing humanized mice following oral challenge with Porphyromonas gingivalis.

BACKGROUND: The linkage between periodontal disease and rheumatoid arthritis is well established. Commonalities among the two are that both are chronic inflammatory diseases characterized by bone loss, an association with the shared epitope susceptibility allele, and anti-citrullinated protein antibodies. METHODS: To explore immune mechanisms that may connect the two seemingly disparate disorders, we measured host immune responses including T-cell phenotype and anti-citrullinated protein antibody production in human leukocyte antigen (HLA)-DR1 humanized C57BL/6 mice following exposure to the Gram-negative anaerobic periodontal disease pathogen Porphyromonas gingivalis. We measured autoimmune arthritis disease expression in mice exposed to P. gingivalis, and also in arthritis-resistant mice by flow cytometry and multiplex cytokine-linked and enzyme-linked immunosorbent assays. We also measured femoral bone density by microcomputed tomography and systemic cytokine production. RESULTS: Exposure of the gingiva of DR1 mice to P. gingivalis results in a transient increase in the percentage of Th17 cells, both in peripheral blood and cervical lymph nodes, a burst of systemic cytokine activity, a loss in femoral bone density, and the generation of anti-citrullinated protein antibodies. Importantly, these antibodies are not produced in response to P. gingivalis treatment of wild-type C57BL/6 mice, and P. gingivalis exposure triggered expression of arthritis in arthritis-resistant mice. CONCLUSIONS: Exposure of gingival tissues to P. gingivalis has systemic effects that can result in disease pathology in tissues that are spatially removed from the initial site of infection, providing evidence for systemic effects of this periodontal pathogen. The elicitation of anti-citrullinated protein antibodies in an HLA-DR1-restricted fashion by mice exposed to P. gingivalis provides support for the role of the shared epitope in both periodontal disease and rheumatoid arthritis. The ability of P. gingivalis to induce disease expression in arthritis-resistant mice provides support for the idea that periodontal infection may be able to trigger autoimmunity if other disease-eliciting factors are already present.

Dentoskeletal effects of a temporary skeletal anchorage device-supported rapid maxillary expansion appliance (TSADRME): A pilot study.

OBJECTIVE: To quantitatively evaluate maxillary skeletal expansion using cone-beam computed tomography (CBCT) images and propose a novel way to quantify the dental tipping effects of temporary skeletal anchorage device-supported rapid maxillary expansion appliance (TSADRME). MATERIALS AND METHODS: Images from 25 patients receiving rapid maxillary expansion with incorporated temporary skeletal anchorage devices (TSADs) before activation (T1) and after removal (T2) were analyzed to detect dentoskeletal changes. RESULTS: A significant increase from T1 to T2 was found for all linear measurements except buccal maxillary width at the canines. The greatest buccal expansion was at the first molar, decreasing anteriorly. However, the greatest palatal expansion was at the first premolar. All younger subjects (8-16 years old) exhibited less dental tipping and greater expansion overall compared with the older subjects. There was great variability in dental tipping of first molars (mean = 4.31°), with some subjects demonstrating mild uprighting of these teeth. CONCLUSIONS: The TSADRME appliance is an effective, clinically useful device that results in mild molar tipping and may positively affect expansion in the area of TSAD placement.

Experience and expertise regarding orthodontic management of childhood and adolescent cancer survivors.

INTRODUCTION: Great strides in pediatric cancer treatment are allowing hundreds of thousands of children to survive into adulthood. However, these treatments can be responsible for long-term medical and dental complications. The treatments may alter the patients' dental health and require modifications to standard orthodontic care. The aim of this study was to examine knowledge and clinical experience regarding orthodontic management of childhood cancer survivors. METHODS: A 12-question online survey consisting of 3 sections was sent to 2500 randomly selected members of the American Association of Orthodontists and all 2300 members of the Southern Association of Orthodontists. The first section consisted of questions about the respondents' practice characteristics, the second questioned how many survivor patients the respondent had treated, and the third included questions about specific (anonymous) patient experiences and treatment modifications. RESULTS AND CONCLUSIONS: There were 381 responses. The data from this study suggest a tendency for more experienced practitioners to have treated survivors of childhood cancer. Orthodontic education regarding the treatment of these patients is limited. Although most orthodontists reported having treated such patients, few had treated more than 10. There is a need for more information regarding dental complications of pediatric cancer treatment and for guidelines for the orthodontic treatment of these patients.

Management of oral melanin pigmentation.

Melanin is an endogenous pigment responsible for human tissue coloration of the skin, mucosa, hair, eyes and parts of the brain. In the skin, its function is protection from the harmful effects of UV radiation. Its purpose in oral tissues has not yet been determined. Oral pigmentation could be an esthetic issue for some patients, particularly when it is located on the anterior labial gingiva in individuals with a high smile line. This article presents and describes several different approaches for the management of oral melanin pigmentation.

Bone proteins PHEX and DMP1 regulate fibroblastic growth factor Fgf23 expression in osteocytes through a common pathway involving FGF receptor (FGFR) signaling.

Fibroblastic growth factor 23 (FGF23) is a circulating phosphaturic hormone. Inactivating mutations of the endopeptidase PHEX or the SIBLING protein DMP1 result in equivalent intrinsic bone mineralization defects and increased Fgf23 expression in osteocytes. The mechanisms whereby PHEX and DMP1 regulate Fgf23 expression are unknown. We examined the possibility that PHEX and DMP1 regulate Fgf23 through a common pathway by analyzing the phenotype of compound Phex and Dmp1 mutant mice (Hyp/Dmp1(-/-)). Compared to single-mutant littermates, compound-mutant Hyp/Dmp1(-/-) mice displayed nonadditive elevations of serum FGF23 (1912 ± 183, 1715 ± 178, and 1799 ± 181 pg/ml), hypophosphatemia (P(i): 6.0 ± 0.3, 5.8 ± 0.2, and 5.4 ± 0.1 mg/dl), and severity of rickets/osteomalacia (bone mineral density: -36, -36, and -30%). Microarray analysis of long bones identified gene expression profiles implicating common activation of the FGFR pathway in all the mutant groups. Furthermore, inhibiting FGFR signaling using SU5402 in Hyp- and Dmp1(-/-)-derived bone marrow stromal cells prevented the increase in Fgf23 mRNA expression (129- and 124-fold increase in Hyp and Dmp1(-/-) vs. 1.3-fold in Hyp+SU5402 and 2.5-fold in Dmp1(-/-)+SU5402, P<0.05). For all analyses, samples collected from nonmutant wild-type littermates served as controls. These findings indicate that PHEX and DMP1 control a common pathway regulating bone mineralization and FGF23 production, the latter involving activation of the FGFR signaling in osteocytes.

Mislocalized scaffolding by the Na-H exchanger NHE1 dominantly inhibits fibronectin production and TGF-beta activation.

Secretion and assembly of the extracellular matrix protein fibronectin regulates a number of normal cell and tissue functions and is dysregulated in disease states such as fibrosis, diabetes, and cancer. We found that mislocalized scaffolding by the plasma membrane Na-H exchanger NHE1 suppresses fibronectin expression, secretion, and assembly. In fibroblasts, wild-type NHE1 localizes to the distal margin of membrane protrusions or lamellipodia but a mutant NHE1-KRA2 lacking binding sites for PI(4,5)P2 and the ERM proteins ezrin, radixin, and moesin is mislocalized and found uniformly along the plasma membrane. Although NHE1 regulates intracellular pH homeostasis, fibronectin production is not regulated by changes in intracellular pH, nor is it attenuated in NHE1-deficient cells, indicating fibronectin expression is independent of NHE1 activity. However, fibronectin production is nearly absent in cells expressing NHE1-KRA2 because scaffolding by NHE1 is mislocalized. Additionally, secretion of active but not latent TGF-beta is reduced and exogenous TGF-beta restores fibronectin secretion and assembly. Our data indicate that scaffolding by NHE1-KRA2 dominantly suppresses fibronectin synthesis and TGF-beta activation, and they suggest that NHE1-KRA2 can be used for obtaining a mechanistic understanding of how fibronectin production is regulated and speculatively for therapeutic control of dysregulated production in pathological conditions.

Positive feedback between Cdc42 activity and H+ efflux by the Na-H exchanger NHE1 for polarity of migrating cells.

A fundamental feature of cell polarity in response to spatial cues is asymmetric amplification of molecules generated by positive feedback signaling. We report a positive feedback loop between the guanosine triphosphatase Cdc42, a central determinant in eukaryotic cell polarity, and H(+) efflux by Na-H(+) exchanger 1 (NHE1), which is necessary at the front of migrating cells for polarity and directional motility. In response to migratory cues, Cdc42 is not activated in fibroblasts expressing a mutant NHE1 that lacks H(+) efflux, and wild-type NHE1 is not activated in fibroblasts expressing mutationally inactive Cdc42-N17. H(+) efflux by NHE1 is not necessary for release of Cdc42-guanosine diphosphate (GDP) from Rho GDP dissociation inhibitor or for the membrane recruitment of Cdc42 but is required for GTP binding by Cdc42 catalyzed by a guanine nucleotide exchange factor (GEF). Data indicate that GEF binding to phosphotidylinositol 4,5-bisphosphate is pH dependent, suggesting a mechanism for how H(+) efflux by NHE1 promotes Cdc42 activity to generate a positive feedback signal necessary for polarity in migrating cells.