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excess sugar, poor oral hygiene) or consequences (eg, pain, appearance). Knowledge did not vary by men’s socio-demographic characteristics or acculturation level. Few men recognized early stages of the disease, with most fathers describing the appearance of caries in two major ways: by the structure of the tooth (eg, crumbling) and how it looked (eg, black, holes.) Conclusion: No matter how long men had been in the US, Mexican-origin fathers displayed limited knowledge of caries as a disease process when compared to the definition used by most health professionals. They also described advanced stages of caries, which suggests late recognition of the caries process, possibly leading to delay in seeking care for their children’s oral problems. Support: The UCSF Program in Global Oral Health, NIH/NIDCR grant # R21 DE019210; #06 Title: Notch signaling is required for attachment of ameloblasts to the stratum intermedium R ESPINOZA (1), A H JHEON (2), Y LIM (3), T WEN (2), B BIEHS (2), and O D KLEIN (2, 4)* (1) UCSF School of Dentistry (2) Department of Orofacial Sciences and Program in Craniofacial and Mesenchymal Biology, UCSF (3) Seoul National University, Korea (4) Department of Pediatrics and Institute for Human Genetics, UCSF *Corresponding author: ophir.klein@ucsf.edu Introduction: Little is known about the roles of Notch signaling and cell-cell adhesion during tooth development. Here we utilize the continuously growing rodent incisor to determine that Notch signaling is an important regulator of enamel formation through effects on the interaction between enamel-forming ameloblasts and its subjacent attachment layer, the stratum intermedium (SI). Objective: To examine the role of Notch signaling during amelogenesis in vivo and in vitro. Methods: Inhibitory antibodies specific for components of the Notch signaling pathway (Notch1, Notch2, Jag1, Jag2) were injected into the peritoneum of adult mice for two weeks. The lower incisors were then excised, sectioned, and evaluated using H&E staining and immunohistochemistry. Notch signaling was inhibited in the ameloblast-like LS8 cell line and the relative expression levels of genes downstream of Notch signaling were assayed using real-time quantitative PCR. Results: The inhibition of the Notch signaling pathway resulted in detachment between the ameloblasts and the SI. Furthermore, there were changes in the localization of PERP and desmoplakin but not amelogenin and ameloblastin. PERP is a transmembrane protein that is required in the formation and homeostasis of desmosomes, which are specialized cell structures for cell-cell adhesion. Desmoplakin is a specific component of desmosomes. Previously, a similar detachment of the ameloblast-SI interface was observed in Perp-null mice. Inhibition of Notch signaling in vitro led to decreases in Perp expression, demonstrating that Notch signaling is involved in the regulation of Perp. Conclusion: There is little prior knowledge regarding the role of Notch signaling during tooth development. Here we show for the first time that Notch signaling is required for the integrity of the ameloblast-SI interface through its regulation of Perp, a gene important in cell-cell adhesion. Support: NIH R21-DE019958 #07 Title: The role of peptidoglycan recognition protein in Fusobacterium nucleatum induced apoptosis in Jurkat T cells K EVANS (1), M RAPOZO-HILO (2), Y RAVANDOUST (2), D FRASER (1), C HOOVER (3), and T HUYNH (2) (1) UCSF School of Dentistry (2) Department of Preventive and Restorative Dental Sciences, UCSF School of Dentistry (3) Department of Cell & Tissue Biology, Department of Microbiology & Immunology, UCSF Objectives: Fusobacterium nucleatum (Fn), a gram-negative oral anaerobe, has been implicated in the pathogenesis of infections throughout the body including periodontal and endodontic infections. Fn’s role in these infections remains unknown. We have shown that Fn can induce apoptosis in immune cells (Jewett 2000). The mechanism of this phenomenon remains unclear but it appears that aggregation of immune cells and upregulation of death ligands play a significant role (Huynh 2011, Fraser 2010). Peptidoglycan recognition proteins (PGRP-S) are responsible for detecting bacteria-derived peptidoglycans. Recently, it was shown that lymphocytic PGRP-S participated in target recognition and induced death ligand mediated cell apoptosis (Sashchenko 2007). The purpose of this study was to determine if PGRP-S has a direct role in Fn induced apoptosis in Jurkat T cells. Methods: After Jurkat T cells were co-cultured with Fn for four hours, we assessed the expression of PGRP-S on Jurkat T cells via flow cytometry (MoFlo Systems). After co-culturing Fn with Jurkat T cells overnight with neutralizing antibodies to PGRP-S, the levels of Jurkat T cell apoptosis was assessed via caspase assay according to manufacturer instructions (Promega Caspase-Glo 3/7 Assay Kit). ANOVA with post-hoc Tukey’s test was used for statistical analysis at p
#08 Title: Biomechanics of a bone-tooth fibrous joint J GREENE (2), H OZCOBAN (1), D CURTIS (2), M RYDER (3), S HO (2) (1) Institute of Advanced Ceramics, Hamburg University of Technology; (2) UCSF Department of Preventive and Restorative Dental Sciences, (3) UCSF Department of Orofacial Sciences Objective: To investigate biomechanical responses of healthy and diseased bone-tooth fibrous joints to mechanical loads. Methods: Periodontal disease was induced using a ligature around the 2nd molar of four month old male rats. Control specimens were flossed every 4 days to ensure mechanical distress due to ligature itself. After four weeks, the right maxillae were harvested from control (n=6) and diseased (n=6) specimens, and only the second molar was loaded. All specimens were compressed at 10µm per second; to a maximum displacement of 70µm. Results: Load-displacement curve of the bone-tooth fibrous joint consisted of two segments: 1) a nonlinear segment of lower stiffness (S1) followed by 2) a linear stiffer segment (S2). The first segment (displacement from 0-32μm) was used to evaluate an equivalent stiffness of PDL, while the second segment (displacement from 32-70μm) was more representative of compression related effects of the bone-tooth complex. Instrument sensitivity was validated by compressing chemically-fixed diseased specimens. S1 results for control, diseased and diseased-fixed specimens were: 45±25, 30±17, 134±16 N/mm respectively. S2 results for control, diseased and diseased-fixed specimens were: 91±53, 81±42, 214±22 N/mm respectively. A significant difference in stiffness was observed between diseased and diseased-fixed specimens (p0.05) using Student& rsquo;s t-test with a 95% confidence interval. Conclusions: Although results were not significant, trends indicate diseased specimens had decreased PDL stiffness and damping viscoelasticity as shown by S1 and less bone compression as shown by S2. Additional studies evaluated over longer periods would provide further information on how periodontal disease affects biomechanics of dentition. Support: UCSF Department of Preventive and Restorative Dental Sciences,UCSF Department of Orofacial Sciences, NIH R00DE018212, S10RR026645 #09 Title: Three-dimensional Analysis of the Morphologic Soft Tissue Changes Following Facial Reconstructive Surgery C GUERSOLI (1) L LEE (2), J HONG (3), S BEKMEZIAN (2), J HUANG (4), J LEE (2) (1) UCSF School of Dentistry (2) UCSF Department of OMFS (3) Samsung University and Medical Center, Department of OMFS, Korea (4) UCSF OFS Department, Division of Orthodontics Introduction: Soft tissue changes after orthognathic surgery have previously been recorded using 2-dimensional lateral cephalograms. This limits the analysis to landmarks measured in the midline (i.e. subnasale) and involves imaging limitations such as radiographic magnification variations. However, orthognathic advancement procedures change soft tissue in 3-dimensional areas beyond the midline and dramatically change a person’s appearance in these regions. Objectives: This study determined the 3-dimensional effects of maxillary advancement through quantification of the changes in the perinasal region and correlation of these morphologic changes to maxillary skeletal change. Methods: Patients undergoing maxillary advancement surgery were enrolled in this CHR-approved study. We used 3-dimensional cone-beam computed-tomography (CBCT) images in conjunction with InVivo5 software (AnatomageTM, San Jose, CA) to measure soft tissue changes at the traditional midsagittal landmarks (pronasale, soft tissue A-point, upper and lower lip anterior points) as well as points extending horizontally to the right and left 10-mm, 25-mm, and 35-mm from the three superior midsagittal landmarks. The immediate pre-operative and six-month follow-up CBCTs of 30 patients were analyzed for this study. Results: Soft tissue change extended into the infraorbital region from the lower lip, and laterally to the zygomatic region, well beyond the midline structures. The areas of strongest correlation were at 10-mm left of the soft-tissue A-point, and at midline and 10-mm left of the upper lip anterior point (Pearson correlation=0.909, 0.897, and 0.848 respectively, p
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