Application of Medical Imaging and 3D Printing Technology in Teaching the Handling of Novel Medicine in Periodontal Surgery.

Recently, fibroblast growth factor-2 (FGF-2) agents for periodontal tissue regeneration have been increasingly applied to the treatment of periodontal disease. Our current challenge for resident dentists with little clinical experience is to enhance instruction in the handling of new medicine in addition to teaching conventional procedures in periodontal tissue regeneration. This report describes using case-specific, cost-effective three-dimensional (3D) models for dentists' lectures and periodontal surgical training. As an educational and training aid, preoperative and postoperative cone-beam computed tomography images were superimposed to enable three-dimensional observation of postoperative bone regeneration. A three-dimensional anatomical model was fabricated based on these images. Dental laboratory materials were used to reproduce the periosteum and gum. The fabrication time per 3D model was about 2 hours and the cost per model was about $0.5. These models were used for lectures to resident dentists and periodontal surgery training, and their feedback was obtained. The resident's response to surgical training using these 3D models was generally positive. The use of FGF-2 represents a new direction in the treatment of periodontal disease. This being new, however, means that inexperienced periodontists require training in its application and how this will affect prognosis, as this will differ from that with more conventional techniques aimed at tissue regeneration. The low-cost 3D model presented in this report can be a valuable tool to help accomplish this in teaching inexperienced dentists, such as resident dentists.

Single shot 3D profilometry by polarization pattern projection.

We demonstrate a uniaxial 3D profilometry system illuminating the sample with a linear polarization pattern and measuring a polarization camera. The linear polarization pattern is generated by a spatial light modulator and a quarter-wave plate in the optical system. The system can measure four different fringe patterns with a phase difference of 90 deg simultaneously in the polarization camera. Therefore, we can measure three-dimensional shapes in a single shot. We present the measurement principles of the system and show the results of a real-time 3D profilometry experiment.

Temporal and dynamic changes in gingival blood flow during progression of ligature-induced periodontitis.

OBJECTIVES: To evaluate temporal changes in gingival blood flow (GBF) during progression of periodontitis in rats using a laser Doppler flowmeter (LDF) approach and to characterize morphological and biochemical features in the periodontium associated with GBF. MATERIALS AND METHODS: Forty-two Wistar rats were divided into a ligature-induced periodontitis group and a control group. To induce periodontitis, ligatures were tied around maxillary first molars bilaterally. GBF was measured in palatal gingiva at pretreatment and following ligature placement after 30 min, 1, 3, 7, 14, 21, and 28 days using LDF with a non-contact probe. Bone loss and gene expression in gingival tissues were assessed using micro-computed tomography (µCT) and quantitative polymerase chain reaction (PCR), respectively. Immunostaining for vascular endothelial growth factor (VEGF) in the maxilla was also histologically evaluated. RESULTS: GBF in the ligature group increased significantly compared with the control group 30 min after ligation. However, on days 3 and 7, GBF decreased in the ligature group. Also, after day 10, there was no difference in GBF between groups. The levels of alveolar bone loss, gene expression (interleukin-6, cluster of differentiation-31, VEGF-A, and lymphatic vessel endothelial hyaluronan receptor-1), and immunostained VEGF-positive vessels correlated well with changes in GBF. CONCLUSION PROGRESSION OF PERIODONTITIS: In rats was associated with a triphasic pattern of GBF, consisting of a short initial increase, followed by a rapid decrease, and then a gradual plateau phase.

Distribution of glutamate receptor, ionotropic, kainate 1 and neuropeptide calcitonin gene-related peptide mRNAs during formation of the embryonic and postnatal mouse molar in the maxilla.

The neuropeptide calcitonin gene-related peptide (CGRP) is a well-characterized neurotransmitter. Glutamate receptor, ionotropic, kainate 1 (Grik1) has also been demonstrated to generate high-affinity kainate receptors. However, little is known about the roles of CGRP and Grik1 during the developmental formation of teeth. In this study, we endeavoured to analyse the expression and localization of CGRP and Grik1 mRNAs using in situ hybridization on the mouse maxilla during development from the embryonic stage (E18.5) to after birth (P10, P15 and P20). We found that hybridization with an anti-sense probe for CGRP clearly localized in the maxilla at E18.5 in contrast to that of P15 and P20. Hybridization with an anti-sense probe for CGRP was not detected in the dental pulp of molars in the maxilla at P10, which is in contrast to Grik1 mRNA at the same developmental stage. Hybridization with an anti-sense probe for Grik1 mRNA was detected in the basal region of the dental pulp of molars at P10 and P15. Finally, these markers were not detected in molars in the mouse maxilla at P20. The ratio of positive cells for the hybridization signals of Grik1and CGRP in the dental pulp decreased from E18.5 (p<0.001). These features in CGRP and Grik1r mRNAs may indicate roles of function during tooth development between embryonic and postnatal stages with root formation and erupted movements.

Distribution of the neuropeptide calcitonin gene-related peptide-α of tooth germ during formation of the mouse mandible.

Calcitonin gene-related peptide-α (CGRPα) is a neurotransmitter that is related to bone formation during development. However, CGRP expression is not well known to affect the formation of teeth during development. During tooth germ development, the relationships among CGRPα, calcitonin receptor-like receptor (CRLR), amelogenin (AMELX), dentin sialophosphoprotein (DSPP), osteopontin (OPN) and osteocalcin (OCN) are unclear despite various tooth and osteogenesis markers. Our real-time RT-PCR results showed that the expression levels of CGRPα mRNA gradually decreased, in contrast to the mRNA abundances of CRLR, AMELX, DSPP, OPN, and OCN, which rapidly increased from E14.5 to P1 in the mandible. In situ hybridization using an antisense probe for CGRPα mRNA showed significant localized expression levels around the tooth bud at E14.5 and epithelial cells near the dental ledge and outer and inner enamel epithelium at E17.5 compared to those at P1. The localization of the anti-CGRPα antibody reaction revealed a strong positive reaction at the surface layer of oral epithelial cells at E14.5 and oral epithelial cells of the dental lamina around the dental ledge depression in the mandible of E17.5 mice using immunohistochemical methods The different anti-CGRPα reaction revealed its important roles during tooth formation at the postnatal stage. CGRPα mRNA was also detected in the interactions of tooth germ with the formation of odontoblast and amelobast layers from dental papilla and inner enamel epithelium. CGRPα may also be related to tooth germ development. Furthermore, CGRPα is an important tooth and bone formation marker, and bone cells provide further evidence of a role in mandibular development in contrast to inflammatory systems.

Morphological observation and CBCT of the bony canal structure of the groove and the location of blood vessels and nerves in the palatine of elderly human cadavers.

PURPOSE: The greater and lesser palatine nerves and vessels supply the hard and soft palates, and the roots of these vessels and nerves run through a bony structure. However, the arrangement of blood vessels in the maxilla requires attention during clinical treatments, but detailed morphological information about changes in the greater and lesser palatine arteries and nerves during aging is unavailable. We therefore need detailed investigations of the morphology of the donor cadaver palatine using cone-beam computed tomography (CBCT) and macroscopic observations. METHODS: We investigated 72 donor cadavers using macroscopic segmentation and CBCT. The results' analysis examined differences in skull measurement parameters and differences between dentate and edentulous cases. RESULTS: The greater palatine artery and nerve showed different macroscopic arrangements in dentate and edentulous cadavers. We also classified three types of bony structures of the nerve and vessel roots in the molar regions of the palatine using CBCT images: the shallow groove, deep groove, and flat groove. The deep groove is the deepest of the three and is remarkable in edentulous elderly cadavers. CONCLUSION: This study of macroscopic and CBCT data provides information useful for planning dental implant surgeries and autogenous bone harvesting.

Expression of CGRP, vasculogenesis and osteogenesis associated mRNAs in the developing mouse mandible and tibia.

The neuropeptide Calcitonin Gene-Related Peptide (CGRP) is a well-characterized neurotransmitter. However, little is known about the role of CGRP in osteogenesis and vascular genesis during the developmental formation of bone. In the present study, we assessed the abundance of CGRP mRNA and the mRNA of osteogenesis and vascular genesis markers in the foetal mouse mandible and leg bone (tibia). We also analysed the expression and localization of CGRP, osteopontin (OPN) and vascular endothelial growth factor (VEGF-A) using in situ hybridization and immunohistochemical localization in the mouse mandible and tibia at embryonic days 12.5 (E12.5), E14.5, E17.5, and postnatal day 1 (P1). CGRP was clearly detected in the mandible relative to the tibia at E14.5. Hybridization using an anti-sense probe for CGRP was not detected in the mandible at P1. Hybridization with an anti-sense probe for OPN was detected at E14.5, later in the mandible and at P1 in Meckel's cartilage. However, OPN was only detected in the tibia at E17.5 and later. The abundance of CGRP mRNA differed between the mandible and tibia. The level of vasculogenesis markers, such as VEGF-A, was similar to that of CGRP in the mandible. The levels of VEGF-A, cluster of differentiation 31 (CD31) and lymphatic vessel endothelial hyaluronan receptor 1 (LIVE-1) differed from that of OPN in the mandible. In contrast, the levels of VEGF-A, CD31, matrix metalloproteinase-2 (MMP-2), collagen I (Col I), collagen II (Col II) and OPN mRNA differed from E12.5 to P1 (P<0.001) in the tibia. The abundance of mRNA of CGRP and bone matrix markers (Col I, Col II, and OPN) was low at P5 in the tibia. These differences in CGRP and other mRNAs may induce a different manner of ossification between the mandible and tibia. Therefore, a time lag of ossification occurs between the mandible and tibia during foetal development.

A Simplified Method for Three-Dimensional (3-D) Ovarian Tissue Culture Yielding Oocytes Competent to Produce Full-Term Offspring in Mice.

In vitro growth of follicles is a promising technology to generate large quantities of competent oocytes from immature follicles and could expand the potential of assisted reproductive technologies (ART). Isolated follicle culture is currently the primary method used to develop and mature follicles in vitro. However, this procedure typically requires complicated, time-consuming procedures, as well as destruction of the normal ovarian microenvironment. Here we describe a simplified 3-D ovarian culture system that can be used to mature multilayered secondary follicles into antral follicles, generating developmentally competent oocytes in vitro. Ovaries recovered from mice at 14 days of age were cut into 8 pieces and placed onto a thick Matrigel drop (3-D culture) for 10 days of culture. As a control, ovarian pieces were cultured on a membrane filter without any Matrigel drop (Membrane culture). We also evaluated the effect of activin A treatment on follicle growth within the ovarian pieces with or without Matrigel support. Thus we tested four different culture conditions: C (Membrane/activin-), A (Membrane/activin+), M (Matrigel/activin-), and M+A (Matrigel/activin+). We found that the cultured follicles and oocytes steadily increased in size regardless of the culture condition used. However, antral cavity formation occurred only in the follicles grown in the 3-D culture system (M, M+A). Following ovarian tissue culture, full-grown GV oocytes were isolated from the larger follicles to evaluate their developmental competence by subjecting them to in vitro maturation (IVM) and in vitro fertilization (IVF). Maturation and fertilization rates were higher using oocytes grown in 3-D culture (M, M+A) than with those grown in membrane culture (C, A). In particular, activin A treatment further improved 3-D culture (M+A) success. Following IVF, two-cell embryos were transferred to recipients to generate full-term offspring. In summary, this simple and easy 3-D ovarian culture system using a Matrigel drop and activin A supplementation (M+A) provides optimal and convenient conditions to support growth of developmentally competent oocytes in vitro.

A new spin-polarized photoemission spectrometer with very high efficiency and energy resolution.

A new spin- and angle-resolved photoemission spectrometer was developed adopting the very-low-energy-electron-diffraction (VLEED)-type spin polarimeter. The Fe(001)p(1x1)-O film grown on MgO(001) crystal for the VLEED target yields significantly high spin-resolving power, the effective Sherman function of 0.40+/-0.02, with long lifetime and stability compared to the conventional Fe(001) target. Under the favor of high resolving power, approximately 100 times higher efficiency than that of conventional Mott-type spin polarimeter, the figure of merit of 1.9+/-0.2x10(-2) was achieved. Owing to this high efficiency, high-energy resolution can be realized with this new spin-polarized photoemission spectrometer. The simplified ways of target preparation and revitalization make the VLEED spin polarimeter much more convenient and feasible for the spin-polarized photoemission spectroscopy.