Naoko Kubota

Jaw motion during gum-chewing in children with primary dentition.

Abstract This study was undertaken to characterize jaw motion during mastication in children with primary dentition and to compare jaw motion with that in adults. The means and the variances of the traditional parameters for the chewing cycle, i.e., duration, excursive ranges and 3-D distances of travel at the lower incisor, molars and condyles were analyzed and compared in 23 children and 25 female adults. The duration of opening in children was significantly shorter than that of adults. Significant differences between children and adults were observed in lateral and vertical excursion of the incisor, lateral excursion at the molars, and vertical excursion at the condyles. Many of these measurements had larger between-subject and between-cycle variances in children than adults, suggesting that chewing motion in children has not yet matured. The results of this study indicate that chewing motion in children is different from that of adults.

A reproducibility method to test lip-closing strength in preschool children.

Abstract This cross-sectional study tested the reproducibility of a simple button-pulling system for measuring lip-closing strength in normal preschool children and compared their strength to that of normal adults. The sample consisted of 348 preschool children and 123 adults. Lip-closing strength was measured by inserting a button, fastened to a piece of string, into the vestibule between the incisors and lips with minimal mouth opening. The string was attached to a digital tension gauge and was pulled parallel to the floor. Maximum tension, with three repetitions, was recorded at the instant that the button was pulled out of the mouth. Multilevel statistical models were used to evaluate any differences in contractive muscle strength between age groups and between the genders. The strength in children increased significantly from three years to five years (p<0.01). Gender-related differences were found in adults but not in preschool children. Inter-individual variation at each age was larger than intra-individual variation. Measurement of lip-closing strength by button pulling is highly reproducible in children and has potential clinical and research applications.

Choice of Feeders Is Important When First Establishing iPSCs Derived From Primarily Cultured Human Deciduous Tooth Dental Pulp Cells

Feeder cells are generally required to maintain embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs). Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used feeder cells. The aim of this study was to determine which cells are suitable for establishing iPSCs from human deciduous tooth dental pulp cells (HDDPCs). Primary cultures of HDDPCs were cotransfected with three plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 and pmaxGFP by using a novel electroporation method, and then cultured in an ESC qualified medium for 15 days. Emerging colonies were reseeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether cells exhibited ESC-like morphology and alkaline phosphatase activity to evaluate the state of cellular reprogramming. HDDPCs maintained on MEFs were successfully reprogrammed into iPSCs, whereas those maintained on STO cells were not. Once established, the iPSCs were maintained on STO cells without loss of pluripotency. Our results indicate that MEFs are better feeder cells than STO cells for establishing iPSCs. Feeder choice is a key factor enabling efficient generation of iPSCs.

Alkaline phosphatase and OCT-3/4 as useful markers for predicting susceptibility of human deciduous teeth-derived dental pulp cells to reprogramming factor-induced iPS cells

AIM The aim of the present study was to prove that primary cells enriched with stem cells are more easily reprogrammed to generate induced pluripotent stem (iPS) cells than those with scarce numbers of stem cells. METHODS We surveyed the alkaline phosphatase (ALP) activity in five primarily-isolated human deciduous teeth-derived dental pulp cells (HDDPC) with cytochemical staining to examine the possible presence of stem cells. Next, the expression of stemness-specific factors, such as OCT(Octumer-binding transcription factor)3/4, NANOG, SOX2(SRY (sex determining region Y)-box 2), CD90, muscle segment homeodomain homeobox (MSX) 1, and MSX2, was assessed with a reverse transcription polymerase chain reaction method. Finally, these isolated HDDPC were transfected with plasmids carrying genes coding Yamanaka factors to determine whether these cells could be reprogrammed to generate iPS cells. RESULTS Of the five primarily-isolated HDDPC, two (HDDPC-1 and -5) exhibited higher degrees of ALP activity. OCT-3/4 expression was also prominent in those two lines. Furthermore, these two lines proliferated faster than the other three lines. The transfection of HDDPC with Yamanaka factors resulted in the generation of iPS cells from HDDPC-1 and -5. CONCLUSION The number of cells with the stemness property of HDDPC differs among individuals, which suggests that HDDPC showing an increased expression of both ALP and OCT-3/4 can be more easily reprogrammed to generate iPS cells after the forced expression of reprogramming factors.

Relationship between nasal and skeletal landmarks on lateral cephalograms of adults

The purpose of this study was to investigate the relationship between cephalometric nasal soft tissue and skeletal landmarks in adults. Lateral cephalograms from Japanese adults (30 men: mean age, 24.5 ± 4.9 years; 30 women: mean age, 20.3 ± 3.3 years; overall mean age, 22.4 ± 2.4 years) were used in this study. Twenty-two skeletal points and three soft tissue nasal points were marked on each subject’s lateral cephalogram, and the coordinates of all the points were systematically digitised and transformed to a standardised plane. A forward stepwise regression analysis determined how combinations of the skeletal landmarks predict the location of the nasal soft tissue landmarks. Based on the result of our research, the location of the nasal soft tissue cephalometric landmarks in our adult subjects may be predicted based on skeletal landmarks, and different skeletal landmarks predicted the position of each soft tissue landmark in the adult males and females in this study.

Hela Cells Consist of Two Cell Types, as Evidenced by Cytochemical Staining for Alkaline Phosphatase Activity: A Possible Model for Cancer Stem Cell Study

Alkaline phosphatase (ALP) is known to be expressed in the several somatic stem cells and cancer cells. To investigate whether ALP may be a promising marker for cancer stem cells (CSCs), we examined the expression of ALP in human squamous cell carcinoma HeLa cells using a cytochemical

Comparison of normal permanent and primary dentition sagittal tooth-crown inclinations of Japanese females.

Abstract The purpose of this study was to clarify the characteristics of permanent and primary toothcrown inclinations. Landmark points from cephalograms and dental casts of two groups; 23 women (mean 20.3±3.3 years) and 11 girls (mean 5.2±0.1 years) were digitized, and the coordinates were integrated and transformed to a standardized plane. The 3-dimensional crown inclinations were projected on the sagittal plane, and the angles between the tooth vectors and the FH plane were calculated. An independent-group t-test was used to test for group differences of each tooth inclination, and correlation coefficients were generated for the inclination angles among the permanent and primary teeth. Most maxillary tooth-crown inclinations showed significant age-related differences, while only the second premolar and primary second molar differed significantly in the mandible. The maxillary molars were parallel to the corresponding mandibular molars and correlated with each other, but the primary molars were not. Significant correlations were found between inclinations of most permanent teeth, but not the primary teeth. Maxillary tooth-crown inclinations change during growth, but tooth-crown inclinations of the mandibular teeth do not.

Intrapancreatic Parenchymal Injection of Cells as a Useful Tool for Allowing a Small Number of Proliferative Cells to Grow In Vivo

In vivo inoculation of cells such as tumor cells and induced pluripotent stem (iPS)/embryonic stem (ES) cells into immunocompromised mice has been considered as a powerful technique to evaluate their potential to proliferate or differentiate into various cell types originating from three germ cell layers. Subcutaneous grafting and grafting under the kidney capsule have been widely used for this purpose, but there are some demerits such as the requirement of a large number of tumor cells for inoculation and frequent failure of tumorigenesis. Therefore, grafting into other sites has been explored, including intratesticular or intramuscular grafting as well as grafting into the cochleae, liver, or salivary glands. In this study, we found that intrapancreatic parenchymal injection of cells is useful for allowing a small number of cells (~15 × 103 cells or ~30 cell clumps μL−1·site−1) to proliferate and sometimes differentiate into various types of cells. It requires only surgical exposure of the pancreas over the dorsal skin and subsequent injection of cells towards the pancreatic parenchyma under dissecting microscope-based observation using a mouthpiece-controlled glass micropipette. We now name this technology “intrapancreatic parenchymal cell transplantation (IPPCT)”, which will be useful, especially when only a small number of cells or colonies are available.

Effect of Storage Conditions on Biaxial Flexure Strengths of MTA

The aim of this study was to determine the biaxial flexure strength of a commercially available dental pulp capping or cavity liner material, mineral trioxide aggregate (MTA). MTA and a conventional pulp capping material, calcium hydroxide, were used in this study. Each material was prepared into a disk-shaped specimen. Those specimens were stored in 100% humidity atmosphere at 23 or 37°C for 3 or 24 h. Six specimens in each condition were used for the biaxial flexure test using a piston-on-three-ball according to ISO-6872. Surface pH values were determined. The biaxial flexure strengths of MTA after setting in 24h were larger than those of Dycal (p <0.01). pH value of MTA was higher than that of Dycal. These results indicated that MTA was clinically usage material.