Effect of fibroblast growth factor and enamel matrix derivative treatment on root resorption after delayed replantation.

BACKGROUND: Periodontal ligament (PDL) healing and long term prognosis of replanted avulsed teeth should rely on several factors including length of extra-oral dry time and type of the storage medium. The status of periodontal ligament is critical for the healing of replanted teeth. Different substances have been used for root surface treatment to promote formation of PDL and increase the survival of avulsed teeth submitted to replantation. AIM: The purpose of this study was to assess the effect of recombinant basic fibroblast growth factor 2 (bFGF) and enamel matrix derivative (EMD) on root resorption after delayed replantation. DESIGN: 18 freshly extracted single-rooted incisor and premolar teeth were extracted from the beagle dogs and immersed in whole bovine milk for 45 and 60 min (n = 3 each). Following storage period, sockets washed and teeth were treated with bFGF and EMD and replanted into the sockets. After 8 weeks, dogs were sacrificed, specimens processed to 4-µm thick serial sections for histopathologic examination and morphometric assessments. Thus, the proportions of the roots that exhibited signs of surface resorption, inflammatory resorption, and replacement resorption, that is, ankylosis and normal PDL were noted. RESULTS: The percentage of root resorption was in the following order: EMD>milk>bFGF for 45 min and milk>EMD>bFGF for 60 min. For all groups, teeth stored 60 min showed significantly higher incidence of PDL resorption than those stored for 45 min (P < 0.01). The highest incidence of replacement resorption was observed in teeth treated with EMD for 60 min. After 8 weeks, the least resorption was found in bFGF-treated group (P < 0.01). CONCLUSIONS: The findings of this study suggest that use of bFGF favored the formation of new periodontal ligament; prevent ankylosis and resorption process following delayed replantation of teeth while EMD shows replacement resorption, which may turn to ankylosis.

Genetic mapping of agenesis of the third molars in mice.

EL/Sea mice are characterized by 100% incidence of agenesis of the third molars (M3). In a previous study, chromosomal mapping of the ninth generation EL/Sea congenic strains revealed a major locus for agenesis of M3, designated am3, in the 125-137 Mbp region of chromosome 3. In the present study, to determine the precise location of the am3 locus, we produced further generations of the EL/Sea congenic strains for am3 in which the restricted interval on chromosome 3 of EL/Sea was replaced by a MSM/Msf-derived homolog. The eleventh generation congenic mice that were either heterozygous or homozygous for the MSM/Msf-derived interval exhibited a significant decrease in the incidence of M3 agenesis (p < 0.00001). Results confined the am3 locus to an interval of 1 Mbp on chromosome 3, demonstrating that Lef1, one of the essential transcription factors for early tooth development, is the strongest candidate for am3.

Mutations of the SH3BP2 gene in 2 families of cherubism.

PURPOSE: Cherubism is a rare autosomal dominant syndrome characterized by abnormal bone tissue in the lower part of the face. Mutations in the gene coding for SH3BP2 have been identified in about 80% of people with cherubism. The aim of this study was to determine whether a mutation in the SH3BP2 gene was the molecular basis of cherubism in two unrelated families. METHODS: Two cases of the aggressive form of Cherubism were described in two Turkish families with extensive bilateral swelling in the mandible, typical pathological features and familial history. Genomic DNA was extracted from six affected and three unaffected individuals from two families, and mutations in the SH3BP2 were detected by PCR, and direct DNA sequencing was carried out. RESULTS: In the first family, a missense mutation Arg415Gln was found in exon 9 of the SH3BP2 in all affected individuals. The unaffected individuals did not have this mutation. In the second family, another missense mutation Pro418Thr was identified in exon 9 of the SH3BP2 in the patient and his mother with cherubism. CONCLUSIONS: We detected the point mutations in the SH3BP2 gene in the patients with multiple affected individuals. Genotype-phenotype association studies in individuals with cherubism are necessary to provide important knowledge about molecular mechanisms of the disease.

Roles of salivary components in Streptococcus mutans colonization in a new animal model using NOD/SCID.e2f1-/- mice.

Streptococcus mutans plays an important role in biofilm formation on the tooth surface and is the primary causative agent of dental caries. The binding of S. mutans to the salivary pellicle is of considerable etiologic significance and is important in biofilm development. Recently, we produced NOD/SCID.e2f1(-/-) mice that show hyposalivation, lower salivary antibody, and an extended life span compared to the parent strain: NOD.e2f1(-/-). In this study we used NOD/SCID.e2f1(-/-) 4 or 6 mice to determine the roles of several salivary components in S. mutans colonization in vivo. S. mutans colonization in NOD/SCID.e2f1(-/-) mice was significantly increased when mice were pre-treated with human saliva or commercial salivary components. Interestingly, pre-treatment with secretory IgA (sIgA) at physiological concentrations promoted significant colonization of S. mutans compared with sIgA at higher concentrations, or with human saliva or other components. Our data suggest the principal effects of specific sIgA on S. mutans occur during S. mutans colonization, where the appropriate concentration of specific sIgA may serve as an anti-microbial agent, agglutinin, or an adherence receptor to surface antigens. Further, specific sIgA supported biofilm formation when the mice were supplied 1% sucrose water and a non-sucrose diet. The data suggests that there are multiple effects exerted by sIgA in S. mutans colonization, with synergistic effects evident under the condition of sIgA and limited nutrients on colonization in NOD/SCID.e2f1(-/-) mice. This is a new animal model that can be used to assess prevention methods for dental biofilm-dependent diseases such as dental caries.

Craniofacial and dental characteristics of Goldenhar syndrome: a report of two cases.

We describe the dental and craniofacial anomalies of 2 ethnically distinct patients with Goldenhar syndrome, which is characterized by hemifacial microsomia, facial asymmetry, and ear and dental abnormalities. A 7-year-old Japanese girl and 12-year-old Turkish boy with Goldenhar syndrome were examined clinically and radiographically; both had symptoms of hemifacial microsomia. Multiple organ involvement can limit surgical correction of deformities and affect patient management. Therefore, long-term regular follow-up by a multidisciplinary team is important to monitor the growth and development of patients.

Prolonged survival and phenotypic correction of Akp2(-/-) hypophosphatasia mice by lentiviral gene therapy.

Hypophosphatasia (HPP) is an inherited systemic skeletal disease caused by mutations in the gene encoding the tissue-nonspecific alkaline phosphatase (TNALP) isozyme. The clinical severity of HPP varies widely, with symptoms including rickets and osteomalacia. TNALP knockout (Akp2(-/-)) mice phenotypically mimic the severe infantile form of HPP; that is, TNALP-deficient mice are born with a normal appearance but die by 20 days of age owing to growth failure, hypomineralization, and epileptic seizures. In this study, a lentiviral vector expressing a bone-targeted form of TNALP was injected into the jugular vein of newborn Akp2(-/-) mice. We found that alkaline phosphatase activity in the plasma of treated Akp2(-/-) mice increased and remained at high levels throughout the life of the animals. The treated Akp2(-/-) mice survived for more than 10 months and demonstrated normal physical activity and a healthy appearance. Epileptic seizures were completely inhibited in the treated Akp2(-/-) mice, and X-ray examination of the skeleton showed that mineralization was significantly improved by the gene therapy. These results show that severe infantile HPP in TNALP knockout mice can be treated with a single injection of lentiviral vector during the neonatal period.

Collection and culture of alveolar bone marrow multipotent mesenchymal stromal cells from older individuals.

In this work, we examined the culture condition of alveolar bone marrow multipotent mesenchymal stromal cells (ABMMSCs), aiming to apply regenerative therapy to older periodontitis patients. To better understand the character of cultured cells from alveolar bone marrow, the expression profiles of well-known genes and their responses to the induction of osteogenic, chondrogenic, or adipogenic differentiation were examined. Using alpha MEM-based culture, ABMMSCs could be obtained from older individuals than in previous reports. Interestingly, ABMMSCs expressing Klf4 were able to differentiate into osteoblasts. The prediction of differentiation potential by Klf4 could be a useful guide for further improvement of the culture conditions required to culture ABMMSCs derived from older individuals.

An orthopantomographic study of hypodontia in permanent teeth of Japanese pediatric patients.

Hypodontia of permanent teeth was evaluated from orthopantomograms of 2072 apparently healthy pediatric patients at The Hospital of Nihon University School of Dentistry at Matsudo. The prevalence of congenitally missing teeth (CMT) was 8.7% in boys and 10.8% in girls, and 9.4% for both sexes combined. Most cases (67.8%) involved either one or two missing teeth. There were in total 574 CMT, and on average 2.8 teeth were missing per child. The most commonly absent tooth was the mandibular second premolar. On the other hand, no first molars were missing in any case. A high frequency of CMT mandibular incisors (18.82%) was observed, and this seems to be a characteristic peculiar to individuals of Asian ethnicity. Oligodontia (6 or more CMT excluding the third molar) ranged from 6 to 14 teeth, with a prevalence of 1.4% in general: 1.8% for girls and 0.9% for boys. Symmetry of CMT was predominant: 214 pairs for bilateral symmetry and 107 pairs for symmetry between two antagonistic quadrants. The distribution of CMT between maxillary and mandibular hypodontia in the right and left quadrants for boys and girls no had significant association (P < 0.05).

Supernumerary tooth in the primary molar region: a case report.

Supernumerary teeth are among the most common dental anomalies affecting the primary and permanent dentition. They are usually found in the anterior maxilla and occur infrequently in the primary dentition. The purpose of this paper was to report a case diagnosed with primary supernumerary tooth in the primary second molar region. The crown and root shape of the primary supernumerary tooth resembled that of the primary first molar. On radiographic examination, the primary supernumerary tooth was followed by a permanent supernumerary successor with an unusually big crown. The primary supernumerary molar was immediately extracted to avoid interference with the development of the second premolar's tooth bud. This dental anomaly is rarely observed, as only primary supernumerary teeth in the anterior region have been reported in the dental literature.

Taste after reduction of the tongue in Beckwith-Wiedemann syndrome.

We tested the sensitivity of taste after reduction of the tongue in four girls with Beckwith-Wiedemann syndrome. No patient had taste blindness, but the ability to detect salty and bitter tastes declined after reduction of the tongue.

Enamelin (Enam) is essential for amelogenesis: ENU-induced mouse mutants as models for different clinical subtypes of human amelogenesis imperfecta (AI).

Amelogenesis imperfecta (AI) is a group of commonly inherited defects of dental enamel formation, which exhibits marked genetic and clinical heterogeneity. The genetic basis of this heterogeneity is still poorly understood. Enamelin, the affected gene product in one form of AI (AIH2), is an extracellular matrix protein that is one of the components of enamel. We isolated three ENU-induced dominant mouse mutations, M100395, M100514 and M100521, which caused AI-like phenotypes in the incisors and molars of the affected individuals. Linkage analyses mapped each of the three mutations to a region of chromosome 5 that contained the genes encoding enamelin (Enam) and ameloblastin (Ambn). Sequence analysis revealed that each mutation was a single-base substitution in Enam. M100395 (Enam(Rgsc395)) and M100514 (Enam(Rgsc514)) were putative missense mutations that caused S to I and E to G substitutions at positions 55 and 57 of the translated protein, respectively. Enam(Rgsc395) and Enam(Rgsc514) heterozygotes showed severe breakage of the enamel surface, a phenotype that resembled local hypoplastic AI. The M100521 mutation (Enam(Rgsc521)) was a T to A substitution at the splicing donor site in intron 4. This mutation resulted in a frameshift that gave rise to a premature stop codon. The transcript of the Enam(Rgsc521) mutant allele was degraded, indicating that Enam(Rgsc521) is a loss-of-function mutation. Enam(Rgsc521) heterozygotes showed a hypomaturation-type AI phenotype in the incisors, possibly due to haploinsufficiency of Enam. Enam(Rgsc521) homozygotes showed complete loss of enamel on the incisors and the molars. Thus, we report here that the Enam gene is essential for amelogenesis, and that mice with different point mutations at Enam may provide good animal models to study the different clinical subtypes of AI.

Efficacy of varying the NMEP concentrations in the NMGly-NMEP self-etching primer on the resin-tooth bonding.

It is well understood that the application of a self-etching primer enhances the bonding at the resin-teeth interface. In this study, we designed a self-etching primer consisting of N-methacryloyl glycine (NMGly) and N-methacryloyl-2-aminoethyl phosphonic acid (NMEP). The demineralization effects on the hydroxyapatite or dentin by the carboxylic acid in the NMGly and by the phosphonic acid in the NMEP and their effects on the bond strength of the resin to the tooth were examined. The application of the NMGly-NMEP solution to the enamel resulted in an increase in the bond strength when additional amounts of NMEP were added to the NMGly aqueous solution. This increase was due to the phosphonic acid in the NMEP demineralizing the enamel. Conversely, the addition of the NMEP to the NMGly solution resulted in a decrease in the bond strength to the dentin. The optimal concentration of the NMEP in the NMGly-NMEP solution resulted in bond strengths of over 20 MPa for both the enamel and dentin.

Identification of chromosomes associated with dental caries susceptibility using quantitative trait locus analysis in mice.

Dental caries is a multifactorial, infectious disease with little known about the host genetic factors influencing susceptibility. This study aimed to identify the major candidate chromosomes for dental caries susceptibility and to detect the relevant regions within these. Quantitative trait locus (QTL) analysis was performed on genetic crosses of C3H/HeJ (caries-resistant) and C57BL/6J (caries-susceptible) mice inoculated with Streptococcus mutans serotype C. In a genomewide scan, three suggestive QTLs were detected on chromosomes 1, 2, and 7, one significant QTL was found on chromosome 2, and one highly significant QTL was detected on chromosome 8. The likelihood ratio statistic (LRS) was raised around the marker D1Mit21 in the middle region of chromosome 1, between D2Mit255 and D2Mit311 in the distal region of chromosome 2, and the region distal to D7Mit31 on chromosome 7. A significant QTL was located between the markers D2Mit237 and D2Mit101 on chromosome 2. The LRS was highly significantly raised between markers D8Mit208 and D8Mit280 on chromosome 8, and exceeded a highly significant level between markers D8Mit211 and D8Mit280. These results suggest that major gene(s) responsible for dental caries susceptibility or resistance are located in one or more of these regions.