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PubMed1998

Degeneration of tooth germ in the developing dentition of the gray short-tailed opossum (Monodelphis domestica).

The aim of this study was to examine the developmental aspects of the dental lamina and the tooth germ of the marsupial opossum (Monodelphis domestica), and to clarify the dental formula of this animal. Specimens were 12-, 16-, and 18-d-old opossums. 3-D reconstructions were constructed from frontal serial sections. In these animals, the tooth germs of the deciduous maxillary and mandibular canine, deciduous third premolar and first molar, and the deciduous maxillary first incisor and second molar had a successional dental lamina and a replacement tooth germ. The tooth germ of the deciduous maxillary fourth incisor and the mandibular first incisor were reduced. The dental lamina was continuous in each jaw except for the deciduous maxillary first incisor. The first dentition (deciduous dentition) remained as the permanent dentition on the deciduous maxillary first incisor, and the deciduous maxillary and mandibular canine and first molar. The maxillary fourth incisor and the mandibular first incisor were the second dentition (successional dentition). Only the deciduous third premolars were replaced. These results showed monophyodonty caused by both deciduous and replacement tooth germ degeneration.

Animals↗
PubMed1995

[Outcome of wisdom tooth germ transplantation].

In 68 patients, 79 tooth germs of the third molar were transplanted. Out of this group, 43 patients with 50 transplantations under-went a follow-up examination. All surgery was performed by one surgeon. Retrospectively, we tried to establish whether the success of postoperative healing depends on the donor and receiver regions. Crossing the jaw border clearly worsens the prognosis for transplantation. Altogether 9 transplantations turned out to be failures, 7 of which had been transplanted from the upper to the lower jaw. The results of this investigation showed incomplete root development in 34%, the necessity of endodontic treatment in 6% and enlarged periodontal pockets in 8%. Ankylosed teeth were found in 10%; we did not see any root resorptions.

Adolescent↗
PubMed1997

Effects of actinomycin D on developing hamster molar tooth germs in vitro.

The aim of this study was to evaluate the toxic effects of actinomycin D on the developing hamster tooth germ in organ culture. Hamster tooth germs during early secretory amelogenesis were exposed in vitro for 24 h to 10(-9) M-5 x 10(-5) M actinomycin D. Actinomycin D dose-dependently (> or = 10(-7) M) decreased the tooth germ dry weight but mineralization was affected only by doses > or = 10(-5) M. However, the uptakes of TCA-insoluble 32P and [3H]thymidine were significantly reduced dose-dependently from > or = 10(-8) M actinomycin D, indicating that the drug inhibits the synthesis of phosphate-containing macromolecules as well as DNA synthesis. Histologically, 10(-8) M actinomycin D was the lowest dose which was not toxic to any cell type in the developing tooth germ. At 10(-7) M actinomycin D, the most sensitive cells were the proliferating pre-odontoblasts followed by pre-ameloblasts; the mature secretory ameloblasts and odontoblasts appeared unaffected. Higher doses resulted in increased cytotoxicity to the secretory cells and, eventually, total degeneration of most cells. The data suggest that children treated for cancer during tooth development using anti-chemotherapy cocktails containing actinomycin D (serum levels > 10(-7) M) may develop defects later on in the mature dentition as a direct consequence of the toxicity of the drug to the tooth organ.

Ameloblasts↗
PubMed1993

Reversible and irreversible effects of temperature on amelogenesis of hamster tooth germs in vitro.

Hamster first hamster molar tooth germs in early secretory stage of amelogenesis were cultured for one day in vitro at 6 degrees C, 22 degrees C, 37 degrees C or 45 degrees C in the presence of 3H-proline, 45Ca and 32P-orthophosphate. Other explants were cultured without these labels and after culture examined by histology. The highest temperature tested was lethal to the explants, decreased total dry weight and rapidly increased total uptake of the radiolabelled mineral ions, probably merely due to physicochemical modification of the existing preculture minerals. Optimal synthesis and secretion of amelogenins were measured at physiological temperature (37 degrees C). Effects of exposure to both temperatures below the physiological value were virtually reversible when explants were grown at physiological temperature (37 degrees C) for another day. However, amelogenin secretion during this recovery period did not reach values as high as those found for the first day in explants initially grown at physiological temperature during the first day. We concluded from the four temperatures examined that the optimal temperature for enamel matrix deposition in vitro was 37 degrees C. At this temperature enamel biosynthesis and its secretion are high. Lowering the temperature slows down the metabolism without any apparent harmful effect. Normal development of the tooth explants in vitro resumes when the culture temperature is restored to physiological levels (37 degrees C). For temporary storage of tooth germ explants prior to any reimplantation, we therefore recommend a temperature of 6 degrees C.

Amelogenesis↗
PubMed1980

An electron microscopic study of cell death in molar tooth germ epithelia of mouse embryos.

Cell death in the epithelial tooth germs of mouse mandibular molars from the 12th to the 14th day of gestation was investigated by electron and light microscopy. Light microscopy revealed granular substances in the epithelial portion of the tooth germ on the 12th day, and an increase in their number to the 14th day when the enamel knot developed. In the areas where granular substances were observed by the light microscope, electron microscopy revealed cells with condensed chromatin. The cytoplasm of these cells increased in electron density, the cisterns of rough endoplasmic reticulum were dilated and the cristae of mitochondria disappeared. These cells were comparable to the physiologically degenerative cells reported in other organs. These degenerative cells are joined to adjacent cells by desmosomes and are believed to be derived from the epithelial cells. Further, degenerative bodies, composed of amorphous structures and enveloped by a limiting membrane, were observed in the epithelial cells. They are presumed to be derived from the degenerated cells engulfed by neighboring cells. From this evidence, phagocytic ability is attributed to the epithelial cells of the tooth germ.

Animals↗
PubMed1977

Mitochondrial granule distribution in tooth germ cells.

Incisor and molar tooth germs of albino rats sacrificed at the eighteenth and twentieth days in utero and one to seven days after birth were studied with light and electron microscopy. Observations of the various stages of tooth development in molars established that intramitochondrial granules in odontoblasts were comparable to the intramitochondrial granules of other hard tissue cells. These electron-dense deposits appeared in mitochondria in an appreciable number only when odontoblasts become engaged in dentin mineralization. When dentin mineralization was advanced the odontoblast mitochondria appeared devoid of these deposits. Mesenchymal cells and preodontoblasts of the pulp were not involved in this activity.

Animals↗
PubMed1984

Autotransplantation and replantation of tooth germs in monkeys. Effect of damage to the dental follicle and position of transplant in the alveolus.

The effect of damage to the follicle and of superficial positioning of tooth germs after replantation or autotransplantation was studied in green Vervet monkeys. Radiographs were taken immediately after surgery and 3, 6 and 9 months postoperatively, whereafter the animals were sacrificed. The tissue blocks were sectioned in step serial sections along a frontal plane and histologic and radiographic evaluation was made. Tooth germs whose follicle was either damaged or removed showed no sign of eruption, while tooth germs which were replanted with an intact follicle were fully erupted within 3 months. Non-operated control teeth were fully erupted within 6 months. Histologic examination revealed that roots of replanted teeth were only slightly shorter than the non-operated control teeth. Transplantation of tooth germs in different positions in relation to the alveolar crest showed that teeth placed in their original position attained an average tooth length of 12.5 mm, whereas teeth placed in a semi-erupted position achieved an average length of only 9.7 mm. A control group of non-operated incisors demonstrated an average tooth length of 13.7 mm. This study indicates that damage to the follicle at the time of replantation of tooth germs is of major importance for tooth eruption and that placing tooth germs in a semi-erupted position adversely influences later root development.

Alveolar Process↗
PubMed1980

The effect of fluoride on ameloblasts of mouse molar tooth germs "in vitro".

Two day-old mouse first molar tooth germs were grown in medium containing sodium fluoride for up to 4 days. Dose levels ranged from 200 ppm fluoride down to 0.1 ppm. A dose of 100 ppm was lethal to tooth germs. Doses from 50 to 5 ppm resulted in consistent production of chains of large clear vacuoles in the supranuclear region of secretory ameloblasts together with large dark staining globules in the apical one third to one half of these cells. Enamel matrix secretion was arrested within this dose range as evidenced by the presence of only a very thin layer of enamel matrix in sections as compared with a thicker layer in similar areas of controls. Presecretory ameloblasts were not visibly affected by fluoride. Some variations in these effects were observed at 2 and 1 ppm fluoride. Tooth germs treated with 10 ppm for 48 hours, then grown in control medium for 4 days, underwent reversal of characteristic fluoride effects with resumption of enamel secretion. One ppm appeared to be the lower limit for production of these effects as observed with the light microscope. The cytology of fluorotic ameloblasts agreed with that of in vivo fluoride studies. Indications are that this in vitro system is promising for study of fluoride effects on developing teeth. Its advantages are discussed.

Ameloblasts↗
PubMed2002

The influence of parathyroid hormone-related protein (PTHrP) on tooth-germ development and osteoclastogenesis in alveolar bone of PTHrP-knock out and wild-type mice in vitro.

In a previous study, it was shown that tooth germs of neonatal homozygous parathyroid hormone-related protein (PTHrP)-knockout mice are penetrated or compressed by the surrounding alveolar bone, suggesting an important role for PTHrP in the formation and activation of osteoclasts around growing tooth germs. In order to elucidate the role of PTHrP during the development of the tooth germ and related structures, mandibular explants containing cap stage tooth germs of embryonic day 14, homozygous mice were here cultured with or without surrounding alveolar bone. There was no difference in the number of tartrate-resistant acid phosphatase-positive multinucleated osteoclastic cells around the first molars of homozygous and wild-type mice. After 10 days of culture, osteoclastic cells were rarely present in explants from homozygous mice and penetration of alveolar bone into the dental papilla was observed. The decline in osteoclast number was partly restored by the addition of PTHrP to the culture. Tooth germs of both wild-type and homozygous mice cultured without alveolar bone developed well, with no apparent structural abnormality; dentine formation was evident after 10 days. These data suggest that PTHrP is not required for the development of the tooth germ proper but is indispensable in promoting the osteoclast formation required to accommodate that development.

Acid Phosphatase↗
PubMed1991

[In vitro effect of chlorpromazine on the mineralization of tooth germ in mice--comparison with that of retinoic acid and HEBP].

Effects of chlorpromazine on the mineralization and alkaline phosphatase activity (ALP) in the tooth germ were examined and compared with those of retinoic acid and HEBP (1-hydroxyethylidene-1, 1-bisphosphonate). Mandibular first molars from 17-day-old mouse embryos were cultured with or without drugs. Calcium content and ALP in the tooth germ increased gradually from 0 to 7 days in culture, the increase of calcium being preceded by that of ALP. Retinoic acid suppressed increases of calcium and ALP in the tooth germ but not in the specimens precultured for 2 days, suggesting that retinoic acid inhibits the mineralization at an early developmental stage of the tooth. HEBP, a physiochemical inhibitor of mineralization, suppressed the increase of calcium, but significantly enhanced the increased of ALP in the tooth germ. Chlorpromazine, which has an antagonistic action towards calmodulin, also suppressed the increases of calcium and ALP in the tooth germ. Calmodulin antagonists W-7 and W-5 similarly suppressed the increases of calcium and ALP; W-5 had less effects on both calcium and ALP. These results indicate that calmodulin may be involved in the regulation of the mineralization in the tooth germ. These drugs are shown to possess different modes of inhibitory action on the mineralization.

Alkaline Phosphatase↗
PubMed1989

[In vitro cultivation of tooth germs (in mice)].

Mouse molars tooth buds on the bell stage were cultured, to investigate the best medium for their maintainance and their eventual clinical use. Tooth germs were cultivated during 3 to 8 days in three different medium: Eagle basal medium (liquid medium), agar-solidified medium and chick chorioallantoic membrane. The grafts were examined by light microscopy. Mesenchymal and atypical cells were counted in experimental and control groups. Our results showed that liquid medium was the best for 3-days buds cultures. Chorioallantoic membrane and agar-solidified medium showed better results for the maintainance of bud cultures for 8-day test. The objective of this study is to maintain in vitro tooth buds cultures for future transplants. This will also provide for the possibility of a more in-depth study of normal odontogenesis.

Animals↗
PubMed1989

Patterns of expression of intermediate filaments in ameloblastoma and human fetal tooth germ.

Monoclonal antibodies (Mab) were used to study the expression of cytokeratins and vimentin in various histological types of ameloblastoma and in human fetal tooth germ. The ameloblastoma and the tooth germ epithelia showed characteristics of both simple glandular and stratified squamous epithelial cells. Cytokeratin No. 18 was detected focally in most ameloblastomas studied but not in fetal odontogenic epithelia. Cytokeratins Nos. 8 and 19 were expressed in all epithelial elements of ameloblastomas and tooth germs. Only two tumors showed focally characteristics of keratinizing epithelia also seen in dental lamina but not in the enamel organ. All tumors except the granular cell ameloblastoma showed a variable coexpression of vimentin and cytokeratins in their neoplastic epithelia. A similar coexpression was detected in the stellate reticulum cells of the developing tooth. Ameloblastoma and human tooth germ epithelia share complex pattern of cytokeratin polypeptides together with coexpression of vimentin. The results strongly support the theory that ameloblastomas are of odontogenic origin and not direct derivatives of basal cells of oral epithelium or epidermis.

Adolescent↗
PubMed1987

The development of mandibular molar tooth germs isografted in the mouse spleen.

Tooth germs taken from 13-day-old embryonic mice and isografted in the spleen of adult mice were examined by light and electron microscopy. Well-organized tooth structures from the early cap stage to fully developed and mineralized mature teeth were obtained up to day 60 after transplantation. Germs on day 2 were similar to those prior to the onset of grafting but reached the late cap stage of development on day 4. On day 6, enamel and dentine formation were initiated and inner enamel epithelium and dental papilla cells were polarized. On days 10-15, enamel-matrix secretion was completed and almost all ameloblasts had become resorptive enamel epithelium. India ink injected from the recipient caudal vein accumulated to the capillaries within the pulp throughout the newly-formed vessels. On day 20, defined root formation had begun but occasionally irregular and cellular osteodentine was formed in root areas. On day 30, transplants were covered with reduced enamel epithelium and acellular cementum was formed at the root areas together with rudimentary periodontal ligament fibres. Cellular cementum became thicker up to day 40. There was little evidence of cellular infiltration from recipient tissue up to day 60. The spleen seems to be a suitable site for transplantation of tooth germs.

Animals↗
PubMed1985

Effects of maternal caffeine intake on the growth of rat tooth germs in protein-energy malnourished neonates.

Fourteen rat dams with 8 pups each were fed either a 6, 12 or 20 per cent protein diet upon birth. Another group of 12 dams with the same number of pups was pair-fed either a 6, 12 or 20 per cent protein diet supplemented with caffeine (2 mg/100 g body weight). At day 15, randomly-selected pups were injected with [14C]-proline to determine collagen synthesis of the incisor and molar tooth germs. Another group of pups was used to determine calcium content of these tooth germs. Body weight, incisor weight and total calcium contents of tooth germs of pups from dams fed with 6 per cent protein diet were greater in the caffeine-supplemented group, whereas in the 20 per cent protein diet with caffeine group, these parameters were lower. The molar weights of the 12 per cent protein diet with caffeine animals were greater than the 12 per cent group without caffeine. The total hydroxyproline content of the incisor tooth germs from animals in the 12 per cent protein diet with caffeine was greater than is the non-caffeine group. However, total hydroxyproline of the molar tooth germs in the 20 per cent protein groups with caffeine was less than in the non-caffeine group. The rate of collagen synthesis of the incisor and molar tooth germs showed no difference in the presence or absence of caffeine in the 6, 12 and 20 per cent protein groups. Incisor and molar tooth germs are thus affected differently by the interaction of protein and caffeine, possibly due to differences in the pattern of tooth development.

Animals↗
PubMed2004

[Establishment of a culture system of chick embryo for mouse tooth germ development].

OBJECTIVE: To establish a new culture system for mouse tooth germs in chick embryo. METHODS: The mandibular first molar germ fragments of 15 embryonic days' Kunming mouse embryo were implanted into the lateral mesenchyme of 4-5 days' chick embryo wing buds in ove. Eggs were reincubated and implanted tissues were examined by histochemistry. RESULTS: The cultured tooth germ development continued from cap stage to latest bell stage. The ameloblast and the odontoblast all differentiated maturely and secreted matrix. CONCLUSION: 4-5 days' wing buds chick embryo could serve as developing the mouse tooth germs and demonstrate well physiological process of differentiation and morphogenesis.

Ameloblasts↗
PubMed1996

Determination of enamel protein synthesized by recombined mouse molar tooth germs in organ culture.

Epithelial-mesenchymal interaction is a prerequisite for tooth morphogenesis. To study this interaction, inner enamel epithelium and dental papilla mesenchyme of molar tooth germs from a 16.5-day mouse embryo were dissociated enzymatically and cultured alone or after recombination. Characteristic matrix protein synthesized and secreted by recombined tooth germ was determined quantitatively by enzyme-linked immunosorbent assay. The protein was detected in the culture of recombined tooth germ but not of dissociated enamel epithelium alone. The amount of enamel protein increased until 8 days in culture. Morphological differentiation of the recombined epithelial rudiment into ameloblasts and enamel protein production were confirmed.

Ameloblasts↗
PubMed1989

[Production of monoclonal antibodies against tooth germs of hamster or rat by in vitro immunization].

The in vitro immunization procedure for the production of monoclonal antibodies has several advantages over the in vivo procedure; e.g. it requires a much smaller amount of immunogen and only a few days are required for immunization. However, no in vitro immunization procedure for the production of monoclonal antibodies against tooth germs has been reported. By means of in vitro immunization, we tried to produce monoclonal antibodies against the components of tooth germs which would be useful for immunohistochemical investigation of the development of tooth germs. Spleen cells of mice were immunized with a homogenate of molar tooth germs from hamsters or rats. The hybridomas produced were screened by immunohistochemical examination of paraffin sections of tooth germs. We obtained five monoclonal antibodies reacting with the dental tissues; ameloblasts, odontoblasts, enamel, dentin, predentin, and dental sac. One of them reacted specifically with only ameloblasts and one with ameloblasts and enamel, but the others are not specific for individual dental tissues. The results demonstrate that in vitro immunization is a satisfactory method for producing monoclonal antibodies useful for immunohistochemical investigation of tooth germs.

Animals↗