Ivan Míšek

Tooth-bone morphogenesis during postnatal stages of mouse first molar development.

The first mouse molar (M1) is the most common model for odontogenesis, with research particularly focused on prenatal development. However, the functional dentition forms postnatally, when the histogenesis and morphogenesis of the tooth is completed, the roots form and the tooth physically anchors into the jaw. In this work, M1 was studied from birth to eruption, assessing morphogenesis, proliferation and apoptosis, and correlating these with remodeling of the surrounding bony tissue. The M1 completed crown formation between postnatal (P) daysu20030–2, and the development of the tooth root was initiated at P4. From P2 until P12, cell proliferation in the dental epithelium reduced and shifted downward to the apical region of the forming root. In contrast, proliferation was maintained or increased in the mesenchymal cells of the dental follicle. At later stages, before tooth eruption (P20), cell proliferation suddenly ceased. This withdrawal from the cell cycle correlated with tooth mineralization and mesenchymal differentiation. Apoptosis was observed during all stages of M1 postnatal morphogenesis, playing a role in the removal of cells such as osteoblasts in the mandibular region and working together with osteoclasts to remodel the bone around the developing tooth. At more advanced developmental stages, apoptotic cells and bodies accumulated in the cell layers above the tooth cusps, in the path of eruption. Three‐dimensional reconstruction of the developing postnatal tooth and bone indicates that the alveolar crypts form by resorption underneath the primordia, whereas the ridges form by active bone growth between the teeth and roots to form a functional complex.

Morphogenesis and bone integration of the mouse mandibular third molar

The mouse third molar (M3) develops postnatally and is thus a unique model for studying the integration of a non-mineralized tooth with mineralized bone. This study assessed the morphogenesis of the mouse M3, related to the alveolar bone, comparing M3 development with that of the first molar (M1), the most common model in odontogenesis. The mandibular M3 was evaluated from initiation to eruption by morphology and by assessing patterns of proliferation, apoptosis, osteoclast distribution, and gene expression. Three-dimensional reconstruction and explant cultures were also used. Initiation of M3 occurred perinatally, as an extension of the second molar (M2) which grew into a region of soft mesenchymal tissue above the M2, still far away from the alveolar bone. The bone-free M3 bud gradually became encapsulated by bone at the cap stage at postnatal day 3. Osteoclasts were first visible at postnatal day 4 when the M3 came into close contact with the bone. The number of osteoclasts increased from postnatal day 8 to postnatal day 12 to form a space for the growing tooth. The M3 had erupted by postnatal day 26. The M3, although smaller than the M1, passed through the same developmental stages over a similar time span but showed differences in initiation and in the timing of bone encapsulation.

Three-dimensional reconstruction studies and morphometric analysis of rudimental tooth primordia in the upper incisor region of the sheep (Ovis aries, Ruminantia)

The functional dentition of the domestic sheep lacks all upper incisors and the upper canines. Nevertheless, occurrence of a dental lamina and rudimental tooth primordia had been described in the upper incisor region of the sheep. The aim of this study was to describe temporo-spatial pattern of origin and regression of these rudimental tooth primordia by light microscopy, computer-aided three-dimensional reconstruction and morphometry of the dental epithelium. Transient existence of a dental lamina in the upper incisor region of the sheep and three epithelial thickenings on its deep mesenchymal margin has been observed at day of ontogeny (DO) 48-53. They could not been identified as full-value tooth primordia, because they did not induce differentiation of tooth mesenchyme, but they could represent last remnants of functional upper incisors in early ancestors of ruminants. Additionally, a large rudimental upper canine primordium near the sutura maxilloincisiva occurred at DO43, reached early cap stage at DO52 and started to regress at DO53. Thus, our findings showed a discrepancy between the embryonic and adult dental pattern in the sheep. Similar molecular mechanisms as described for diastemal tooth rudiments in rodents could be involved during regression of rudimental tooth primordia in the upper incisor region of the sheep.

The Effects of Two Different Decalcification Procedures on Size and Structure of Embryonic Epithelial Tissue in Objects Prepared for Light Microscopy

Electrolytic decalcification is a very fast and effective method for removing calcium compounds from bones with minimum damage to tissues. Changes of dimension of tissues in histological sections prepared from specimens decalcified by immersion in a formic acid solution and sections prepared from specimens treated in an electrolytic decalcifier were studied. Heads of mouse foetuses were cut in half, decalcified by one of the above‐mentioned methods and embedded in histowax. Dimensional changes of skin, tongue and nasal epithelia in histological sections were evaluated by t‐test. Significant shrinking and other unwanted effects of decalcification, such as acidophilia of nuclei, were found in objects decalcified by both methods. No significant differences in the effects of the two methods on tissue dimensions were demonstrated. It is concluded that both decalcification methods are equivalent from the qualitative point of view.

A Striated Muscle on the Hard Palate of Rodents and Rabbits

A striated muscle of the hard palate has been previously described in some rodents and rabbits. It is not termed in the official veterinary anatomical nomenclature. The aim of this work was to verify the existence of this muscle. Heads of the golden hamster (Mesocricetus auratus), the guinea pig (Cavia aperea f. porcellus), the laboratory rat (Rattus norvegicus var. alba), the field vole (Microtus agrestis) and the domestic rabbit (Oryctolagus cuniculus f. domesticus) have been dissected. Moreover, histological sections have been prepared from heads of the field vole. In all species under study, we could detect a striated muscle of the hard palate composed of an anterior and a posterior muscle. The anterior muscle originated on the os incisivum and diverged in anterior, lateral and posterior directions. The posterior muscle originated on the processus palatinus maxillae and verged into the m. buccinator. Inter‐species differences could be detected in shape and position of the muscle. The palatal muscle was innervated by the ramus buccalis of the facial nerve. Whether this muscle should be classified as an individual facial muscle or as a part of the m. buccinator remains to be discussed.

The lateral enamel lamina--component of tooth primordia in selected mammalian species.

The lateral enamel lamina (LEL) is a part of the enamel organ, which is probably not involved in tooth formation. It represents, besides the stalk of the tooth primordium, a second interconnection between enamel organ and oral epithelium or vestibular lamina. We detected the LEL in the sheep ( Ovis aries ), the dolphin ( Stenella attenuata ), and the vole ( Microtus agrestis ) by light microscopy and computer-aided three-dimensional reconstruction. The LEL could be found in cap to bell stage tooth primordia, most clearly in slowly developing tooth germs. LEL-like structures have been furthermore described or depicted in tooth germs of the mouse, the elk ( Alces alces ), the dugong ( Dugong dugong ), the elephant ( Loxodonta africana ), and the human. Probably it is a part of all mammalian tooth primordia that undergoes regression during morphogenesis of the enamel organ. As a reducing structure, it should be considered in studies of tooth development.