Local Irradiation of Mouse Tooth Germ Gives Insight into the Direct Effects of Irradiation on Root Development.

To elucidate the mechanism underlying the failure of root formation after irradiation, we established a method of local irradiation of the molar tooth germ and demonstrated that radiation directly affected dental root development. In the current study, to locally irradiate the lower first molars of 5-day-old C57BL/6J mice, we used lead glass containing a hole as a collimator. We confirmed that our local irradiation method targeted only the tooth germ. The irradiated root was immature in terms of apical growth, and dentin formation was irregular along the outside of the root apices. Moreover, calcified tissue apically surrounded Hertwig's epithelial root sheath, which disappeared abnormally early. This method using a local irradiation experimental model will facilitate research into radiation-induced disorders of dental root formation.

Dose to the developing dentition during therapeutic irradiation: organ at risk determination and clinical implications.

PURPOSE: Irradiation of pediatric facial structures can cause severe impairment of permanent teeth later in life. We therefore focused on primary and permanent teeth as organs at risk, investigating the ability to identify individual teeth in children and infants and to correlate dose distributions with subsequent dental toxicity. METHODS AND MATERIALS: We retrospectively reviewed 14 pediatric patients who received a maximum dose >20 Gy(relative biological effectiveness, RBE) to 1 or more primary or permanent teeth between 2003 and 2009. The patients (aged 1-16 years) received spot-scanning proton therapy with 46 to 66 Gy(RBE) in 23 to 33 daily fractions for a variety of tumors, including rhabdomyosarcoma (n=10), sarcoma (n=2), teratoma (n=1), and carcinoma (n=1). Individual teeth were contoured on axial slices from planning computed tomography (CT) scans. Dose-volume histogram data were retrospectively obtained from total calculated delivered treatments. Dental follow-up information was obtained from external care providers. RESULTS: All primary teeth and permanent incisors, canines, premolars, and first and second molars were identifiable on CT scans in all patients as early as 1 year of age. Dose-volume histogram analysis showed wide dose variability, with a median 37 Gy(RBE) per tooth dose range across all individuals, and a median 50 Gy(RBE) intraindividual dose range across all teeth. Dental follow-up revealed absence of significant toxicity in 7 of 10 patients but severe localized toxicity in teeth receiving >20 Gy(RBE) among 3 patients who were all treated at <4 years of age. CONCLUSIONS: CT-based assessment of dose distribution to individual teeth is feasible, although delayed calcification may complicate tooth identification in the youngest patients. Patterns of dental dose exposure vary markedly within and among patients, corresponding to rapid dose falloff with protons. Severe localized dental toxicity was observed in a few patients receiving the largest doses of radiation at the youngest ages; however, multiple factors including concurrent chemotherapy confounded the dose-effect relationship. Further studies with larger cohorts and appropriate controls will be required.

Mandibular growth and tooth eruption after localized x-radiation.

The effect of localized x-radiation on the growth of mandibular bone and molar eruption was evaluated by morphometric methods. A dose of 20 Gy of x-radiation was given to the molar zone of growing rats. The animals were then killed in groups at 30 and 60 days postirradiation. Two groups of nonirradiated, age-matched rats were used as controls. Parameters related to molar eruption, mandibular length, and mandibular height were measured on lateral radiographs. The results obtained showed that the values of the biometric parameters were lower in experimental than in control animals. Odontoblastic atrophy, alveolodentary ankylosis, and meager or no root formation were the most conspicuous histologic findings. Osteodentin was found between canaliculary dentin and bone in cases of ankylosis. The morphometric data presented confirm the probability of alterations in mandibular growth and tooth eruption following x-radiation and suggest that this be considered in planning radiotherapy in children.

Effects of gamma radiation on incisor development of the prenatal albino mouse.

Fetuses of pregnant albino mouse exposed to 400 rad of gamma-irradiation, on the 12th gestational day, were compared with unirradiated fetuses to asses the radiation effect on developing incisors. Pregnant animals were sacrificed on day 18 post coitum, and their fetuses were decapitated. Heads were routinely prepared, frontally sectioned, and stained with hematoxylin and eosin. Histologic examination demonstrated that the development of the maxillary and mandibular incisors was retarded in all the experimental fetuses and were in early bell stage, whereas those of the control animals were elaborated their matrices. It was concluded that gamma-irradiation interferes with cytodifferentiation of the enamel organ and dental papilla and subsequently inhibits normal odontogenesis.

Ultrastructural study of osteodentin formation induced by irradiation.

A single dose of 16,000 rads was delivered to a 1 X 1.5 cm area of the cheek of 2 day old Wistar rats. The lower first molar region was removed 1, 3, 5 and 8 days postirradiation and processed for light and electron microscopy observation. The most distinct structural feature at optical level was osteodentin production in the dental papilla, which began at the level of the initiation of Hertwig's epithelial sheath and continued until the pulpal area was completely filled by osteodentin. At the ultrastructural level, ameloblasts exhibited large lipidic vacuoles and irregular intracytoplasmic secretory granules. Odontoblasts underwent multiple regressive alterations with vacuolization and numerous myelin figures. Pulpal cells exhibited a capacity for odontoblastic differentiation with formation of one or more odontoblastic processes and dentin matrix secretion. Thus, irradiation resulted in stimulation of a great potential for dentin production in these cells.

SDH activity in the developing incisors or irradiated mouse fetuses.

Pregnant CD1 Swiss albino mice were irradiated with 400 rads of whole body X-irradiation on the twelfth gestational day. The animals were then sacrificed beginning on day 14 through 20 of gestation by chloroform inhalation. The fetuses were extirpated via laparotomy and decapitated. The severed heads were rapidly frozen and sectioned in a cryostat. The sections were affixed to glass slides and incubated for succinic dehydrogenase activity according to the method of Nachlas et al. (1957) and counterstained in Safranin 0, routinely dehydrated and mounted. Data from observations indicated that siccinic dehydrogenase activity appeared normal in the tissue layers of the developing tooth germ when compared to control animals. When the experimental procedure had invoked damage to the developing tooth, succinic dehydrogenase activity was lessened relative to the degree of damage. Presumably the X-irradiation had affected the cellular maturation process thereby reducing the functional competency of the cells as illustrated by the reduced enzyme activity.

Sensitivy of mouse molar tooth germs to x-ray irradiation in vitro.

Molar tooth germs, extirpated from 18-day mouse fetuses were cultured on Millipore filter strips in Falcon organ culture dishes. The tooth germs were exposed to 250 kVcp X-rays at 106 R/min. for a total exposure of 1 600 R. Tissues were harvested on a daily basis for a total period of 12 days and were examined microscopically, utilizing H and E stain. Severe disorganization of the tooth germs was evident within 24 hours of irradiation. The basement membrane became hyalinized; pyknotic nuclei and lysed cells were observed throughout the dental papilla, but mostly in the regions of the presumptive cusps. Although a thin layer of predentin was elaborated by the odontoblasts, the matrix failed to calcify and enamel matrix was not produced. Cultures older than 10 days demonstrated extensive cell death. The entire pulp was reduced to a mass of necrotic cells and the ameloblastic layer consisted of an epithelial remnant covering the cuspal tips.

The effect of irradiation on developing teeth.

Although the consequences of ionizing irradiation on adult teeth have been adequately described in the literature, there have been few reports concerning the adverse effects on the developing dentition. A case is presented in which the jaws were irradiated at an early age and it was possible to examine some of the abnormal teeth in detail by using clinical, radiographic, and histologic techniques.

[The problem of damage to the tooth germ by ionizing radiation]. / Zur Frage der Zahnkeimschädigung durch ionisierende Strahlen.

Experiments on animals (four-week-old dogs) were conducted in an investigation made to study the possibility of dose-dependent tooth-germ damage produced by ionizing radiation. The individual doses were 50 r and 200 r, respectively, and they were administered once to three times at weekly intervals. Hyperemia and edemata could be observed on tooth-germ pulps from 150 r onward. Both of these conditions became more acute as the radiation dose increased (from 150 r to 600 r). Possible damage to both the dentin and enamel is pointed out.