Effect of hematoma on early degradation behavior of magnesium after implantation.

The corrosion of magnesium (Mg)-based bioabsorbable implanting devices is influenced by implantation environment which dynamically changes by biological response including wound healing. Understanding the corrosion mechanisms along the healing process is essential for the development of Mg-based devices. In this study, a hematoma model was created in a rat femur to analyze Mg corrosion with hematoma in the early stage of implantation. Pure Mg specimen (99.9%,ϕ1.2 × 6 mm) was implanted in rat femur under either hematoma or non-hematoma conditions. After a designated period of implantation, the specimens were collected and weighed. The insoluble salts formed on the specimen surfaces were analyzed using scanning electron microscopy, energy-dispersive x-ray spectroscopy, and Raman spectroscopy on days 1, 3, and 7. The results indicate that hematomas promote Mg corrosion and change the insoluble salt precipitation. The weight loss of the hematoma group (27.31 ± 5.91 µg mm-2) was significantly larger than that of the non-hematoma group (14.77 ± 3.28 µg mm-2) on day 7. In the non-hematoma group, carbonate and phosphate were detected even on day 1, but the only latter was detected on day 7. In the hematoma group, hydroxide was detected on day 1, followed by the formation of carbonate and phosphate on days 3 and 7. The obtained results suggest the hypoxic and acidic microenvironment in hematomas accelerates the Mg corrosion immediately after implantation, and the subsequent hematoma resorption process leads to the formation of phosphate and carbonate with organic molecules. This study revealed the risk of hematomas as an acceleration factor of the corrosion of Mg-based devices leading to the early implant failure. It is important to consider this risk in the design of Mg-based devices and to optimize surgical procedures controlling hemorrhage at implantation and reducing unexpected bleeding after surgery.

In vitroanalysis of insoluble salt formation mechanism associated with Mg corrosion-variations depending on the diffusion environment in model tissue.

Magnesium (Mg) alloys have attracted attention as biodegradable metals, but the details of their corrosion behavior under biological environment have not been elucidated. Previous studies have suggested that diffusion through blood flow may influence Mg corrosion. Therefore, to understand the degradation behaviors of Mg, we analyzed insoluble salt precipitation associated with Mg corrosion in model tissue with different diffusion rates. A pure Mg specimen was immersed into a model tissue prepared with cell culture medium supplemented by a thickener at a different concentration (0.2%-0.5%) to form the gel. Micro-focus x-ray computed tomography of the gel was performed to observe gas cavity formation around the specimen. The insoluble salt layer formed on the specimen surface were analyzed by scanning electron microscopy with energy-dispersive x-ray spectroscopy, and Raman spectroscopy. As results, gas cavity formation was observed for all specimens. At day 7, the gas cavity volume was the highest at 0.5% thickener gel followed by 0.3% thickener gel. The insoluble salts were classified into three types based on their morphology; plate-like, granular-like, and crater-like salts. The crater-like salts were observed to cover 16.8 ± 3.9% of the specimen surface immersed in the 0.5% thickener gel, at the specimen area contacted to the gas cavity. The crater-like salts were composed by Mg hydroxide and carbonate from the deepest to the top layer. In plate-like or granular-like salts, Mg carbonate was formed in the deepest layer, but phosphates and carbonates, mainly containing calcium not Mg, were formed on the surface layer. In conclusion, the increase in the thickener concentration increased the gas cavity volume contacting to the specimen surface, resulting in the increase in precipitation of Mg hydroxide and carbonate, composing crater-like salts. Mg hydroxide and carbonate precipitation suggests the local increase in OH-concentration, which may be attributed to the decrease in diffusion rate.

Estimation of external dose for wild Japanese macaques captured in Fukushima prefecture: decomposition of electron spin resonance spectrum.

The signal intensities of CO2- radicals in teeth can be utilised as an individual indicator of the cumulative external dose for animals. To accurately determine the external dose, it is desirable to analyse the CO2- radical intensity and improve its detection limit. We recently reported a dose-response in the range of 0-200 mGy and estimated the absorbed dose for seven wild Japanese macaques captured in/around the related areas to the Fukushima Daiichi Nuclear Power Plant accident. Herein, for further improvement of this method, we examined the electron spin resonance spectra of the teeth of these seven and an additional four macaques captured in Fukushima by applying two spectrum-decomposition algorithms.

Detection limit of electron spin resonance for Japanese deciduous tooth enamel and density separation method for enamel-dentine separation.

Electron spin resonance (ESR) dosimetry is one of the most powerful tools for radiation dose reconstruction. The detection limit of this technique using human teeth is reported to be 56 mGy or 67 mGy; however, the absorbed dose of Fukushima residents after the Fukushima Daiichi Nuclear Power Plant (FNPP) accident was estimated to be lower than this detection limit. Our aim is to assess the absorbed radiation dose of children in Fukushima Prefecture after the accident; therefore, it is important to estimate the detection limit for their teeth. The detection limit for enamel of deciduous teeth of Japanese children separated by the mechanical method is estimated to be 115.0 mGy. The density separation method can effectively separate enamel from third molars of Japanese people. As we have collected thousands of teeth from children in Fukushima, the present technique may be useful to examine their external absorbed dose after the FNPP accident.

Biodegradation behaviors of magnesium(Mg)-based alloy nails in autologous bone grafts: In vivo study in rabbit skulls.

OBJECTIVE: In this study, autologous bone grafts using bone-fixing nails made of magnesium-zinc-calcium ternary alloys were performed using rabbit skulls. MATERIAL AND METHODS: Two types of nails for bone fixation were prepared: 2.5 mm width, 3 mm length and 2.5 mm width, 2 mm length. A disk-shaped bone with a diameter of 5 mm was resected from the parietal bone and fixed with a 3 mm long nail. As a control group, a 2 mm long nail was driven into the existing bone. The rabbits were sacrificed at 1, 4, 12, and 24 weeks after surgery. The resected samples were observed with micro X-ray CT, and embedded in methyl methacrylate to prepare non-decalcified specimens. The in vivo localization of elements was examined using energy-dispersive X-ray spectroscopy (EDS). RESULTS: Micro X-ray CT images of samples showed volume reduction due to degradation in both the bone graft and control groups. No significant difference in the amount of degradation between the two groups was observed, however characteristic degradation processes were observed in each group. The samples stained with alizarin red S showed amorphous areas around the nails, which were considered as corrosion products and contacted directly with the newly formed bones. EDS analysis showed that corrosion products were mainly composed of magnesium and oxygen at an early stage, while calcium and phosphorus were detected on the surface layer during the long-term observation. CONCLUSIONS: The degradation speed of the magnesium alloy nails varied depending on the shapes of the nails and surrounding tissue conditions. A calcium phosphate layer was formed on the surface of magnesium alloy nails, suggesting that the degradation rate of the nail was slow.

A collagen membrane for periosteal expansion osteogenesis using a timed-release system in rabbit calvaria.

BACKGROUND: The purpose of this study was to evaluate the effects of resorbable membranes, combined with a shape memory alloy (SMA) mesh device, on bone formation using a timed-release system for periosteal expansion osteogenesis (TIME-PEO). MATERIALS AND METHODS: Twelve Japanese white rabbits were used in this study. An SMA device was inserted under the forehead periosteum, pushed and bent for attachment to the bone surface, and then fixed using resorbable thread. The rabbits were divided into four groups: C1 (5 weeks postoperatively without membrane), C2 (8 weeks postoperatively without membrane), E1 (5 weeks postoperatively with membrane), and E2 (8 weeks postoperatively with membrane). The rabbits were killed 5 or 8 weeks after the operation and the newly formed bone was assessed histologically and radiographically. RESULTS: SMA devices, concealed under soft tissue until the time of euthanasia, did not cause active inflammation. The mean activation height, from the original bone surface to the midpoint of the mesh, was 3.1 ± 0.6 mm. Newly formed bone was observed, and most of the subperiosteal space underneath the device was occupied by fibrous tissue. Immature bone was present at the outer surface of the original skull bone in all groups. On histomorphometric analysis, there was no significant difference in the volume of the new bone between C1 and E1 (p = 0.885), and C2 and E2 (p = 0.545). CONCLUSIONS: PEO using an SMA mesh device, which is based on guided bone regeneration (in atrophic alveolar bone), shows promise as an alternative for bone augmentation, irrespective of whether a resorbable membrane is used.

Environmental radiation on large Japanese field mice in Fukushima reduced colony forming potential in hematopoietic progenitor cells without inducing genomic instability.

PURPOSE: To study the environmental radiation effects of wild animals after the Fukushima Dai-ichi nuclear power plant accident, we assessed effects on hematopoietic progenitor cells (HPCs) in large Japanese field mice (Apodemus speciosus). MATERIALS AND METHODS: A. speciosus were collected from three contaminated sites and control area. The air dose-rates at the control and contaminated areas were 0.96 ± 0.05 µGy/d (Hirosaki), 14.4 ± 2.4 µGy/d (Tanashio), 208.8 ± 31.2 µGy/d (Ide), 470.4 ± 93.6 µGy/d (Omaru), respectively. We investigated possible DNA damage and pro-inflammatory markers in the bone marrow (BM) cells. The colony-forming potential of BM cells was estimated by the number of HPC colony-forming cells. Radiation-induced genomic instability (RIGI) in HPCs was also analyzed by quantifying delayed DNA damage in CFU-GM clones. RESULTS: Although no significant differences in DNA damage and inflammation markers in BM cells from control and contaminated areas, the number of HPC colonies exhibited an inverse correlation with air dose-rate. With regard to RIGI, no significant differences in DNA damage of CFU-GM clones between the mice from the control and the three contaminated areas. CONCLUSIONS: Our study suggests that low dose-rate radiation of more than 200 Gy/d reduced HPCs, possibly eliminating genomically unstable HPCs.

Radioactivity and radionuclides in deciduous teeth formed before the Fukushima-Daiichi Nuclear Power Plant accident.

The Fukushima-Daiichi Nuclear Power Plant (FNPP) accident in March of 2011 released substantial amounts of radionuclides into the environment. We collected 4,957 deciduous teeth formed in children before the Fukushima accident to obtain precise control data for teeth formed after the accident. Radioactivity was measured using imaging plates (IP) and epidemiologically assessed using multivariate regression analysis. Additionally, we measured 90Sr, 137Cs, and natural radionuclides which might be present in teeth. Epidemiological studies of IP showed that the amount of radioactivity in teeth from Fukushima prefecture was similar to that from reference prefectures. We found that artificial radionuclides of 90Sr and 137Cs, which were believed to have originated from past nuclear disasters, and natural radionuclides including 40 K and daughter nuclides in the 238U and 232Th series contributed to the generation of radioactivity in teeth. We also found no evidence to suggest that radionuclides originating from the FNPP accident significantly contaminated pre-existing teeth. This is the first large-scale investigation of radioactivity and radionuclides in teeth. The present findings will be indispensable for future studies of teeth formed after the FNPP accident, which will fall out over the next several years and might be more contaminated with radionuclides.

Wound healing effect of autologous fibrin glue and polyglycolic acid sheets in a rat back skin defect model.

Fibrin glue from autologous plasma may prevent viral infection and allergic reaction. Moreover, this biomaterial contains growth factors such as TGF-ß and VEGF that promote reconstruction of the mucous membrane by stimulating fibroblast proliferation and angiogenesis. Thus, autologous fibrin glue is predicted to improve healing better than commercial fibrin glue. Here, we evaluated the effects of autologous fibrin glue on the crucial early phase of wound healing. Epithelial defects were introduced in rats and covered with polyglycolic acid (PGA) sheets with or without commercial or autologous fibrin glue. Wound healing was assessed for six weeks by histology and immunohistochemistry. Our results demonstrate that wounds covered with PGA sheets and autologous fibrin glue achieved efficient wound healing without complications such as local infection or incomplete healing. The rate of recovery of the regenerating epithelium in this group was superior to that in wounds covered with PGA sheets and commercial fibrin glue. Immunohistochemistry of laminin, cytokeratin, and VEGF confirmed fine and rapid epithelial neogenesis. Collectively, our results indicate that covering surgical wounds with autologous fibrin glue promotes wound healing and epithelialization, improves safety, and reduces the risks of viral infection and allergic reaction associated with conventional techniques.

In vivo behaviors of highly flexible paper consisting of ultralong hydroxyapatite nanowires.

This study was conducted in order to investigate biological compatibility of a thin and flexible hydroxyapatite (HAP) paper which consists of ultralong hydroxyapatite nanowires. Circular-shaped cranial bone defects with a diameter of 8.8 mm were prepared to expose the dura maters in Wistar rats. The similar-sized, circular-shaped HAP paper was placed at the bottom of the bone defects. After 2, 4, and 8 weeks, the rats were sacrificed, and the experimental sections were examined by micro-CT scanning and histological observation. The HAP paper covered with fibrous tissues showed no inflammatory cell infiltration, and their thicknesses decreased over time. Tartrate-resistant acid phosphatase-positive osteoclast-like cells were induced around the edges of the HAP paper along with the exfoliation of the HAP paper. The newly-formed bones were observed in the bone-defected areas, either with a direct contact with the HAP paper or through thin fibrous tissues. The HAP paper-induced osteoblast differentiation was confirmed since the alkaline phosphatase activities were detected on the surfaces of the HAP paper. These results indicated that the HAP paper may induce osteogenesis without causing any harmful effects. The highly flexible HAP paper can contribute to further development of bone regenerative therapy.

Osteogenic response under the periosteum by magnesium implantation in rat tibia.

This study was designed to examine osteoconductive effects of Mg in rats tibia. The animals were sacrificed after 1, 2, and 8 weeks. The elemental analysis was performed using SEM/EDX at week 1. Following X-ray micrography at weeks 2 and 8, samples were embedded in paraffin. The expression of osteocalcin was observed by immunohistochemical staining. The element concentrations of fibrous capsules around the specimens were also measured by ICP-MS. The concentrations of Ca and P on the surface of the Mg specimen increased in SEM/EDX. The tissue specimen showed new bone formation on the bone surface near the implanted area. The concentrations of Mg, Ca, and P were high in the fibrous capsules surrounding Mg. Implantation induced differentiation of osteoblasts, and this process was considered to be associated with new bone formation. Induction of cell differentiation may be influenced by corrosion products in addition to corroding magnesium.

Periosteal expansion osteogenesis using an innovative, shape-memory polyethylene terephthalate membrane: An experimental study in rabbits.

Periosteal expansion osteogenesis (PEO) results in the formation of new bone in the gap between periosteum and original bone. The purpose of this study is to evaluate the use of a polyethylene terephthalate (PET) membrane as an activation device. A dome-shaped PET membrane coated with hydroxyapatite/gelatin on the inner side was inserted between the elevated periosteum and bone at the rabbit calvaria. In the experimental group, the membrane was pushed, bent, and attached to the bone surface and fixed with a titanium screw. In control group, the membrane was only inserted and fixed with titanium screw at original shape under the periosteum. After 7 days, the screw was removed and the mesh was activated in the experimental group. Three animals per group with or without setting a latency period for activation were sacrificed at 3 and 5 weeks after surgery. Bone formation was evaluated via micro-computed tomography and determined by histomorphometric methods and histological evaluation. No PET membrane-associated complications were observed during this study. The quantitative data by the area and the occupation of newly formed bone indicated that the experimental group had a higher volume of new bone than the control group at 3 weeks after surgery. Histologically, bone formation progressed to areas adjacent to the cortical perforations; many sinusoidal vessels ran from the perforations to overlying fibrous tissue via the new bone. No bone or obvious inflammatory cells were observed over the membrane. The PET membrane has biocompatible device for PEO that induces a natural osteogenic response at the gap between the original bone and periosteum.

In vivo behavior of untreated and compressed concentrated growth factors as biomaterials in rabbits.

To characterize concentrated growth factors (CGFs) in vivo, we examined the degradation of implanted CGF in rabbits. Untreated CGF (U-CGF) and compressed CGF (C-CGF) were subcutaneously implanted into the dorsum. Histological analyses showed that the U-CGF and C-CGF induced very few inflammatory cells and that the U-CGF and C-CGF were subsequently degraded with dendritic invasion of granulation tissue. The C-CGF histopathologically remained for longer term than the U-CGF. Aggregated CD31+ and RAM11+ cells appeared in and around the implanted CGF. The number of macrophages and blood vessels in the CGF-implanted groups was greater than that in the sham group. There were more blood vessels in the U-CGF group than that in the C-CGF and sham group. We showed that CGF was degraded by macrophages in 4 weeks and enhanced angiogenesis with dendritically branching new capillaries. Therefore, the U-CGF and C-CGF can be clinically applied as a biomaterial inducing angiogenesis.

The process of magnesium ion modification of titanium surface and the sustained-release of magnesium ions from its surface.

This study explored modification of an alkaline heat treated titanium surface, using magnesium ions, to improve bone compatibility through the sustained release of magnesium ions. Pure titanium surface was first subjected to alkaline treatment using 5 M NaOH then modified with magnesium through immersion in magnesium chloride solution before heating in a furnace at 600°C for 1 h. Use of at least 0.01 M magnesium chloride solution for at least 0.5 min, leads to introduction of 1.7 to 2.3 at% magnesium at a distribution close to saturation on the titanium surface. The modified titanium surface sustained long term release of magnesium ions in acidic solution for more than 168 h. It was further demonstrated that the process of sustained release of magnesium ions is influenced by pH and can be triggered by lowering it from neutral to 3.

Expression of immunoregulatory molecules PD-L1 and PD-1 in oral cancer and precancerous lesions: A cohort study of Japanese patients.

OBJECTIVE: An association of the programmed cell death-1 (PD-1) and its ligand PD-L1 with various types of malignant tumors has been established. This study aimed to investigate the role of the PD-L1/PD-1 pathway in oral squamous cell carcinoma (OSCC) and oral epithelial precursor lesions (OEPL). MATERIALS AND METHODS: We examined 106 OSCC and 79 OEPL specimens for PD-L1 and PD-1 expression by immunohistochemistry. The results were compared with clinicopathological features of OSCC patients. RESULTS: In OSCC and OEPL specimens, PD-L1 expression was detected predominantly in epithelial or carcinoma cells, whereas PD-1 expression was found mainly in infiltrating or stromal lymphocytes. Seventy-two OSCC (67.9%) and 21 OEPL (26.6%) specimens were positive for PD-L1, and 73 OSCC (68.9%) and 23 OEPL (29.2%) specimens were positive for PD-1. PD-L1 and PD-1 expression levels were significantly different between OEPL and OSCC specimens (P < 0.001). There were significant positive correlations between PD-L1 and PD-1 expression in OEPL and OSCC specimens (P < 0.001). PD-L1 and PD-1 immunoreactivity was significantly associated with tumor size (P < 0.05). PD-L1 and PD-1 immunoreactivity in cases with advanced TNM staging was significantly higher than that in low staging cases (P < 0.01). There were significant correlations between PD-L1 and PD-1 expression in OSCC specimens and pathological variables such as stromal lymphocytic reaction (P < 0.05) and invasion depth (P < 0.01). CONCLUSION: PD-L1 and PD-1 immunohistochemical status may be related to carcinogenesis, tumor progression, and prognosis in oral epithelial lesions. Agents targeting PD-1 and PD-L1 might be useful for OSCC treatment.

In vitro and in vivo analysis of the biodegradable behavior of a magnesium alloy for biomedical applications.

The present study was designed to investigate the biodegradation behavior of Mg alloy plates in the maxillofacial region. For in vitro analysis, the plates were immersed in saline solution and simulated body fluid. For in vivo, the plates were implanted into the tibia, head, back, abdominal cavity, and femur and assessed at 1, 2, and 4 weeks after implantation. After implantation, the plate volumes and the formed insoluble salt were measured via micro-computed tomography. SEM/EDX analysis of the insoluble salt and histological analysis of the surrounding tissues were performed. The volume loss of plates in the in vitro groups was higher than that in the in vivo groups. The volume loss was fastest in the abdomen, followed by the head, back, tibia, and femur. There were no statistically significant differences in the insoluble salt volume of the all implanted sites. The corrosion of the Mg alloy will be affected to the surrounding tissue responses. The material for the plate should be selected based on the characteristic that Mg alloys are decomposed relatively easily in the maxillofacial region.

Haematological analysis of Japanese macaques (Macaca fuscata) in the area affected by the Fukushima Daiichi Nuclear Power Plant accident.

Several populations of wild Japanese macaques (Macaca fuscata) inhabit the area around Fukushima Daiichi Nuclear Power Plant (FNPP). To measure and control the size of these populations, macaques are captured annually. Between May 2013 and December 2014, we performed a haematological analysis of Japanese macaques captured within a 40-km radius of FNPP, the location of a nuclear disaster two years post-accident. The dose-rate of radiocaesium was estimated using the ERICA Tool. The median internal dose-rate was 7.6 µGy/day (ranging from 1.8 to 219 µGy/day) and the external dose-rate was 13.9 µGy/day (ranging from 6.7 to 35.1 µGy/day). We performed multiple regression analyses to estimate the dose-rate effects on haematological values in peripheral blood and bone marrow. The white blood cell and platelet counts showed an inverse correlation with the internal dose-rate in mature macaques. Furthermore, the myeloid cell, megakaryocyte, and haematopoietic cell counts were inversely correlated and the occupancy of adipose tissue was positively correlated with internal dose-rate in femoral bone marrow of mature macaques. These relationships suggest that persistent whole body exposure to low-dose-rate radiation affects haematopoiesis in Japanese macaques.

Prolonged release of bone morphogenetic protein-2 in vivo by gene transfection with DNA-functionalized calcium phosphate nanoparticle-loaded collagen scaffolds.

In the combination of scaffolds immersed in growth factor solutions, the release of growth factors mainly depends on scaffold degradation. However, the release of bone morphogenetic protein (BMP)-2 at an appropriate concentration during the stage of tissue regeneration would enhance bone regeneration. To achieve this condition, the present study was performed to investigate the effects of scaffolds combined with gene transfection using non-viral vectors. Nanohydroxyapatite-collagen (nHAC) scaffolds cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) or ascorbic acid/copper chloride, and a collagen scaffold (Terdermis®) were prepared, loaded with BMP-2-encoding plasmid DNA-functionalized calcium phosphate nanoparticles (CaP), naked plasmid DNA, or BMP-2 solution, and implanted in rats. The yield of released BMP-2 and its releasing period, respectively, were larger and longer from the scaffolds loaded with CaP than from those incubated with BMP-2 solution. In addition, the alkaline phosphatase activity induced by the CaP-loaded scaffolds was higher. Histological analysis showed that released BMP-2 could be observed on the macrophages or multinuclear giant cells surrounding the nHAC fragments or collagen fibres. TRAP-positive or OCN-positive sites were observed in all groups and a mineralization area was observed in the Terdermis®/CaP sample. The present study demonstrates that gene transfection by scaffold loaded with CaP gene transfer vectors induces a larger yield of BMP-2 for a longer period than by scaffolds loaded with BMP-2 solution or naked plasmid.

Initial organ distribution and biological safety of Mg2+ released from a Mg alloy implant.

Magnesium (Mg) alloys are considered promising materials for biodegradable medical devices; however, the initial effects and distribution of released Mg2+ ions following implantation are unclear. This is addressed in the present study, using two types of Mg alloys implanted into rats. An in vitro immersion test was first carried out to quantify Mg2+ ions released from the alloys at early stages. Based on these data, we performed an in vivo experiment in which large amounts of alloys were subcutaneously implanted into the backs of rats for 1, 5, 10, and 25 h. Mg2+ accumulation in organs was measured by inductively coupled plasma mass spectrometry. In vivo, blood and urine Mg2+ concentrations were higher in rats receiving the implants than in controls after 1 h; however, the levels were within clinically accepted guidelines. The Mg2+ concentration in bone was significantly higher in the 25 h implanted group than in the other groups. Our results suggest that homeostasis is maintained by urinary excretion and bone accumulation of released Mg2+ ions in response to sudden changes in Mg2+ ion concentration in the body fluid in a large number of Mg alloy implants at the early stages.

90Sr specific activity of teeth of abandoned cattle after the Fukushima accident - teeth as an indicator of environmental pollution.

90Sr specific activity in the teeth of young cattle that were abandoned in Kawauchi village and Okuma town located in the former evacuation areas of the Fukushima-Daiichi Nuclear Power Plant (FNPP) accident were measured. Additionally, specific activity in contaminated surface soils sampled from the same area was measured. (1) All cattle teeth examined were contaminated with 90Sr. The specific activity, however, varied depending on the developmental stage of the teeth during the FNPP accident; teeth that had started development before the accident exhibited comparatively lower values, while teeth developed mainly after the accident showed higher values. (2) Values of 90Sr-specific activity in teeth formed after the FNPP accident were higher than those of the bulk soil but similar to those in the exchangeable fraction (water and CH3COONH4 soluble fractions) of the soil. The findings suggest that 90Sr was incorporated into the teeth during the process of development, and that 90Sr in the soluble and/or leachable fractions of the soil might migrate into teeth and contribute to the amount of 90Sr in the teeth. Thus, the concentration of 90Sr in teeth formed after the FNPP accident might reflect the extent of 90Sr pollution in the environment.