Fructooligosaccharide consumption improves the decreased dentin formation and mandibular defects following gastrectomy in rats.

OBJECTIVE: We examined the effects of fructooligosaccharides (FOS) consumption on gastrectomy-evoked osteopenia and disorders of dentin formation in rats. MATERIALS AND METHODS: Male Sprague-Dawley rats (n = 28, 35-day old) were equally divided into two groups; sham-operated and gastrectomized, and sham-operation or total gastrectomy was performed. Four weeks after each surgery, the rats were divided into two sub-groups (n = 7 each); with or without 7.5% FOS-feeding for 6 weeks. Backscattered electron images of the mandibular sections were taken to calculate trabecular bone area, cortical bone area and total scan area. Thereafter, the dentin formation rate in maxilla were calculated using a fluorescent microscope. RESULTS: Trabecular bone area and cortical bone area in GX rats were markedly decreased. FOS-feeding significantly counteracted this reduction, but not to the level seen in sham-operated rats. Total scan area in gastrectomized groups was significantly decreased. The dentin formation rate was not statistically different among the groups, except the gastrectomized group. CONCLUSION: These results suggest that FOS consumption partially restored osteopenia and almost completely restored the reduction in dentin formation following gastrectomy in rats.

Amelioration of 2,4,6-trinitrobenzene sulphonic acid induced colitis in angiotensinogen gene knockout mice.

BACKGROUND: A number of recent studies have demonstrated a protective effect of renin-angiotensin system (RAS) antagonism against immune mediated diseases such as myocarditis, chronic allograft rejection, and antiglomerular basement membrane nephritis. To our knowledge, there has been no report on the immunological contribution of the RAS in colonic tissue. AIMS: We evaluated the direct effect of angiotensin II (AII) on the pathogenesis of immune mediated colitis using angiotensinogen deficient homozygous (Atg-/-) mice. SUBJECTS: 2,4,6-Trinitrobenzene sulphonic acid (TNBS) colitis was induced in Atg-/- and wild-type (Atg+/+) mice. METHODS: Levels of proinflammatory cytokines in the colon were determined by enzyme linked immunosorbent assay. Histological analysis was performed simultaneously. RESULTS: Although Atg-/- mice developed colitis, the degree was much milder than that in Atg+/+ mice (p<0.05). Colonic cytokine analysis showed that the production of proinflammatory cytokines (interleukin (IL)-1beta, interferon gamma (IFN-gamma)) was impaired in Atg-/- mice. Furthermore, expression of cytokines such as IL-4 and IL-10 in the colon was predominant in Atg-/- compared with Atg+/+ mice after TNBS instillation (p<0.005, p<0.01, respectively). Similarly, subcutaneous infusion of losartan suppressed colitis (p<0.05) and the production of proinflammatory cytokines (IL-1beta, IFN-gamma). These results indicate that the RAS is directly involved in the pathogenesis of TNBS colitis through regulation of proinflammatory and anti-inflammatory cytokines in the colon. CONCLUSIONS: This study revealed that the RAS is involved in the immune system in the colon. Antagonism of the RAS is a potential prophylactic strategy for the treatment of human inflammatory bowel disease.

Developmental and functional analyses of CD8(+) NK1.1(+) T cells in class-I-restricted TCR transgenic mice.

Using a class-I-restricted T cell receptor (TCR) transgenic mice (Tgm), 2C (Valpha3.1/Vbeta 8.2, specific for L(d) + LSPFPFDL), the development and cytokine production of tg-TCR(+) NKT cells were analyzed. We found that CD8(+) or double negative (DN) NKT cells constituted a major population of NKT cells in the H-2(b/b) 2C Tgm (positive selecting background) or the H-2(b/d) 2C Tgm (negative selecting background), respectively. Virtually no NKT cells were generated in the H-2(k/k) 2C Tgm (neutral selecting background). CD8(+) NKT cells in the H-2(b/b) 2C Tgm expressed CD8alphabeta heterodimers, whereas those in the H-2(b/d) 2C Tgm expressed CD8alphaalpha homodimers. These findings suggest that development of a subpopulation of NKT cells is influenced by the H-2 molecules. Upon stimulation with anti-CD3 mAb, tg-TCR(+) NKT cells generated in the H-2(b/b) and H-2(b/d) backgrounds produced IFN-gamma, but not IL-4.

Immunohistochemical characterization of hepatoblastomas in B6C3F1 mice treated with diethylnitrosamine and sodium phenobarbital.

Hepatoblastomas (HBs) were induced in B6C3F1 male mice by diethylnitrosamine (DEN) and sodium phenobarbital (PB). Six-week-old mice received a single intraperitoneal dose of DEN followed by a continuous treatment with PB in diet at a concentration of 0 (group 1) or 500 (group 2) ppm for 50 weeks. HBs were observed in 13 of 21 (62%) group 2 mice, with typical histologic features as reported previously, while no such tumors were observed in group 1. Seven of 13 (54%) HBs were found in and/or adjacent to hepatocellular adenomas (HCAs) or hepatocellular carcinomas (HCCs). Immunohistochemically, all HBs were positive for S-100 protein but negative for keratin, alpha-fetoprotein (AFP), albumin (ALB) and vimentin, while HCC cells occasionally reacted positively for AFP with a mosaic pattern. HCC and HCA cells were occasionally positive for ALB. Non-neoplastic hepatocytes and normal bile ducts were positively stained for ALB and keratin/S-100 protein, respectively. S-100 protein is known to be expressed in many mesenchymal tissues and neoplasms including neuroectodermal elements but negative in cells of the hepatic lineage. Thus, the present immunohistochemical results suggested that mesenchymal differentiation occurs in mouse HB cells as observed in human HBs, one of the most frequent infant liver tumors in humans. Although the susceptibility of mouse HBs to PB-promotion suggests a hepatocytic histogenesis, the present immunohistochemical results support the hypothesis that the mouse HB is derived from pluripotent endodermal stem-like cells.

Details of true calcium absorption and calcium excretion into feces in female rats during growth.

The details of calcium flux during growth were examined in female rats. Twenty-eight 53-d-old rats were equally divided into 4 groups. Rats were given a purified diet (0.5% Ca and 0.35% Pi) throughout the experiment. Calcium balance and 45Ca kinetics studies using an open two-compartment model were performed over a 3-d period when rats were 60 (control), 74 , 88 and 116 d old. A simple calcium balance study, using the last group, was carried out at the age of 102 d. Compared with the control group, true intestinal calcium absorption was significantly increased at 74 d, but then decreased in a time-dependent manner up to 116 d. The apparent intestinal calcium absorption began to decrease at 88 d, and decreased further with age. An increase in urinary calcium output was observed at 74 and 88 d in phase with calcium excretion into the intestine, and both variables returned to the control level at 116 d. Calcium into bone was decreased at 74 and 116 d. Calcium from bone similarly changed with Vo+ at 74 d: however the highest value was observed at 88 d. Thereafter, Vo- reached the control level at 116 d. Calcium balance rapidly decreased at 88, 102 and 116 d, but still remained positive. Calcium intake remained steady until 102 d, but was significantly decreased at the end of the experiment. The changes in calcium flux with age can be summarized as follows: 1) bone turnover decreased, calcium excretion into the urine and feces began to increase, and true intestinal calcium absorption increased, 2) apparent intestinal calcium absorption decreased, while calcium from bone and calcium excretion into the urine and feces increased, and 3) calcium intake and calcium into bone decreased.

A clinical and histological evaluation of titanium mini-implants as anchors for orthodontic intrusion in the beagle dog.

The aim of this study was to determine the anchorage potential of the titanium mini-implant for orthodontic intrusion of the mandibular posterior teeth. Six mini-implants were surgically placed around the mandibular third premolars on each side in 3 adult male beagle dogs. On the buccal site, three mini-implants were placed distal to the apex of the distal root of the third premolar, at the interradicular septa of the third premolar, and mesial to the apex of the mesial root of the third premolar, as linearly as possible. The same procedure was performed at the lingual site on both sides of the mandibular third premolars in each dog. Bilateral interradicular mini-implants on both the buccal and the lingual sites were used as the anchorage for the intrusion of the third premolars (loaded implants) and the other mini-implants were used as control (unloaded) implants. In 6 weeks, an intrusive force (150 g) was applied between the interradicular implants on the buccal and the lingual sites by closed coil springs run across the crowns of the third premolars. After 12 to 18 weeks of orthodontic intrusion, the animals were killed and their mandibles were dissected and prepared for histologic and fluorescent observation. The results indicated that the mandibular third premolars intruded 4.5 mm, on average, after 12 to 18 weeks of orthodontic force application, with mild root resorption at the furcation area as well as the root apex. All the mini-implants remained stable during orthodontic tooth movement without any mobility or displacement. The morphometrical findings indicated that the calcification of the peri-implant bone on the loaded implants was equal to or slightly greater than those of the controls. In addition, 6 of the 36 mini-implants were removed after tooth movement, and all of them were easily removed with a screwdriver. These findings suggest that mini-implants are effective tools for the anchorage of orthodontic intrusion in beagle dogs.

Amelioration of experimental autoimmune encephalomyelitis in C57BL/6 mice by an agonist of peroxisome proliferator-activated receptor-gamma.

Peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, plays a critical role in adipocyte differentiation and glucose homeostasis. It has been implicated that PPAR-gamma functions as a regulator of cellular proliferation and inflammatory responses. In the present study, we examined whether troglitazone, a selective PPAR-gamma agonists, ameliorated experimental autoimmune encephalomyelitis (EAE) induced by administration of myelin oligodendrocyte glycoprotein (MOG) peptide 35-55 in C57BL/6 mice. We found that troglitazone attenuated the inflammation and decreased the clinical symptoms. It was suggested that the amelioration was attributed to the attenuation of pro-inflammatory cytokine gene expressions.

Endosseous titanium implants as anchors for mesiodistal tooth movement in the beagle dog.

The purpose of this study was to determine the anchorage potential of titanium implants (Branemark; 3.75 x 7 mm) with the use of a sectional arch wire technique for orthodontic mesiodistal tooth movement, as assessed by the osseointegration of implants and tooth movement. Two implants were surgically placed in healed mandibular extraction sites of the second and third premolars on each side in 4 adult male beagle dogs. The implants were surgically uncovered 18 weeks later, and second-stage abutments with soldered edgewise tubes were attached. Segmented edgewise rectangular archwires (0.017 x 0. 025 inch) with a T-loop or an L-loop were placed between the implants and the fourth premolars on both sides as the anchorage unit. One segment in each dog served as a loaded side, and the archwire was calibrated to produce 200 g of lateral force on the fourth premolar. The contralateral segment served as an unloaded side and was not subjected to orthodontic force. Sectional wires were activated biweekly 24, 28, 28, and 32 weeks, respectively, depending on the magnitude and the appearance of mesial tipping movement of the fourth premolar. After mandibular impressions were taken to measure the distance between the first molar and the fourth premolar, the animals were euthanized and dissected mandibles were prepared. The specimens were then embedded in polyester resin and cut to take backscattered electron images. On the basis of these images, the percentage of peri-implant bone volume was calculated and defined as an index of osseointegration. The differences between the initial and final fourth premolar to first molar distances varied (7.40, 8.85, 10.50, and 3.30 mm) on the loaded side, whereas the unloaded side showed no movement. Not only was there no statistical difference in the percent of peri-implant bone volume between the loaded and unloaded sides, but there was also no statistical difference between the compression and tension sides in both loaded and unloaded implants, which suggests that the implants maintained rigid osseointegration. In conclusion, the present study demonstrated that endosseous titanium implants can function as anchors for long-term orthodontic mesiodistal movement.

Significant MLR but not CTL responses against recipient antigens generated in T cells from bone marrow chimeras recovered from acute GVHD.

Lethally irradiated AKR mice received BMT from H-2D and minor lymphocyte stimulatory (Mls)-1 disparate B10.A mice. No GVHD signs were detected in AKR recipients of T cell-depleted BM cells (1 x 10(7)) alone ([B10.A --> AKR] T-). When B10.A splenic T cells (1 x 10(5)) were injected in addition to T cell-depleted BM cells ([B10.A --> AKR] T+), overt GVHD was observed. [B10.A --> AKR] T+ chimeras recovered from the GVHD 8 weeks after BMT. In T cells from these [B10.A --> AKR] T+ chimeras, a substantial population of Mls-1a-reactive Vbeta6+ T cells was present, whereas the Vbeta6+ cells were deleted in [B10.A --> AKR] T- chimeras. T cells from [B10.A --> AKR] T+ chimeras showed considerable MLR but no CTL response against AKR cells (split tolerance). Upon stimulation with AKR stimulators or anti-CD3 MoAb, T cells from [B10.A --> AKR] T+ chimeras produced significantly more IL-4 but significantly less IFN-gamma compared with those from [B10.A --> AKR] T- chimeras or unmanipulated B10.A mice. The serum level of IgG1 in [B10.A --> AKR] T+ chimeras was also significantly higher than that in [B10.A --> AKR] T- or B10.A mice. The present findings suggest that the split tolerance observed in BMT chimeras recovered from GVHD is attributable to the Th2 dominant state.

Abrogation of negative selection by GVHR induced by minor histocompatibility antigens or H-2D antigen alone.

Allogeneic bone marrow chimeras were prepared by donor and recipient combinations that differed in minor histocompatibility loci or H-2D locus alone. When 1 x 10(5) splenic T cells were inoculated in addition to T cell-depleted bone marrow cells (1 x 10(7)), clinically detectable GVHR was induced. In these GVHR chimeras, substantial numbers of T cells reactive to either donor or recipient antigens were both phenotypically and functionally detected. The mechanisms underlying the abrogation of intrathymic negative selection are discussed.

Amelioration of experimental autoimmune uveoretinitis by pretreatment with a pathogenic peptide in liposome and anti-CD40 ligand monoclonal antibody.

We have defined a peptide K2 (ADKDVVVLTSSRTGGV) that corresponds to residues 201-216 of bovine interphotoreceptor retinoid-binding protein and induces experimental autoimmune uveoretinitis (EAU)4 in H-2Ak-carrying mice (H-2Ak mice). In this study, we attempted to ameliorate EAU in the H-2Ak mice without nonspecific suppression of T cell responses. Preceding s.c. administration of liposomes including K2 (liposomal K2) specifically inhibited subsequent generation of T cell response to K2. The same result was obtained with a combination of OVA323-339 peptide and the OVA-specific TCR-transgenic T cells. It was suggested that the inhibition was mainly attributed to peripheral anergy induction of T cells specific for the peptide Ag, although specific cell death might also be involved in the inhibition. Pretreatment with liposomal K2 also considerably abolished IFN-gamma production but not IL-4 production. The specific inhibitory effect of the pretreatment with liposomal peptide was augmented by a simultaneous administration of anti-CD40 ligand (anti-CD40L) mAb. Moreover, it was shown that the pretreatment with liposomal K2 reduced both the incidence and severity of the subsequent K2-induced EAU, and the simultaneous administration of anti-CD40L mAb augmented this preventive effect by liposomal K2. Our findings demonstrate that the s.c. administration of liposomal pathogenic peptide and anti-CD40L mAb can be applied to preventing autoimmune diseases without detrimental nonspecific suppression of T cell responses.

Effective priming of cytotoxic T lymphocyte precursors by subcutaneous administration of peptide antigens in liposomes accompanied by anti-CD40 and anti-CTLA-4 antibodies.

Recently it has been shown that modulation of CD40 molecules on antigen (Ag) carrying dendritic cells (DC) can bypass T cell help, resulting in priming cytotoxic T lymphocytes (CTL) specific for the Ag. In the present study we attempted to prime peptide Ag-specific CTL by a new method in which a peptide Ag in liposome (liposomal peptide), consisting of phosphatidylserine and phosphatidylcholine (3:7), was administrated subcutaneously with anti-CD40 and/or CTLA-4 monoclonal antibodies (mAb) to mice. We found that the subcutaneous administration of the liposomal peptide with both anti-CD40 and anti-CTLA-4 mAb enhanced CTL responses comparing with those induced by the liposomal peptide alone or the liposomal peptide plus each mAb. It was shown that liposomes were critical for induction of the CTL activity. Flow cytometry analysis of a peptide-bearing DC in lymph nodes (LN) and measurement of serum IL-12 indicated that anti-CD40 mAb promoted migration of DC to the LN, where DC might differentiate and acquire ability of priming CTL. These findings provide a possibility that our procedure is applicable to cancer patients.

Fructooligosaccharide consumption enhances femoral bone volume and mineral concentrations in rats.

We examined whether the enhanced mineral absorption resulting from fructooligosaccharide (FOS) consumption affects femoral bone structure and mineral concentrations, using histomorphometrical and X-ray microanalysis. Male Wistar rats (n = 16; 42 d old) were divided into two groups, a control group (n = 8) and a FOS group (5 g/100 g FOS in the diet, n = 8). After a 3-d adaptation period, constant amounts of calcium (95 mg/d) and magnesium (8 mg/d) were fed to the rats in each group, using a pair-feeding protocol. At age 60 d, a 3-d metabolic study was initiated. Calcium and magnesium absorptions were calculated. The rats were then killed, and the right femur was embedded in polyester resin. The distal metaphysis was sagittal-sectioned, and the middle of the diaphysis and neck were cross-sectioned. Calcium, magnesium and phosphorus concentrations in the three samples were then measured. Calcium and magnesium absorptions were significantly greater in FOS-fed rats. Trabecular bone volume at the metaphysis and bone volume at the neck of the femur in FOS-fed rats were also significantly greater than those in control rats. The mineral concentration (Ca, Mg and P) in each region of the bone surface was greater in FOS-fed rats. There was a significant relationship between absorbed calcium and calcium concentrations in bone (r = 0.722, P < 0.001), and a similar relationship was found for magnesium (r = 0.720, P < 0.001). These results suggest that the enhanced calcium and magnesium absorption due to FOS consumption might enhance femoral bone volume and mineral concentrations.

Dietary fructooligosaccharides prevent a reduction of cortical and trabecular bone following total gastrectomy in rats.

Fructooligosaccharides (FOS) have been shown to stimulate the absorption of several minerals in the intestine. In the present study, the effects of FOS on osteopenia induced by total gastrectomy were examined. Twenty eight male Sprague Dawley rats were divided into 2 groups: sham-operated (SH) and gastrectomized (GX). After a one-week adaptation period following surgery, the rats were fed synthetic diets with or without 7.5% FOS for 5 weeks. The right femur was then examined by soft X-ray, and the bone mineral density (BMD) was measured. Based on the soft X-ray findings, both cancellous and cortical bone were markedly decreased in GX rats, but not in GX + FOS rats. GX rats showed a 30% lower BMD in the metaphysis and a 20% lower BMD in the diaphysis, compared with SH rats (P < 0.01). As assessed by morphometry, significant decreases were observed in cortical bone in the diaphysis and trabecular bone in the distal metaphysis (P < 0.01). On the other hand, dietary FOS completely prevented these changes following gastrectomy. These findings indicate that dietary FOS might contribute to the prevention of bone diseases following gastrectomy.

True calcium absorption in the intestine is enhanced by fructooligosaccharide feeding in rats.

Fructooligosaccharides (FOS) have been shown to stimulate apparent calcium absorption in the intestine. In this study, we examined the effect of FOS on true calcium absorption using the calcium balance in combination with the 45Ca kinetics method. Sixteen 45-d-old male Wistar rats were randomly divided into two groups, a control group (n = 8) and a FOS group (n = 8). The diet fed to the FOS group contained 5% FOS, at the expense of half of the sucrose in the control diet. After an adaptation period (3 d) and a free-access period (3 d) that were used to estimate the amount of food required for pair-feeding on the basis of calcium, all of the rats were pair-fed throughout the experiment from the age of 51 d. A constant amount of calcium was fed to the rats in each group (95 mg /d). At age 60 d, a 3-d metabolic study was started by the intravenous injection of 45Ca. Several variables were calculated on the bases of measurements of calcium intake, calcium in feces and serum, and 45Ca in feces, urine and serum. Both true and apparent calcium absorption in the intestine (Vad and Vna) and urinary calcium were significantly greater in rats that had been fed FOS. There were no differences between the groups in endogenous net calcium excretion into feces (Vf; Vad - Vna). The calcium balance was also enhanced by FOS. Calcium balance in the FOS group was significantly correlated with the absorbed calcium (r2 = 0.936, P < 0.01), as was that in the control group (r2 = 0.994, P < 0.01). These results suggest that the increased true calcium absorption and balance produced by FOS feeding might improve bone calcification.

[45Ca kinetics and calcium balance studies. A useful method for analyzing calcium movement as a whole throughout the entire body in rats].

45Ca kinetics and calcium balance studies allow for the simultaneous determination of calcium movement in the intestines, kidneys and bones. Since the flow of minerals in each of these organs depends on those in other organs, it is important to evaluate calcium movement as a whole throughout the entire body. Morphological methods can be used to evaluate local bone formation and resorption microscopically within defined windows. However, bone remodeling does not proceed equally in every part of the skeleton, and local events tend to be misread as general phenomena. Although variables such as bone formation and resorption obtained in 45Ca kinetics and calcium balance studies tell nothing about the location of these events, and the concept of compartments can be somewhat difficult to understand, this method is still useful for obtaining quite reliable results compared to those obtained by other methods. Our review describes the concept of the compartment model and explains the pertinent methods in comparison with other methods.

The application of three-dimensional quantitative computed tomography to the maxillofacial skeleton.

OBJECTIVES: To develop a method for three-dimensional quantitative computed tomography (QCT) of the maxillofacial skeleton. METHODS: The linearity of reference phantom consisting of five hydroxyapatite (HA) rods was tested under various conditions, including the size of field of view, the position of object, and reconstruction algorithm, on three different scanners. The calibrated HA concentration value and the absolute Ca content of the mandible removed from fresh cadaver was compared in order to evaluate the calibration error. RESULTS: There was a high correlation between average CT number and HA concentration in the phantom rods. There was a 4-5% difference in results from the three scanners. High density areas, corresponding to more than 75% HA, were extracted from the original image. Data were reconstructed three-dimensionally and combined using a 32-bit computer system. A three-dimensional distribution of bone density superimposed on the maxillofacial skeleton was produced. CONCLUSION: Three-dimensional QCT of the maxillofacial skeleton has potential as a means of analysing bone density. Difference in performance of different CT scanners should be considered when comparing results.

Non-phenotypic detection of osteopetrotic (op/op) mutation by using PCR-SSCP analysis.

Mice homozygous for recessive mutation osteopetrosis (op/op) on chromosome 3 provide a unique model to study the mechanism of haematopoiesis in conjunction with bone formation. Based on the DNA sequence data recently reported, we established a PCR-SSCP (polymerase chain reaction--single strand conformation polymorphism) assay which identifies an amplified fragment having an insertional point mutation present in macrophage colony-stimulating factor (M-CSF) gene of op/op mice. With this assay, three genotypes, op/op, +/op, and +/+ can be distinguished. Although heterozygous (+/op) and normal (+/+) mice could not be discriminated phenotypically, we could generate op/op mutant mice starting from a single heterozygous (+/op) mouse using only the PCR-SSCP aided screening method. This assay will permit introduction of the op mutant into any strain to generate a new animal model to study the cytokine network and haematopoiesis.

Effects of strontium on calcium metabolism in rats. II. Strontium prevents the increased rate of bone turnover in ovariectomized rats.

The effects of stable strontium were investigated in ovariectomized (OVX) rats by calcium balance and calcium kinetic studies, histomorphometric analysis and measurements of calcium levels in bone. After 10 days of pair-feeding with a control diet, 71-day-old female Wistar rats were either sham-operated (Sham group) or ovariectomized. The OVX rats were divided into two subgroups: those that were treated with strontium (OVX+Sr group, strontium intake; 87.5 mumol/day/rat) and those that were not (OVX group). Both groups were pair-fed their respective control or strontium diets for 2 weeks. Calcein and tetracycline were injected every 2 weeks from 1 week before ovariectomy to calculate the rate of bone formation in the diaphyseal femora cortex (% BFFC). In the OVX group, urinary calcium and % BFFC decreased, while bone resorption, bone formation and femora length increased at the end of the experiment, as compared with those in the Sham group. No such changes were observed in rats in the OVX+Sr group. The calcium balance, calcium levels in bone and trabecular bone volume in the metaphysis did not change in any of the three groups. These results suggest that strontium may be able to prevent the changes in bone turnover induced by estrogen deficiency.

Effects of strontium on calcium metabolism in rats. I. A distinction between the pharmacological and toxic doses.

Strontium at low doses has been used to treat osteoporosis. However, excessive doses can disturb calcium metabolism. The aim of the present study was to determine a dose that does not have any significant toxic effects on calcium contents in bone and calcium metabolism, and, consequently, to distinguish between pharmacological and toxic doses in rats. The rats were divided into a control, 0.05%-Sr, 0.10%-Sr and 0.50%-Sr groups (strontium intake approx. 0, 87.5, 175 and 875 mumol/day, respectively). All of the rats were pair-fed their respective diets containing various doses of strontium in single metabolic cages from when they were 36 to 63 days old. When the rats were 60 days old, bone formation, bone resorption, calcium balance and intestinal calcium absorption were calculated as calcium metabolic parameters over a 3-day period using calcium balance and kinetic studies. At the age of 64 days, the rats were sacrificed under anesthesia, and the femur and blood were collected. Calcium and strontium levels in the bone and serum were then measured. In the strontium groups that received less than 175 mumol/day, none of the calcium metabolic parameters were significantly affected. However, the calcium contents in the bone were significantly increased in the group that received 87.5 mumol/day group. On the other hand, in the group that received the highest dose of strontium (875 mumol/day), all of the calcium metabolic parameters measured were markedly suppressed. A decrease in calcium level in both the bone and serum was also observed.(ABSTRACT TRUNCATED AT 250 WORDS)