Biomedical subjects

K Ohya

Publications and source records attributed to K Ohya.

At least 55 records · Page 3Linked to original sources

PubMed1999

Walker 256/S carcinosarcoma causes osteoporosis-like changes through ectopical secretion of luteinizing hormone-releasing hormone.

We have shown that Walker 256/S mammary carcinoma caused osteoporosis-like changes in young female rats, accompanied by low serum estradiol and hypercalciuria without changes in the serum levels of calcium, phosphorus, and parathyroid hormone-related peptide. In this study, we investigated the cause of bone loss after Walker 256/S inoculation into female 6-week-old Wistar Imamichi rats, focusing on the sex hormone balance in the host animal. Walker 256/S-bearing rats showed characteristic osteoporosis, with a significant increase in spleen weight and a significant decrease in uterine weight by 14 days after s.c. tumor inoculation. In the in vitro bone marrow culture, mineralized nodule formation ability decreased according to the time after tumor inoculation, and tartrate-resistant acid phosphatase-positive multinucleated cell formation increased at 7 days after tumor inoculation, but it began to decrease at 14 days after tumor inoculation. This indicates that after inoculation with Walker 256/S tumor, the progenitors of osteoblasts and ostroclasts lost their balance in the bone turnover, resulting in bone resorption. On the other hand, Walker 256/S carcinoma expressed luteinizing hormone-releasing hormone (LH-RH) mRNA, and in Walker 256/S-bearing rats, the serum LH-RH level increased significantly from 3 days after tumor inoculation, whereas in the healthy control rats, this level was very low. Consequently, the serum levels of follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone were significantly lower in the tumor-bearing rats than in the healthy control rats. Because the LH-RH gene is located in the long prolactin release-inhibiting factor (PIF) gene and mRNA amplified by reverse transcription-PCR in this study contained whole LH-RH and a part of PIF, the Walker 256/S tumor is thought to express PIF. Indeed, the serum prolactin level decreased in tumor-bearing rats. The serum level of growth hormone, one of the other pituitary hormones, was not changed. Moreover, the level of an osteolytic cytokine, tumor necrosis factor alpha, increased in the serum of Walker 256/S-bearing rats, although this may be a result of the immune response of the host animal to tumor growth as well as an enlarged spleen. In conclusion, the Walker 256/S tumor lowers estrogen secretion through ectopical oversecretion of LH-RH, and then osteolytic cytokines, such as tumor necrosis factor alpha, increase in tumor-bearing rats, escape the control of estrogen, and activate osteoclasts, resulting in bone loss in a short period.

Animals↗

PubMed1999

Extracellular role of S100A4 calcium-binding protein in the periodontal ligament.

S100A4 is a member of the S100 calcium-binding protein family. S100A4 is expressed in several tissues; however, it is secreted by few cell types and its extracellular roles are unknown. In the present study we showed by in situ hybridization that periodontal ligament (PDL) cells express the S100A4 mRNA. Immunolocalization of the S100A4 protein in cryosections of PDL and analyses of PDL cell culture medium revealed that PDL cells secrete the S100A4 protein both in vivo and in vitro. Interestingly, addition of a recombinant mouse S100A4 protein to a bone marrow cell culture inhibited mineralized nodule formation in a concentration-dependent manner. This is the first report of an extracellular role for S100A4 as an inhibitor of mineralization. The PDL space is kept free of mineralization and S100A4 may be one of the factors responsible for such phenomenon.

Animals↗

PubMed1999

Desmoglein 1 and desmoglein 3 are the target autoantigens in herpetiform pemphigus.

OBJECTIVE: To determine the cell surface autoimmune target of herpetiform pemphigus (HP). DESIGN: Serum samples of HP were examined by immunoblot studies with human epidermal extracts, enzyme-linked immunosorbent assay with baculovirus-expressed recombinant desmoglein (rDsg) 1 and rDsg3, and immunoadsorption assay with rDsg. PATIENTS: Twenty serum samples were obtained from patients with HP who have typical clinical and histological features. All serum samples showed positive staining against keratinocyte cell surfaces by indirect immunofluorescence studies with healthy human skin. RESULTS: Immunoblot results showed that of 17 HP serum samples, only 5 reacted with a 160-kd band and 1 reacted with a 130-kd band. Results of enzyme-linked immunosorbent assays with rDsg1 and rDsg3 demonstrated that of 20 HP serum samples, 16 were positive against Dsg1 and 4 were positive against Dsg3. No serum samples reacted with both. Furthermore, in 19 of 20 HP serum samples, immunoreactivity against keratinocyte cell surfaces was completely removed by preincubation with rDsg1 and rDsg3 as shown by indirect immunofluorescence, excluding a possibility that these HP sera contain autoantibodies against other cell surface molecules. CONCLUSIONS: Dsg1 and Dsg3 are the major cell surface target molecules of HP, suggesting that most cases of HP are clinical variants of pemphigus foliaceus and that the rest might be variants of pemphigus vulgaris.

Autoantigens↗

PubMed1999

Charcot-Marie-Tooth disease type 1A duplication by PCR analysis.

By generating new junctional fragments from the recombinant Charcot-Marie-Tooth (CMT) 1A-REPs in CMT1A patients, a 3.2-kb recombination hot spot is observed in three quarters of CMT1A patients. By a polymerase chain reaction (PCR) method the authors analyzed eight patients CMT1A duplication, confirmed by Southern blot, to detect a recombination hot spot. Four patients had a novel 3.2-kb junctional fragment by PCR analysis. These four patients with a novel 3.2-kb junctional fragment had an abnormal 1789-bp fragment in addition to 1986-bp fragment after NsiI digestion (type 1). One patient who demonstrated no novel 3.2-kb junctional fragment had an abnormal 336-bp fragment in addition to 265 bp (type 2). Three patients with CMT1A duplication were not diagnosed as having CMT1A on the basis of PCR analysis. The PCR-based DNA test is valuable for screening to detect CMT1A duplication.

Adolescent↗

PubMed1999

Anabolic effect of aminoterminally truncated fibroblast growth factor 4 (FGF4) on bone.

Fibroblast growth factor 4 (FGF4), a member of the FGF family, plays several important roles in bone development during embryogenesis. Systemic administration of FGF4 increases bone mass in rats, which suggests the potential therapeutic usefulness of this growth factor in treatment for osteopenia and in bone regeneration. We investigated the length of FGF4 required to exert its anabolic effects, because this information may be useful in developing new molecules to mimic the effects of FGF4. Because the active site of FGF family molecules is in the carboxylterminal region, we produced aminoterminally truncated recombinant human FGF4s (rhFGF4s) of different sizes. Human FGF4 cDNA containing almost the full length of the coding region (573 bp, 191 amino acid residues) was inserted into pUC18 vector and then deleted from the 5' end using the ExoIII system. Each of the deleted FGF4 cDNAs was subcloned into a pET29(+) expression vector. Differently sized recombinant proteins were expressed in the BL21(DE3)pLysS Escherichia coli strain and then purified. The growth-stimulative effects on NIH3T3 cells of each recombinant protein were examined by means of MTT colorimetric assay. Full-length and the shortened recombinant proteins, which stimulated NIH3T3 cell growth, were then subcutaneously administered into male ddY mice (6 weeks old) every day for 2 weeks. Bone mineral density (BMD) was measured using dual-energy X-ray absorptiometry (DEXA) and peripheral quantitative computed tomography (pQCT). The rhFGF4 of 134 amino acid residues, the region homologous to other members of the FGF family, exerted a growth-stimulative effect on NIH3T3 cells comparable to the full-length version of FGF4; however, the shortest version, with 111 amino acid residues, showed a limited growth-stimulative effect. Systemic administration of the rhFGF4 of 134 amino acid residues increased the bone mineral density (BMD) of femurs at a dose of 0.1 mg/kg, which was comparable to that of the full-length rhFGF4. DEXA analysis, pQCT analysis, soft X-ray photos, and contact microradiographs revealed an increase in femoral trabecular bone in FGF4-treated animals; an increase in bone formation was also evident upon histomorphometric analysis. These results indicate that the region of FGF4 that is homologous to other FGF family members provides a sufficient anabolic effect in bone and that this recombinant protein is potentially useful as a therapeutic agent in bone.

3T3 Cells↗

PubMed1999

Trabecular bone turnover, bone marrow cell development, and gene expression of bone matrix proteins after low calcium feeding in rats.

Low-calcium-fed animals have been accepted as one of the experimental models showing a reduction in bone mass. However, the effects of short-term low-calcium feeding on bone turnover, the development of osteoprogenitor cells, and gene expression of bone matrix proteins have not been reported. In this study, we examined the effect of a low-calcium diet on rat tibia and analyzed the changes in the bone by histomorphometry, bone marrow cell culture, and in situ and Northern hybridization of the bone matrix proteins. Rats were fed either a low-calcium diet (0.05% Ca) or a normal calcium diet (0.5% Ca) using the pair feeding technique. They were killed at day 0, 12 h, and days 1, 2, and 3. In the low-calcium group, the serum parathyroid hormone (PTH) level was temporarily increased in 12 h after feeding the low-calcium diet. Bone mineral density in the trabecular bone was significantly decreased from 1 day after the low-calcium feeding, but cortical bone did not show any changes during the experimental period. The bone volume per tissue volume in the proximal tibia also decreased from day 1 in the low-calcium group. The number of osteoclasts and osteoblasts on the trabecular bone surface was increased in the low-calcium group compared with the normal-calcium group. An ex vivo study showed that the number of progenitors of osteoclasts and osteoblasts in bone marrow was also increased in the low-calcium group of rats. The localization of type I collagen mRNA was observed in osteoblasts in the low-calcium group. The Northern hybridization study showed that the gene expression of type I collagen, osteopontin, and osteocalcin was increased at day 3 in the low-calcium group. These results indicated that the trabecular bone surface quickly responded to the low-calcium feeding and that bone remodeling activity was activated probably by PTH. The changes in bone marrow cell populations and the gene expression of bone matrix proteins are closely associated with increased bone turnover induced by the low-calcium diet, resulting in rapid bone loss of the trabecular bone.

Animals↗

PubMed1999

Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus.

Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Patients with pemphigus have circulating antidesmoglein (Dsg)1 and/or anti-Dsg3 IgG autoantibodies. We have previously developed enzyme-linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 expressed by baculovirus as a diagnostic tool for pemphigus. The purpose of this study was to evaluate the practical application of these ELISAs for clinical use with a large number of serum samples. We used 81 PV sera, 48 PF sera, 114 bullous pemphigoid (BP) sera, 124 collagen disease sera, nine sera of other non-pemphigus bullous diseases and 179 normal control sera. A cut-off value was determined by receiver-operating-characteristic plots. Forty-seven of 48 PF sera (97.9%) were positive in the Dsg1 ELISA and 79 of 81 PV sera (97.5%) were positive in the Dsg3 ELISA, while only two (1. 1%) and four (2.2%) of 179 normal sera were positive in Dsg1 and Dsg3 ELISAs, respectively. However, some disease control sera of BP and collagen diseases exceeded the cut-off value. Introduction of a grey zone helped to decrease the number of these false-positive sera. Furthermore, in three patients studied, the respective Dsg1 and Dsg3 ELISA scores showed parallel fluctuation with the disease activity along the time course. We conclude that Dsg1 and Dsg3 ELISAs provide a simple, sensitive and highly specific assay for the diagnosis of patients with PV and PF and that these ELISAs may be a valuable tool to monitor the disease activity. We also propose diagnostic criteria for pemphigus based on ELISA reactivity: if a serum is positive against Dsg3 it indicates a diagnosis of PV, regardless of reactivity against Dsg1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.

Aged↗

PubMed1999

In situ hybridisation study of type I, II, X collagens and aggrecan mRNas in the developing condylar cartilage of fetal mouse mandible.

The aim of this study was to investigate the developmental characteristics of the mandibular condyle in sequential phases at the gene level using in situ hybridisation. At d 14.5 of gestation, although no expression of type II collagen mRNA was observed, aggrecan mRNA was detected with type I collagen mRNA in the posterior region of the mesenchymal cell aggregation continuous with the ossifying mandibular bone anlage prior to chondrogenesis. At d 15.0 of gestation, the first cartilaginous tissue appeared at the posterior edge of the ossifying mandibular bone anlage. The primarily formed chondrocytes in the cartilage matrix had already shown the appearance of hypertrophy and expressed types I, II and X collagens and aggrecan mRNAs simultaneously. At d 16.0 of gestation, the condylar cartilage increased in size due to accumulation of hypertrophic chondrocytes characterised by the expression of type X collagen mRNA, whereas the expression of type I collagen mRNA had been reduced in the hypertrophic chondrocytes and was confined to the periosteal osteogenic cells surrounding the cartilaginous tissue. At d 18.0 of gestation before birth, cartilage-characteristic gene expression had been reduced in the chondrocytes of the lower half of the hypertrophic cell layer. The present findings demonstrate that the initial chondrogenesis for the mandibular condyle starts continuous with the posterior edge of the mandibular periosteum and that chondroprogenitor cells for the condylar cartilage rapidly differentiate into hypertrophic chondrocytes. Further, it is indicated that sequential rapid changes and reductions of each mRNA might be closely related to the construction of the temporal mandibular ramus in the fetal stage.

Aggrecans↗

PubMed1999

Expression of the myotonic dystrophy locus-associated homeodomain protein in congenital myotonic dystrophy.

DM locus-associated homeodomain protein (DMAHP), a newly recognized homeodomain protein, is a candidate flanking gene and expressed in various tissues. We examined the expression of the DMAHP in tissues from a congenital myotonic dystrophy patient in comparison to that of control tissues by using semiquantitative reverse transcriptase-polymerase chain reaction. Reduced expression of DMAHP was observed in tissues from the patient with congenital myotonic dystrophy. This finding supports that reduced expression of DMAHP is related to the CTG repeat expansion.

Female↗

PubMed1999

Effects of XT-44, a phosphodiesterase 4 inhibitor, in osteoblastgenesis and osteoclastgenesis in culture and its therapeutic effects in rat osteopenia models.

We have reported that denbufylline, a phosphodiesterase 4 (PDE4) inhibitor, inhibits bone loss in Walker256/S tumor-bearing rats, suggesting therapeutic potentiality of a PDE4 inhibitor in osteopenia. In the present study, effects of a new PDE4 inhibitor, 1-n-butyl-3-n-propylxanthine (XT-44), in bone were evaluated in cell cultures and animal experiments. In rat bone marrow culture, XT-44 stimulated mineralized-nodule formation, whereas it inhibited osteoclast-like cell formation in mouse bone marrow culture. In Walker256/S-bearing rats (6-week-old female Wistar Imamichi rats), rapid decrease in bone mineral density (BMD) was prominent, and oral administration of XT-44 (0.3 mg/kg, every 2 days) inhibited the decrease in BMD. In the second animal experiment, female Wistar rats (6-week-old) were sciatic neurectomized, and XT-44 was orally administered to these rats every 2 days for 4 weeks. XT-44 administration (0.3 mg/kg) recovered BMD in these neurectomized animals. Furthermore, 19-week-old, female Wistar rats were ovariectomized (OVX), and 15 weeks after surgery, these rats were orally administered XT-44 every 2 days for 8 weeks. XT-44 treatment (1 mg/kg) increased the BMD of OVX rats. These results indicate that XT-44 could be a candidate as a therapeutic drug for treating osteopenia including osteoporosis.

1-Methyl-3-isobutylxanthine↗

PubMed1999

Postmenopause-like bone loss by mammary carcinoma Walker256/S which secretes luteinizing hormone-releasing hormone.

When Walker 256/S carcinosarcoma (W256/S) was subcutaneously inoculated into the back of mature female Wistar Imamichi rats (10-week-old), the tumor grew rapidly and caused increases in the urinary excretions of calcium and hydroxyproline, without changes in the serum concentrations of calcium and inorganic phosphorus. Furthermore, osteoporosis-like changes in the femurs and decrease in uterus weight were observed, as previously reported for W256/S-bearing young rats. In the healthy mature female rats, the estrus cycle passed through four stages (proestrus, estrus, metestrus and diestrus) within 4 to 5 days, with a peak of serum estradiol and progesterone levels in the proestrus stage. On the other hand, after subcutaneous inoculation of W256/S into the rats, the estrus cycle tended to pause upon the metestrus or diestrus stage, accompanied with significantly low estradiol and progesterone levels in serum. W256/S tumor produced and secreted luteinizing hormone-releasing hormone (LH-RH). In conclusion, it seems that the ectopical secretion of LH-RH from the tumor resulted in the decrease in the secretion of gonadotropic hormones, following low level of sex hormones and stopping the estrus cycle. Therefore, W256/S-bearing rats may be a model for osteoporosis of hypoovarianism or postmenopause.

Alkaline Phosphatase↗

PubMed1999

Intermittent inhibition of dentin mineralization of rat incisors under continual infusion of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) using a subcutaneous mini osmotic pump.

The inhibitory effect of the continual administration of 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) (8 mgP/kg/day) through a mini osmotic pump on dentin mineralization was examined in relation to the diurnal rhythm of the rat and compared with that of daily injections of same amounts of HEBP known to inhibit dentin mineralization. After daily injections of HEBP, a series of alternating rows of mineralized and non-mineralized dentin islands appeared in the newly formed portion of the crown-analogue of rat incisors. A similar phenomenon occurred under the continual administration of HEBP in rats raised either under regular environmental photofraction or constant lighting conditions. The average distance between the adjacent mineralized dentin islands was 521.0 +/- 51.3 microns in the injected rats. After continual HEBP administration, this was 426.0 +/- 13.2 microns and 416.5 +/- 19.4 microns under ordinary photofraction and constant light, respectively. Although the pattern of individual mineralized dentin islands tended to become irregular in nocturnal rats, no statistical difference was noted between the two values. Rows of mineralized and non-mineralized dentin islands also appeared in the root analogue dentin. No sign of the intermittent inhibition of mineralization was recognized in mesodermal hard tissues other than dentin in the HEBP-affected animals. These data implicate the presence of intrinsic cycles in dentin mineralization at the growing end of rat incisors independent of environmental photofraction as well as the ameloblast function.

Animals↗

PubMed1999

Detection of anti-LKM-1(anti-CYP2D6) by an enzyme-linked immunosorbent assay in adult patients with chronic liver diseases.

Anti-liver kidney microsome-1 (LKM-1) autoantibody, which is a serological marker for autoimmune hepatitis type II, recognizes Cytochrome P450 IID6 (CYP2D6). This autoantibody is also detected in a portion of patients with chronic hepatitis C. Anti-LKM-1 has been measured by indirect immunofluorescence (IF) using rat liver and kidney sections. However, this method has some problems in specificity and is so laborious to handle with many samples. In this study, in order to determine anti-LKM-1, we established an enzyme-linked immunosorbent assay (ELISA) for anti-CYP2D6 using a recombinant CYP2D6 fusion protein. We studied sera from 29 patients positive for anti-LKM-1 by the new ELISA. We further studied sera from a total of 301 patients with various liver diseases and 100 sera from normal controls negative for anti-LKM-1 by the new ELISA. The specificity of the ELISA was ascertained by absorption tests using sera positive for anti-LKM-1. In 29 sera from patients positive for anti-LKM-1 by IF, we found a good correlation between the logarithms of the antibody titers determined by IF and ELISA indexes obtained by our new method. Anti-CYP2D6 was positive in 12 of 12 (100%) patient with autoimmune hepatitis type II and 16 of 17(94.1%) with chronic hepatitis C positive for anti-LKM-1 by IF. In other 401 sera negative for anti-LKM-1 by IF, anti-CYP2D6 was all negative except a few sera. We established a new ELISA for anti-LKM-1 (anti-CYP2D6). This ELISA system is sensitive, antigen-specific and easy to be done. Therefore, this assay allows a routine test of many serum samples, especially for diagnosing autoimmune hepatitis type II.

Adult↗

PubMed1999

[3D-CT cystography of submucosal elevated lesion of the bladder: report of two cases].

We report 2 cases of rare inflammatory disease of the bladder arising from the bladder submucosa: eosinophilic cystitis in a 33-year-old woman and inflammatory pseudotumor of the bladder in a 41-year-old man. 3D-CT cystography demonstrated submucosal tumorous lesions clearly and enabled the evaluation of mucosae of lesions especially showed the bridging fold-like appearance of the submucosal tumorous lesion in eosinophilic cystitis.

Adult↗

PubMed1999

Effects of mechanical stress on the mRNA expression of S100A4 and cytoskeletal components by periodontal ligament cells.

The periodontal ligament (PDL) functions under constant mechanical stress, and PDL cells obviously control PDL functions under such conditions. We have previously found that the mRNA expression of the Ca2+-binding protein S100A4 and beta-actin is higher in the PDL from erupted teeth than in the PDL from teeth under eruption. This suggested a role for S100A4 in the response of PDL cells to mechanical stress, possibly by coupling Ca2+ and the cytoskeletal system. In the present study, we investigated the direct effects of cyclical stretching on the mRNA expression of S100A4 and two cytoskeletal components (beta-actin and alpha-tubulin) by PDL cells. In Northern blotting analysis, the expression of S100A4, beta-actin, and alpha-tubulin mRNAs was higher in the PDL from fully erupted and functional bovine teeth than in partially erupted ones. Similarly, when bovine PDL cells were mechanically stimulated by means of the Flexercell Strain Unit, the expression of S100A4, beta-actin, and alpha-tubulin mRNAs increased over the control levels. The results of our present study indicate that S100A4 is involved in the responses of PDL cells to mechanical stress possibly by coupling Ca2+ to the cytoskeletal system in these cells.

Actins↗

PubMed1998

Behavior of bone marrow cells cultured on three different coatings of gel-derived bioactive glass-ceramics at early stages of cell differentiation.

Previous studies have shown different macrophage responses to three compact pellets (with slightly different chemical composition) of gel-derived bioactive glass-ceramics of the CaO-P2O5-SiO2 system. In the present study primary bone marrow cells directed in vitro to form osteoblastic or osteoclastic cells were cultivated on glass slides coated by these three glass-ceramics. Glass slides were used as controls. In osteoblastic cultures alkaline phosphatase activity varied, depending on the type of coatings. Northern analysis showed high mRNA expressions of bone-related proteins on both the glass-ceramics and control glass. Mineralized nodules were not formed on the control glass, but coating glass slides with the glass-ceramics promoted mineralization without any substantial differences between the types of coatings. In osteoclastic cultures, the normal morphology of tartrate resistant acid phosphatase-positive multinucleated cells on standard culture plates was altered on the control glass and the glass-ceramics. The number of these cells differed, depending on the type of coatings, with no particular differences in the arrangement of F-actin by these cells. These analyses demonstrate complete biocompatibility of the glass-ceramic coatings but not the control glass, on which the cells failed to form mineralized nodules. The phenotype expression of the cells appeared to be influenced by microstructure, surface roughness, and the general character of the coatings rather than their surface reactivity.

Animals↗

PubMed1998

A small direct tandem duplication of the myelin protein zero gene in a patient with Dejerine-Sottas disease phenotype.

We present a male patient with Dejerine-Sottas disease phenotype, who had a small direct tandem duplication of the Po gene. The pathology of the sural nerve showed hypomyelinated fibers with absence of active demyelination and onion-bulb formations composed of two parallel layers of basement membrane, consistent with congenital hypomyelination neuropathy (CHN). However, his clinical features were more severe than those of previously reported CHN patients. A GGCA insertion was identified at the position of nucleotide 560 in the myelin protein zero (Po) gene. This insertional mutation was located in exon 4 coding for the transmembrane domain of the Po gene and caused a shift of reading frame, creating a stop codon. The mutation of the transmembrane domain probably has the largest impact on Po function. The mutation was not identified in both parents.

Child↗

PubMed1998

Confocal laser scanning-microscopic observations on the three-dimensional distribution of oxytalan fibres in mouse periodontal ligament.

The purpose was to improve confocal laser scanning-microscopic (CLSM) techniques to observe the three-dimensional (3-D) distribution of oxytalan fibres in mouse periodontal ligament and to clarify the 3-D relation between those fibres and blood vessels. As aldehyde fuchsin is a contrast agent and the specific wavelength affects the depth of penetration, CLSM reflectance imaging of oxytalan stained with aldehyde fuchsin after oxidization provides strong contrast. Oxytalan fibres, whose precise roles have not yet been clarified, are connective tissue fibres present in human periodontal ligament in addition to collagen fibres. Despite many studies on their arrangement and biomechanical characteristics, their 3-D distribution in relation to other structures has never been reported. Mandibular first molars of mice were sectioned mesiodistally, pre-oxidized by monopersulphate compound (Oxone), stained with aldehyde fuchsin and examined by CLSM. CLSM images clearly revealed the 3-D distribution, and relation of oxytalan fibres to blood vessels and other structures, as well as their branching patterns in the periodontal ligament. The marked anatomical correlations between the direction, distribution and branching patterns of oxytalan fibres and blood vessels suggest that they interact to perform a specialized physiological role in the periodontal ligament.

Animals↗