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Biomedical subjects

M Kagayama

Publications and source records attributed to M Kagayama.

At least 37 records · Page 2Linked to original sources

Implantation of octacalcium phosphate (OCP) in rat skull defects enhances bone repair.

Synthetic octacalcium phosphate (OCP) enhances bone formation if implanted into the subperiosteal region of murine bone. Such implanted OCP may be resorbed and replaced by bone with time. We hypothesized that OCP could be used as an effective bone substitute. To test this hypothesis, we designed the present study to investigate if bone repair in a rat skull defect is enhanced by the implantation of OCP. Rats were divided into two groups: OCP-treated animals and untreated controls. Six rats from each group were fixed at 4, 12, and 24 weeks after implantation. A full-thickness standardized trephine defect was made in the parietal bone, and synthetic OCP was implanted into the defect. After being examined radiographically, the specimens were decalcified and processed for histology. OCP implantation significantly promoted bone repair compared with the controls. A statistical analysis showed an increase in the area of radiopacity within the skull defect between week 4 and week 12. Histologically, bone was formed on the implanted OCP and along the defect margin at week 4. At week 12, the implanted OCP was surrounded by newly formed bone. At week 24, the defect was almost completely filled with bone. In the control, bone formation was observed only along the defect margin. The present results demonstrate that OCP could be used as an effective bone substitute.

Animals↗

Type X collagen is not localized in hypertrophic or calcified cartilage in the developing rat trachea.

Our previous studies have shown that rat tracheal chondrocytes become larger and hypertrophic, and that the cartilage matrix calcifies during development. Type X collagen is a short collagen molecule identified in hypertrophic and calcified cartilage in the growth plate of long bones during endochondral ossification. The present study was designed to investigate the distribution of type X collagen in rat tracheal cartilage during development before and after hypertrophization and calcification. Tracheas from postnatal Wistar rats, newborn, and at 4, 8 and 10 weeks were fixed along with hind limbs from newborn rats. Serial sections were made and adjacent sections were processed for von Kossa staining or immunohistochemistry for type X collagen. In addition, the immunoreactivity to type II collagen was examined as a control. The anti-type X collagen antibody stained hypertrophic and/or calcified cartilage in the newborn rat tibia. The immunoreaction for type X collagen was localized in the uncalcified peripheral region of tracheal cartilage in 4, 8 and 10-week-old rats. In contrast, the anti-type X collagen antibody did not show immunoreactivity to hypertrophic or calcified cartilage in the central region of the 10-week-old rat tracheal cartilage. The present study has suggested that type X collagen is not involved in hypertrophization of chondrocytes or calcification of the matrix in developing rat tracheal cartilage.

Animals↗

An immunohistochemical study of the localization of biglycan, decorin and large chondroitin-sulphate proteoglycan in adult rat temporomandibular joint disc.

To analyse regional variations in extracellular matrix components of adult rat temporomandibular joint discs, immunohistochemical techniques were used to examine the localization of two small dermatan-sulphate proteoglycans, biglycan and decorin, and a large chondroitin-sulphate proteoglycan. Staining for biglycan was intense in the posterior band, although it had a rather weak and even distribution throughout the disc. In contrast, staining for decorin was faint in the intermediate zone and the central part of the posterior band, moderate in the anterior and posterior attachments and most intense in the junction between the anterior band and attachment. The upper surface of the disc stained more intensely than the lower. Similarly, there was intense staining for large chondroitin-sulphate proteoglycan in the peripheral band, but both the anterior and the temporal parts of the posterior attachments were faintly stained. These results demonstrate marked regional differences in the expression of biglycan, decorin and large chondroitin-sulphate proteoglycan in the temporomandibular joint discs of adult rats. These variations probably reflect the different biomechanical environments caused by the complicated articulatory functions of the temporomandibular joint.

Animals↗

Epithelial rests colocalize with cementoblasts forming acellular cementum but not with cementoblasts forming cellular cementum.

Epithelial rests of Malassez and cementoblasts were examined in the rat molars during the early stages of root formation using an antilaminin antibody and/or peanut agglutinin (PNA), and an antiosteocalcin (OC) antibody, respectively. The roots of the first molars were used for study. The antilaminin antibody stained the basement membrane surrounding the epithelial root sheath and epithelial rests. The basement membrane of the epithelial root sheath was continuous, but that of the epithelial rests was discontinuous. The cells of epithelial rests and epithelial root sheath were positive for PNA. The structural characteristics of the epithelial rests were seen in the sections stained doubly with PNA and the antilaminin antibody. The cells of epithelial rests were fibroblast-like and formed a fine mesh in 2-week-old rats. In 3-week-old rats, the epithelial rests were also present at the coronal half of root surface, showing typical cell cords, but were not present at the apical part of the root surface where the cellular cementum covered the root dentin. At the root apex of 3-week-old rats, the cells of epithelial rests forming fine meshes were seen near the epithelial root sheath. The anti-OC antibody stained cementoblasts lining acellular and cellular cementum. The sections doubly stained with the anti-OC and the antilaminin antibodies or PNA further revealed the close relation between epithelial rests and cementoblasts. The OC-positive cells lining acellular cementum or dentin were localized very close to the epithelial rests. In contrast, the OC-positive cells lining cellular cementum did not show close association with the epithelial cells, except the cells located most apically where the basement membrane of the epithelial root sheath is disrupted and the initial cellular cementum begins to be formed. The present results suggest that the epithelial rests and/or the discontinuous basement membrane of them may have a role for the acellular cementum formation at least in the early stage of root formation.

Animals↗

BMPs induce endochondral ossification in rats when implanted ectopically within a carrier made of fibrous glass membrane.

BACKGROUND: Bone morphogenetic proteins (BMPs) replicate the process of embryonic bone formation when implanted in ectopic sites. Our previous studies have indicated that BMPs can induce intramembranous ossification, i.e., direct bone formation without preexisting cartilage when implanted in rats subcutaneously by using the fibrous collagen membrane (FCM) as a carrier for implanting BMPs (Sasano et al. 1993. Anat. Rec., 236:373-380). The present study was designed to investigate how the physicochemical property of the carrier material influences the process of bone formation induced by BMPs, using a carrier made of fibrous glass membrane (FGM). METHODS: BMPs, partially purified from bovine metatarsal bones, were added to an FGM carrier and implanted subcutaneously in rats. The implants were analyzed at weekly intervals, and the osteogenic process induced by BMPs was examined by histology and immunohistochemistry for cartilage and bone formation. RESULTS: Neither cartilage nor bone were observed after week 1. Cartilage formation occurred within the carrier after week 2, although no bone formation was seen. The cartilage matrix showed immunoreactivity for types II, X, and I collagen. Bone was induced on the previously formed cartilage after week 3. The bone matrix stained with anti-osteocalcin antibody and with anti-type I collagen antibody. The cartilage was replaced by bone and bone marrow after week 10. CONCLUSIONS: BMPs cause endochondral ossification when administered with an FGM carrier. The physicochemical property of the carrier may be involved in the BMP-induced phenotype expression of bone and cartilage.

Animals↗

Immunohistochemical evidence for the existence of rat cytosolic sialidase in rat skeletal muscles.

Histochemical evidence is required to demonstrate the presence of biochemically defined cytosolic sialidase. To meet this requirement, we examined the immunohistochemical localization of the enzyme in rat skeletal muscles. Sections of chemically fixed tissues were incubated with a polyclonal antibody raised against a synthetic peptide which corresponded to a part of the enzyme protein. After incubation with the primary antibody, cryosections for fluorescence microscopy and resin sections for electron microscopy were incubated with a fluorochrome- and colloidal gold-labeled secondary antibody, respectively. Immunofluorescence was diffusely distributed in the muscle fibers and was also found in the perimysium and blood vessels. Many immunogold particles were scattered over the sarcoplasm, myofibrils, nucleoplasm, and matrix of mitochondria. The immunogold particles were also found in the equivalent compartments of axons, Schwann cells, and cells of endomysium and blood vessels. The specificity of the primary antibody was elucidated by immunoblotting and an immunoprecipitation test. These findings clearly indicate that this type of sialidase is essentially located in the cytosolic compartment. Consequently, the name, cytosolic sialidase, will be appropriate for this enzyme. Additionally it is indicated that this enzyme is also present in cells other than skeletal muscle fibers.

Animals↗

Localization of types I, II and III collagen and glycosaminoglycans in the mandibular condyle of growing monkeys: an immunohistochemical study.

In order to analyse the regional and age-related variations of primate condyles, immunohistochemical techniques were used to examine the localization of types I, II and III collagen and a variety of glycosaminoglycans in distinct anteroposterior regions of the mandibular condyle of two growing female rhesus monkeys (Macaca mulatta). In the juvenile monkey staining for types I and III collagen was weak in the fibrous tissue layer, intense in the pre-cartilaginous tissue layer and faint in the cartilaginous tissue layer; staining was significantly more intense in the posterosuperior and posterior regions than in the anterior region. Similarly, staining for cartilage-characteristic extracellular matrices, including type II collagen and keratan sulfate, was intense in the cartilaginous tissue layer of the posterior condyle. In contrast, in the late-adolescent monkey staining for the extracellular matrices was more intense in the anterior half of the condyle (i.e. from the anterior to the posterosuperior region) than in the posterior region, and most intense in the posterosuperior region. The results demonstrate that marked regional differences exist in the phenotypic expression of the extracellular matrices in the mandibular condyles of growing monkeys and that these differences vary between different developmental stages. The variations probably reflect the predominance of competing growth and articulatory functions in the mandibular condyles.

Aging↗

Confocal microscopy of cementocytes and their lacunae and canaliculi in rat molars.

The present study was designed to analyze the morphological characteristics of cementocytes and osteocytes. The maxillae of 10-week-old Wistar rats were used for observations. Non-decalcified ground sections stained vitally with fluorescence dyes and decalcified frozen sections stained with FITC-phalloidin were examined by confocal microscopy. Calcein and alizarin red stained the calcification front of bone, cementum, and dentin intensely. In addition, lacunae and canaliculi of cementocytes and osteocytes as well as dentinal canals were stained with the fluorescent dyes. The staining of lacunae and canaliculi was less intense than that of the calcification front of bone, cementum and dentin. The canaliculi of cementocytes and osteocytes were connected with the canaliculi extending from the calcification front of cementum and bone, respectively. The canalicular density was less in the cellular cementum than in the bone. Areas devoid of canaliculi were numerous in the cellular cementum, whereas areas devoid of canaliculi were scarce in the alveolar bone. Further, the lacunae of cementocytes showed various shapes, from oval to tubular, while the lacunae of osteocytes were invariably oval. The cell body and the cytoplasmic processes of cementocytes were positive for FITC-phalloidin within the extracellular matrix of cellular cementum, which was negative. The distribution of actin filaments in the osteocytes and the cementocytes was predominantly cortical and appeared to be closely associated with the cell membrane of the cell bodies and the cytoplasmic processes. Intense staining was seen at the proximal part of the cytoplasmic processes in both osteocytes and cementocytes, showing a punctuated structure of the cells that was more frequent in osteocytes than in cementocytes. The stress fiber known to be present in most of the cultured cells was not evident in the these cells in situ. The cells incorporated in the cementodentinal junction were strongly stained with FITC-phalloidin. The distribution pattern of the cytoplasmic processes stained with FITC-phalloidin was similar to that of the canaliculli stained vitally. The cytoplasmic processes of osteocytes and cementocytes were connected with those of cells lining the surface of bone and cementum. The present result-that lacunae and canaliculi of cementocytes were stained vitally with the fluorescence dyes-suggests that cementocytes may have a role in secondary calcification of cellular cementum. Further, the lower density of cytoplasmic processes in cementocytes than in osteocytes suggests a lack of complexity in the intercellular network within the cellular cementum.

Actins↗

Localization of types I, II and X collagen and osteocalcin in intramembranous, endochondral and chondroid bone of rats.

Chondroid bone is a unique calcified tissue intermediate between bone and cartilage. To clarify its characteristics, we examined the distributions of the ECMs associated with chondrogenic differentiation and matrix calcification in the chondroid bone of the rat glenoid fossa, and compared them to those in two typical bone tissues, alveolar bone of the maxilla (intramembranous bone) and the growth plate of long bone (endochrondral bone), using immunofluorescence techniques. Morphologically, the glenoid fossa consisted of the fibrous, progenitor and cartilaginous cell layers and the cartilaginous cell layer was further divided into the superficial non-hypertrophic layers (secondary cartilage) and the deep hypertrophic cell layers (chondroid bone). The co-distribution of type I and type II collagens was observed in secondary cartilage and chondroid bone, whereas type X collagen was restricted to the pericellular matrix of hypertrophied cells (chondroid bone). Osteocalcin, which was absent from the calcified cartilage of endochondral bone formation, was also present in the ECM of the chondroid bone, but not in cells. These results demonstrate that chondroid bone of rats, which is adjacent to secondary-type cartilage in the glenoid fossa, has phenotypic expressions associated with both hypertrophied chondrocytes and osteocytes.

Animals↗

Development of interglobular dentine in rat molars and its relation to maturation of enamel.

The development of interglobular dentine in the first upper and lower molars of Wistar rats aged 3, 7, 14, 21, 42 days was examined histochemically using a lectin, succinyl wheat germ agglutinin (sWGA), which is specific for N-acetyl-D-glucosamine. sWGA stained the interglobular dentine, predentin and Golgi area of odontoblasts. Interglobular dentine was not formed in the first molars of 3-day rats, but appeared in those of 7-day rats near the enamel-free area. In 14-day rats, interglobular dentine was present in most areas of the coronal dentine except the cervical area. At the interface between dentine and predentin, numerous sWGA-negative calcospherites were seen, suggesting that the interglobular dentine is formed actively there. In 21-day rats, the interlobular dentine was more numerous than in 14-day rats. Interglobular dentine was present in the cervical root dentine as well as in the coronal dentine, including the cervical area. The distribution of interglobular dentine in 42-day rats was similar to that in 21-day rats, but fluorescence of sWGA binding was less intense in the former. Because the development of interglobular dentine appeared to be time and position specific its relation to the stages of ameloblasts was analysed. Thin enamel matrix was formed at cusps in molars of 3-day rats and thickness of enamel matrix increased in 7-day rats. In these teeth, the ameloblasts were at the differentiating or secretory stage. The Golgi area and Tomes' processes of the secretory ameloblasts, the cells of intermediate layer and the enamel matrix were weakly positive with sWGA. The epithelial cells at the enamel-free area were also stained with sWGA. In 14-day rats, most of the ameloblasts in the first maxillary molars were at the maturative stage except in the cervical area, where the ameloblasts were at the transitional stage. sWGA stained the distal border and the Golgi area of the maturative ameloblasts as well as the cells of the papillary layer. The distal border of the maturative ameloblasts appeared either thick or thin, suggesting a ruffle-end and smooth-end of the cells. Ameloblasts were absent in the first molars of 21-day rats and the cervical part of the enamel was covered with the stratified epithelium like that of 42-day rats. The present study has demonstrated that interglobular dentine contains sWGA-binding glycoconjugates and the formation of the interglobular dentine is largely associated with the enamel maturation. These results suggest that matrix-to-cell interaction is important for the development of interglobular dentine.

Acetylglucosamine↗

Multinucleated giant cells recruited by implantation of octacalcium phosphate (OCP) in rat bone marrow share ultrastructural characteristics with osteoclasts.

The ultrastructural characteristics of multinucleated giant cells (MNGCs) on octacalcium phosphate (OCP) and hydroxyapatite (HA) were investigated in comparison with those of osteoclasts, when the synthetic OCP and HA were implanted in rat bone marrow. The morphological difference in the MNGCs were shown between OCP and HA implants in 2 weeks after implantation. The MNGC on the implanted OCP (i-OCP) developed the ruffled border-like structure and the clear zone-like structure. The i-OCP was frayed where it was in contact with the ruffled border-like structure. The MNGC on the implanted HA (i-HA) developed the clear zone-like structure, whereas no ruffled border was seen. The surface of i-HA associated with the MNGC was smooth and not frayed. Bone formed directly on the OCP or HA implants. The interface between the i-OCP and bone matrix interdigitated, whereas that between the i-HA and bone matrix was comparatively smooth. The present study suggested that the i-OCP could be resorbed by the MNGCs which share some ultrastructural characteristics with osteoclasts.

Animals↗

Involvement of tumor necrosis factor-alpha, interleukin-1 beta, interleukin-8, and interleukin-1 receptor antagonist in acute lung injury caused by local Shwartzman reaction.

A local Shwartzman reaction (LSR) was prepared in rabbit lung as a model of acute lung injury. To induce LSR, intratracheal injection of lipopolysaccharide (LPS) 10 micrograms into the lower lobe of the right lung, followed 24 h later by i.v. injection of LPS (10 micrograms/kg). In the lung with the LSR, myeloperoxidase activity, representing neutrophil accumulation, peaked at 1-2 h and was sustained for 48 h after challenge with i.v. LPS. The lung water content peaked at 12 h, and decreased gradually. Histological findings showed diffuse interstitial widening, intra-alveolar leukocyte infiltration with hemorrhage, and alveolar exudate formation. The production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), interleukin-8 (IL-8), and IL-1 receptor antagonist (IL-1 Ra) in the lung was analyzed. TNF-alpha first elevated and peaked at 0.5 h (66.5 +/- 16.7 ng/g.lung), subsequently, IL-1 beta and IL-8 increased and peaked at 2 h (17.8 +/- 3.4 ng/g.lung and 336.9 +/- 49.6 ng/g.lung, respectively). IL-1Ra was present even before the challenge, and the production increased to show a dual peak (0.5 h, 1.5 +/- 0.2 micrograms/g.lung; and 2 h, 1.6 +/- 0.1 micrograms/g.lung), and a large concentration of IL-1Ra was sustained for 48 h. Immunohistochemistry showed that the cellular source of these cytokines was alveolar macrophages and infiltrating neutrophils. Thus, disclosing the kinetics of the generation of cytokines led to a better understanding of their roles, namely TNF-alpha as an initiator, IL-1 and IL-8 as amplifier and effector, and IL-1Ra as regulator of the intensity of acute inflammation.

Animals↗

Expression of osteocalcin in cementoblasts forming acellular cementum.

To determine the phenotypic expression of cementoblasts responsible for acellular cementum, an immunohistochemical study was performed using a polyclonal antibody raised against the aminoterminal peptide of rat osteocalcin (OC). Maxillary first molars of Wistar male rats aged 2 and 3 wk were used for observations. Serial sections of decalcified paraffin embedded specimens were stained either with hematoxylin and eosin or with the anti-OC antibody. In 2-wk-old rats, apical roots were lined with the epithelial root sheath. A thin layer of acellular cementum was seen at most of the root surface, but was not seen near to root apex. In 3-wk-old rats, cellular cementum began to be formed at root apex, and acellular cementum became more thick than in 2-wk-old rats. Acellular and cellular cementum were lined with the fibroblast-like cells. Osteocalcin staining was detected in cells lining root surface in both 2- and 3-wk-old rats. Almost all cells lining cellular cementum were positive for OC. In contrast OC positive cells lining acellular cementum and root surface devoid of cementum appeared at a specific site of the root. The cells at the interradicular area of root surface were positive but the cells at the outer area (the opposite side of the interradicular area) were negative for OC. Osteoblasts and odontoblasts were positive with the antibody. The present results suggest that the OC expression of cementoblasts forming acellular cementum is similar to that of cells forming cellular cementum as well as osteoblasts and odontoblasts, and has a role for calcification of acellular cementum.

Animals↗

Age-related changes in the localization of glycosaminoglycans in condylar cartilage of the mandible in rats.

There is little information available regarding the morphological and biomolecular characteristics of mandibular condylar cartilage. The purpose of this study was to determine the age-related changes in the morphology and immunolocalization of glycosaminoglycans (GAGs) in mandibular condyles. The mandibular condylar cartilages from 4-, 8-, 16-, 32-, and 64-week-old Wistar male rats were examined to verify the localization of chondroitin-4-sulfate (Ch-4S), chondroitin-6-sulfate (Ch-6S) and keratan sulfate (KS) using an indirect immunofluorescent technique with three monoclonal antibodies for glycosaminoglycans, 2-B-6, 3-B-3 and 5-D-4, respectively. Morphologically, the condylar cartilage was a growth cartilage during growing periods, began to differentiate into articular cartilage from the central area of 16-week-old condyles, and became mature articular cartilage at 32 weeks of age. A regional difference was found in the morphological features and distribution of GAGs between the anterior, central, postero-superior and posterior areas of the condyles at each age. The immunohistochemical localizations of these three glycosaminoglycans showed age-related, morphology-dependent changes, from growth cartilage to articular cartilage-like cartilage. Immunoreactions for all of the antibodies decreased progressively with age in the interterritorial matrix, while the pericellular and territorial matrix in the condylar cartilage of the mandible maintained relatively higher immunoreactivity. In conclusion, age-related and regional differences in the localization of glycosaminoglycans Ch-4S, Ch-6S, and KS were found in the mandibular condyles in rats, and these changes are believed to be related to functional and developmental requirements.

Aging↗

Chondrocytes synthesize type I collagen and accumulate the protein in the matrix during development of rat tibial articular cartilage.

The present study was designed to investigate whether or not chondrocytes in articular cartilage express type I collagen in vivo under physiological conditions. Expressions of the gene and the phenotype of type I collagen were examined in rat tibial articular cartilage in the knee joint during development. Knee joints of Wistar rats at 1, 5, and 11 weeks postnatal were fixed in 4% paraformaldehyde with or without 0.5% glutaraldehyde and decalcified in 10% EDTA. After the specimens were embedded in paraffin and serial sections made, adjacent sections were processed for immunohistochemistry and in situ hybridization for type I collagen. The epiphysis of the tibia was composed of cartilage in week- 1 rats. Formation of articular cartilage was in progress in week 5 as endochondral ossification proceeded and was completed in week 11. Anti-type I collagen antibody stained only the superficial area of the epiphysis in week 1, but the immunoreactivity was expanded into the deeper region of the articular cartilage with development in weeks 5 and 11. Hybridization signals for pro-alpha 1 (I) collagen were seen in some of chondrocytes in the epiphysis of the week-1 tibia. The most intense signals were identified in chondrocytes in week 5 and the signals appeared weaker in week 11. The present study demonstrated that chondrocytes synthesize type I collagen and accumulate the protein in the matrix during development of the articular cartilage.

Age Factors↗

Effects of expansive force on the differentiation of midpalatal suture cartilage in rats.

In an attempt to clarify the effects of biomechanical tensional force on chondrogenic and osteogenic differentiation of secondary cartilage, the midpalatal sutures of 4-week-old Wistar male rats were expanded by orthodontic wires which applied 20 g force for 4, 7, 10, and 14 days. The differentiation pathways in the midpalatal suture cartilage were examined by immunohistochemistry for osteocalcin, type I and type II collagen, and von Kossa histochemistry. Although the midpalatal sutures of the control animals consisted mainly of two separate secondary cartilages with mesenchyme-like cells at their midlines, type I collagen-rich fibrous tissue began to appear at day 4 and increased at the midline of the cartilage with days of experiment. At the end of the experiment, type I collagen-rich and calcified bone matrix appeared at the boundary between the precartilaginous and the cartilaginous cell layers. Most of the cartilaginous tissues were separated from each other and the midpalatal suture was replaced by osteocalcin-positive intramembranous bone and fibrous sutural tissue. These results strongly suggest that tensional force changed the phenotypic expression of collagenous components in secondary cartilage, which may reflect the differentiation pathway of osteochondro progenitor cells.

Animals↗

Initiation of alveolar ridge augmentation in the rat mandible by subperiosteal implantation of octacalcium phosphate.

The study was designed to investigate the process of bone formation caused by implantation of octacalcium phosphate as well as stability of the bone formed at the alveolar ridge. Synthetic octacalcium phosphate was implanted into a subperiosteal pocket in the rat mandible. Bone formation at the alveolar ridge was examined radiographically and histologically between 1 and 48 week(s) after implantation. Radiopacity of the octacalcium phosphate implant became obvious in week 2. Osteogenesis was initiated from the bone surface near the implantation site and multinucleated giant cells appeared on the implanted octacalcium phosphate in week 1. More apposition of new bone was observed on the implanted octacalcium phosphate in week 2 or later. Some implants were directly enclosed by newly formed bone and no cellular component was seen between the implant and the bone matrix. Many octacalcium phosphate implants were enclosed by bone, whereas the augmented ridge was not seen radiographically in week 24 or later. If its persistence can be improved, octacalcium phosphate could be used to augment atrophic alveolar ridges.

Alveolar Process↗

Localization of glycosaminoglycans in periodontal ligament during physiological and experimental tooth movement.

Localization of chondroitin sulphates in periodontal ligaments (PDL) of rat molar roots during physiological and experimental tooth movement were analysed immunohistochemically with the use of monoclonal antibodies, 3B3 and 2B6, specific to chondroitin 6-sulphate (CH-6S) and chondroitin 4-sulphate/dermatan sulfate (CH-4S/DS), respectively. The maxillary first molars of experimental animals were forced to move laterally with a 10 g weight by U-shaped wires for 3 and 7 d. In control animals, 3B3 epitope was seen in the PDL near to the bone surface facing the distal half of roots, which corresponded to the compressive side during physiological tooth movement. Immunoreactivity for 2B6 was weak or negative in the PDL. Both epitopes were present at osteoid, precementum, lacunae and canaliculli of osteocytes and cementocytes. In 3-d-treated animals, the early stage of hyalinization characterized with visible cells and fibres was observed in the PDL at the buccal side of the mesial root, which showed intense immunoreactivity for 3B3. Further 3B3 positive area seen in control animals changed its position from the distal to the buccal side of the PDL. Immunoreactivity for 2B6 did not change in the PDL of 3-d-treated animals. In 7-d-treated animals, the typical hyalinization characterized with no visible cells and fibres was seen in the PDL at the buccal sides of both mesial and disto-buccal roots, where both epitopes were present at the peripheral part of the tissue. Observation of serial sections suggested that the 3B3-positive area was present at the peripheral part of the 2B6-positive area. The present results suggest that the expression of CH-6S is related to the compressive force in non-hyalinized and hyalinized PDL, whereas that of CH-4S/DS is not influenced by the mechanical stress.

Alveolar Process↗