Structural variation of type XII collagen at its carboxyl-terminal NC1 domain generated by tissue-specific alternative splicing.
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This paper reports the identification of two structural variations in the NC1 domain of rat and mouse type XII collagen. The long NC1 domain encoding 74 amino acids showed homology to chicken type XII and XIV collagens. The short NC1 domain was composed of 19 amino acids. Through genomic DNA analyses, two alternative exons were identified, each of which contained the variable NC1 sequence. With the amino-terminal NC3 splicing alternatives, we propose here a new descriptive nomenclature: types XIIA-1 and XIIB-1 which include a long NC1 sequence encoded by exon 1 (from the 3'-end), and types XIIA-2 and XIIB-2 which include a short NC1 sequence encoded by exon 2. Types XIIA-1 and XIIB-1, the predominant transcripts in 15-day old mouse embryos, showed decreased expression in 17-day old embryos when type XIIB-2 expression was sustained at constant levels. In adult mice, type XIIB-1 associates with ligament and tendon, whereas type XIIB-2 is expressed in various other tissues. The long NC1 domain contains an extended acidic region (pI = 3.4) followed by a terminal basic region (pI = 13.8). Because the short NC1 domain lacks these features, structural variations in the type XII collagen NC1 domain suggests different functional roles in a tissue-specific fashion. (+info)
Bone wound healing after maxillary molar extraction in ovariectomized aged rats: quantitative backscattered electron image analysis.
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The processes of bone wound healing after maxillary molar extraction in ovariectomized aged rats were examined by means of quantitative backscattered electron image analysis and energy-dispersive X-ray microanalysis. Six-month-old female rats were either sham-operated or underwent bilateral ovariectomy (OVX), and 60 days postoperatively, the maxillary first molars were extracted. On post-extraction days 7, 30, and 60, the dissected and resin-embedded maxillae were micromilled in the transverse direction through the extracted alveolar sockets, and new bone formation on the buccal maxillary bone surface and within the extracted alveolar sockets was examined. In both sham-operated control and OVX rats, new bone formation was recognized on the buccal bone surface, as well as within the extracted sockets, and increased daily through to day 60. In comparison to sham-operated controls, new bone formation in OVX rats was significantly decreased both on the buccal bone surface and within the extracted sockets. Our results suggest that bone wound healing by new bone formation after maxillary molar extraction is significantly decreased in OVX-induced osteoporosis. (+info)
Transplantation of labeled periodontal ligament cells promotes regeneration of alveolar bone.
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Regeneration of damaged periodontal tissues is mediated by periodontal cells, but a major sub-population comprises highly differentiated cells that do not renew. To overcome the loss of specialized cell types caused by disease, various therapeutic approaches including cell transplants have been developed to promote cell re-population in periodontal tissues. As previous transplantation studies used unlabeled cells, that are indistinguishable from host cells, it has been difficult to assess the contributions of transplanted cells to the healing processes. To track the fate and differentiation of rat periodontal cells transplanted into periodontal wounds, we used collagen-coated fluorescent beads as a permanent endocytosed marker, or cells constitutively expressing beta-galactosidase. We assessed osteogenic cell differentiation with immunohistochemical staining for osteopontin and bone sialoprotein. Cells were transplanted into periodontal wounds created in Sprague--Dawley male rats that are null for beta-galactosidase. Defects were allowed to heal spontaneously (controls), or were closed with collagen implants mixed with beta-galactosidase-positive (Lac-Z) periodontal cells, or closed with collagen implants mixed with periodontal cells loaded with fluorescent beads. Animals were killed at 1 and 2 weeks after surgery and tissues were prepared for morphometric assessment and immunostaining for osteopontin (OPN) and bone sialoprotein (BSP). Transplanted cells were easily distinguished by fluorescent beads or by beta-galactosidase-positive expression and were distributed throughout the regenerating periodontal ligament (PL) and alveolar bone. At 1 week after wounding, animals treated with beta-galactosidase-positive cells exhibited a slightly higher percentage of labeled cells in the PL compared with the fluorescent bead-labeled cell implant group (2% vs. 1% respectively; P > 0.2). At Week 2 percentages of labeled cells were slightly increased in the regenerating PL (approximately 3% for both groups, P > 0.2). In regenerating alveolar bone at 1 week, animals that were treated with beta-galactosidase-positive cells and fluorescent bead-loaded cells exhibited approximately 30% and 25% of labeled cells respectively. At 2 weeks after wounding there was an increase in the percentage of transplanted beta-galactosidase-positive cells (approximately 39% at week 2; P < 0.05), but not of transplanted cells with fluorescent beads (approximately 25% at week 2). In sites with transplanted cells there were higher percentages of OPN positive and BSP positive cells in nascent bone and more newly formed bone than in controls (>40%; P < 0.05). Transplantation of beta-galactosidase-positive cells or cells loaded with fluorescent beads is a useful method for assessing the fate and differentiation of periodontal cells in vivo. Fluorescent beads, however, are diluted at mitosis and this method underestimates the percentage of transplanted cells. As transplanted periodontal cells in both groups promoted regeneration of alveolar bone, cell transplantation could improve the restoration of periodontium destroyed by periodontitis. (+info)
Acute effects of ovariectomy on wound healing of alveolar bone after maxillary molar extraction in aged rats.
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Acute effects of ovariectomy on the bone wound healing processes after maxillary molar extraction in aged rats were examined by means of quantitative scanning electron microscopy (SEM), backscattered electron image (BSE) analysis and energy-dispersive X-ray (EDX) microanalysis. Six-month-old female rats underwent either sham operation or bilateral ovariectomy, and 7 days postoperatively, the maxillary first molars were extracted. On post-extraction days 7, 30 and 60, the dissected maxillary bone surfaces were examined by SEM to reveal the bone formative and resorptive areas around the extracted alveolar sockets. In addition, the resin-embedded maxillae were micromilled in the transverse direction through the extracted alveolar sockets, and the newly-formed bone mass on the buccal bone surfaces and within the extracted sockets was examined by BSE analysis. Compared with sham-operated controls, the extent of newly-formed bone mass on the buccal bone surfaces in OVX rats was significantly decreased, due to increased bone resorption. On the other hand, new bone formation within the extracted sockets was similar in the experimental groups. In EDX microanalysis of these newly-formed bone matrices, both Ca and P weight % and Ca/P molar ratio were similar in the experimental groups. Our results suggest that 1) acute estrogen deficiency induced by ovariectomy stimulates sustained bone resorption, but has less effect on bone formation, and 2) bone wound healing after maxillary molar extraction within extracted alveolar sockets is not significantly delayed by ovariectomy, but bony support by newly-formed bone mass on the maxillary bone surfaces at the buccal side of the extracted sockets is significantly decreased, due to increased bone resorption. (+info)
Histologic study of use of microfibrillar collagen hemostat in rat dental sockets.
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The aim of this paper was to evaluate if the placement of microfibrillar collagen hemostat (MCH) into a dental socket interfered with healing. General anesthesia was administered to 30 adult male Albinus Wistar rats and the maxillary right central incisor was extracted. In the control group after each tooth was extracted, the socket was sutured. In the MCH group after each tooth was extracted, MCH was placed into the socket before suturing. Postoperatively, 5 animals were sacrificed from each group at 7, 21 and 28 days. The right maxilla was removed from each animal and histologic slides were stained with Masson's trichromic and hematoxylin and eosin. Quantitative and qualitative analyses were done. The percentage of bone area in the dental socket was quantified using the Image Lab 98 image analysis system. The bone area formation for the control and MCH groups was: 8.1% and 3.3% at 7 days, 34.4% and 33% at 21 days and 41% and 41.3% at 28 days, respectively. We concluded that MCH interferes with the beginning of dental socket healing but does not interfere with the final healing of the dental socket. (+info)
Restoration of mechanical strength and morphological features of the periodontal ligament following orthodontic retention in the rat mandibular first molar.
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Biomechanical properties and morphological features of the periodontal ligament (PDL) in the rat mandibular molars were examined during orthodontic retention. Seventy-three male rats of the Wistar strain, 8 weeks of age, were used for biomechanical analysis and six rats for morphological analysis. An elastic band was inserted between the mandibular first and second molars for 4 days; after removal of the elastic band the interdental space was filled with resin for 4 and 8 days. The maximum shear stress, tangent modulus, and failure strain energy density of the PDL of the first molar in the experimental animals decreased markedly following application of an orthodontic force. They increased rapidly and were restored completely to the control levels by the 8th day after retention. Light microscopy showed severe compression and extension of the PDL in the experimental animals on the 8th day after retention. Birefringent collagen fibre bundles running across the compressed and expanded PDL were observed, although they appeared to be thinner with less insertions into the alveolar bone or cementum in the experimental animals than in the controls. This suggests that the periodontal collagen fibres were partially reorganized and rearranged during retention. The reorganization and rearrangement of periodontal collagen fibres seemed to be partly related to the restoration of mechanical strength of the rat molar PDL during the 8 days of retention. (+info)
Biological characteristics of cell lines of human dental alveolus.
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OBJECTIVE: To investigate the biological characteristics of cell lines of healthy and diseased human dental alveoli. METHODS: Primary cell lines from either healthy or diseased human dental alveoli were obtained. Two cell lines, H-258 and H-171 derived from healthy and diseased human tissues respectively, were selected for morphological study and research on their growth and aging, using cell counting, and histochemical and immunohistochemical staining. RESULTS: Primary cell lines were successfully established from innormal dental alveoli. After freezing and thawing for three times, cell growth was continued and no morphological alterations were observed. The doubling time was 53.4 hours and mean division index (MDI) was 4 per thousand. Cells were kept normal after twenty generations with no obvious reduction of doubling time and MDI. Of twenty-six primary cell lines derived from healthy human dental alveoli, only three cell lines achieved generation. After freezing and thawing for twice, cultured cells were still alive at a decreased growth speed, with doubling time of 85.9 hours and MDI of 3 per thousand. Both cell lines, H-171 and H-258, shared the characteristics of osteoblast. CONCLUSIONS: Primary cell lines of diseased human dental alveoli show greater growth potential. All cell lines of dental alveoli share characteristics of osteoblast. The technique we developed may be put into practice for the treatment of abnormal dental alveoli. (+info)
Two- and three-dimensional sonographic assessment of the fetal face. 2. Analysis of cleft lip, alveolus and palate.
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OBJECTIVES: To describe the sonographic appearance of cleft lip with or without cleft palate (CL +/- P) using two-dimensional and three-dimensional (3D) ultrasound imaging. Also, to evaluate the accuracy of ultrasound to delineate with precision the bony extent of facial clefts, i.e. to differentiate clefts limited to the lips, or extending to the alveolus/premaxilla or the secondary palate. METHODS: This was a retrospective study based on the examination of fetuses diagnosed with an isolated CL +/- P. Cases included were either discovered at systematic screening or referred for further investigation. Clefts were characterized by their precise anatomical location and extent. The defect could include a cleft lip (CL), a cleft alveolus (CA), or a cleft of the secondary palate (CSP). RESULTS: We analyzed 96 cases of CL +/- P. The mean gestational age at examination was 28.2 +/- 4.1 weeks. The sonographic appearance of CL, CA, and CSP was depicted. Strict concordance of the sonographic report with the anatomical defect was present in 84 cases (87.5%). In eight cases, the severity of the cleft was underestimated: three cases of CA, four of CA + CSP and one of CSP were missed. In four cases, the cleft was overestimated as CA was incorrectly suspected. CONCLUSIONS: Systematic screening with sonography to detect prenatally CL +/- P requires the imaging of at least the mid-sagittal and the anterior coronal 'nose-mouth' views. Once the presence of a facial cleft is suspected, the three reference orthogonal planes are imaged in order to characterize the anatomical defect, and for each plane, the serial scans are thoroughly examined. This protocol allows precise delineation of the defect. Inclusion of 3D and 4D ultrasound imaging in the examination protocol allows easier and more rapid screening and more precise evaluation of the different cleft constituents. (+info)