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The nuclear lamina and its functions in the nucleus.

The nuclear lamina is a structure near the inner nuclear membrane and the peripheral chromatin. It is composed of lamins, which are also present in the nuclear interior, and lamin-associated proteins. The increasing number of proteins that interact with lamins and the compound interactions between these proteins and chromatin-associated proteins make the nuclear lamina a highly complex but also a very exciting structure. The nuclear lamina is an essential component of metazoan cells. It is involved in most nuclear activities including DNA replication, RNA transcription, nuclear and chromatin organization, cell cycle regulation, cell development and differentiation, nuclear migration, and apoptosis. Specific mutations in nuclear lamina genes cause a wide range of heritable human diseases. These diseases include Emery-Dreifuss muscular dystrophy, limb girdle muscular dystrophy, dilated cardiomyopathy (DCM) with conduction system disease, familial partial lipodystrophy (FPLD), autosomal recessive axonal neuropathy (Charcot-Marie-Tooth disorder type 2, CMT2), mandibuloacral dysplasia (MAD), Hutchison Gilford Progeria syndrome (HGS), Greenberg Skeletal Dysplasia, and Pelger-Huet anomaly (PHA). Genetic analyses in Caenorhabditis elegans, Drosophila, and mice show new insights into the functions of the nuclear lamina, and recent structural analyses have begun to unravel the molecular structure and assembly of lamins and their associated proteins.

Animals↗

Supra-alveolar periodontal healing of auto- and allotransplanted teeth in monkeys.

Although intra-alveolar healing of allotransplanted teeth often shows pathological changes due to rejection of the tooth graft, previous clinical findings seem to indicate that the supra-alveolar part of the allografts heals differently, which is of significant prognostic value. The aim of the present study was to determine the healing of the cervical part of the periodontium of auto- and allotransplanted mature teeth in monkeys. In 4 adult, immunologically unmatched, green vervet monkeys 16 mature permanent maxillary and mandibular incisors were extracted, endodontically treated and either auto- or allotransplanted; 8 untreated neighbouring incisors served as controls. Block biopsies including part of the tooth and the adjacent vestibular tissues were obtained after a healing period of 8 weeks. The biopsies were decalcified in EDTA and embedded in Epon. In semi-thin section (1.5 microns) the morphology of the supra-alveolar periodontal tissues was analysed in the light microscope. The orientation of the connective tissue fibres close to the root surface was found to be more parallel to the root surface of allotransplants compared with autografts, while in the control teeth the fibres were generally oriented perpendicular to the root surface. Resorption of the root was seldom found in the supra-alveolar portions of any of the transplanted teeth. The junctional epithelium had migrated slightly apically onto the root surfaces of allotransplanted teeth. The gingival connective tissue adjacent to allotransplanted teeth showed a significantly increased number of lymphocytes compared with autografts and controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Guided tissue regeneration. An experimental procedure in beagle dogs.

This preliminary study examined the healing following an experimental procedure designed to facilitate coronal migration of progenitor cells from the periodontal ligament circumferentially on roots of premolar teeth in beagle dogs. Mucoperiosteal flaps were reflected on the buccal and lingual aspects of premolars in six beagle dogs with periodontal disease. Following root preparation, pieces of orthodontic wire were placed interproximally on the crowns to bridge the spaces between teeth. Biobrane, a synthetic membrane bonded to a knitted nylon fabric and coated with collagen, was placed as a physical barrier between the roots and the flaps to be replaced. The membrane extended as a single piece from the cementoenamel junction (CEJ) to overlap the crest of alveolar bone by 3 to 4 mm on both the buccal and lingual surfaces of the three premolars in each quadrant. The membrane was attached to the crowns at the CEJ with resin. The flaps were replaced and sutured. Postoperative care included plaque control and the membranes were removed after 5 weeks. The dogs were sacrificed to provide observation periods of 8 and 16 weeks after placement of membranes. Histologic examination revealed new connective tissue attachment in the apical part of the 8- and 16-week experimental specimens. Some experimental specimens showed new attachment up to 2.94 mm while others showed a long junctional epithelium (JE). Root resorption was also seen in some specimens. These preliminary findings suggest that placement of physical barriers between root surface and flaps may be beneficial in facilitating coronal migration of progenitor cells from the periodontal ligament.

Animals↗

[The role of cell adhesion molecules in the formation of periapical granulomas].

Cell adhesion molecules (CAMs) are cell surface proteins involved in the binding of cells, usually leukocytes, to each other, to endothelial cells, or to extracellular matrix. Specific signals produced in response to wounding and infection control the expression and activation of certain of these adhesion molecules. The interactions and responses then initiated by binding of these CAMs to their receptors/ligands play important roles in the mediation of the inflammatory and immune reactions that consult one line of the body's defense against these insults. Most of the CAMs characterized so far fall into three general families of proteins: the immunoglobulin (Ig) superfamily, the integrin family, or the selectine family. Recent studies have indicated that selectins (E,L,P) are implicated in cell trafficking, an important aspect of inflammation-related process. Regulation of white blood cell trafficking from the blood vascular compartment to regions of pathogenic exposure is one of the most important functions of the immune system. The distinct phases of leukocyte migration include: rolling, activation, firm adhesion, transendothelial migration and subendothelial migration. The selectins have been implicated in the first step of this cascade. An inflammatory response is first evoked in the pulpal tissue in an attempt to neutralize the injurious agent and to dispose of damaged tissue and cells. The pulpal vessels dilate and blood flow to the tooth increases. At the same time, permeability of the vessels increases allowing leakage of fluid and leukocytes into the tissue.

Cell Adhesion Molecules↗

Actin-associated proteins in ameloblast differentiation.

Focal contacts are systems of adherens junctions of the cell-extracellular matrix type, which allow the transfer of fundamental signals from the extracellular matrix to nuclear compartments, capable of regulating adhesion, proliferation, migration and differentiation of cells. Recently, many authors have concentrated their attention on epitheliomesenchymal interactions which guide organogenesis of dental germ, identifying numerous growth and differentiation factors and having the inner enamel epithelial cells of the enamel organ as a target. Given that the two cellular compartments in their tooth germ are separated by a basal membrane and by an extracellular matrix, which touches it, we wanted to evaluate the presence of focal contacts through the identification of talin and vinculin, proteins of the actin-associated protein complex. In this study we utilized the hemimandibles of young Wistar rats and we extracted the related odontogenic tooth organs present at their apical end. Specimens are processed with antibody against vinculin and talin. Results show that these junctional system proteins are present at the apical poles of both cellular compartments suggesting that putative epithelial-mesenchymal interactions, other than marker molecules, may use focal contacts as a system for transmission of signals.

Actins↗

Taxonomy and phylogenetic relationships of early Miocene catarrhines from Sihong, China.

Paleontological investigations at sites in Sihong County, Jiangsu Province, China since 1981 have yielded a sizeable collection of previously undescribed fossil catarrhines from the Xiacaowan Formation. The associated vertebrate fauna indicates a late early Miocene age (correlating with MN 4, late Orleanian of Europe, approximately 17-18 Ma), which establishes the Sihong primates as the earliest known catarrhines from Eurasia. The fossil primates are assigned to two species: Dionysopithecus shuangouensis Li, 1978 and Platodontopithecus jianghuaiensis Gu & Lin, 1983. Although the new material from Sihong consists mainly of isolated teeth, it does provide important new information on the anatomy of Dionysopithecus and Platodontopithecus that helps to clarify their phylogenetic and taxonomic status. Previous studies have suggested that the Sihong catarrhines might be closely related to the proconsulids from the early Miocene of East Africa. However, with more extensive material available for comparison, the Sihong primates can now be shown to share a number of key derived features with pliopithecids. This new evidence helps to resolve a longstanding problem concerning the origins of the Pliopithecidae. It was previously considered that specialized pliopithecids migrated into Europe during MN 5, originating from an unknown antecedent and location in Africa. Recognition that the Sihong primates have affinities with pliopithecids, but are more primitive, suggests that the initial differentiation and diversification of the clade may have taken place in Asia rather than Africa. The earliest Eurasian catarrhines probably migrated into tropical Asia as part of a major faunal interchange with Africa that occurred during MN 3.

Animals↗

The effects of partial demineralization and fibronectin on migration and growth of gingival epithelial cells on cementum in vitro.

The capacity of mineralized cementum to support epithelial cell migration and growth and the effect that fibronectin and partial demineralization of cementum have on these processes were assessed in vitro. Dog gingival explants, 1 X 2 mm, were cultured on the cementum surfaces of pig root pieces in a defined medium consisting of DMEM and F12 (1V/1V), transferrin, insulin, epidermal growth factor, cortisone, selenium, and high-density lipoprotein. Sixty root pieces were divided into four equal groups according to the treatment: (1) untreated mineralized cementum; (2) treated with 5 micrograms of fibronectin; (3) partially demineralized in 18% EDTA for 30 min; and (4) both partially demineralized and fibronectin-treated as above. Epithelial cell migration and growth on each of the four differently treated cementum surfaces were assessed histomorphometrically by means of scanning electron microscopy. The defined culture medium supported the selective migration and growth of epithelial cells from the gingival explants onto the mineralized cementum. This was confirmed by the positive immunostaining of these cells with antikeratin antibodies. Partial demineralization of cementum inhibited epithelial migration and growth by 83% and 91%, respectively. Fibronectin treatment did not affect epithelial cell migration and growth on mineralized cementum, but it decreased the degree of epithelial cell migration and growth inhibition on partially demineralized cementum to 57% and 43%, respectively. The results indicate that: (i) mineralized cementum may consist of components that are recognized by gingival epithelial cells and support their migration and growth in vitro; (ii) these components can be removed by demineralization; and (iii) fibronectin partially restores epithelial cell migration and growth on partial demineralized cementum in vitro.

Animals↗

Role of osteopontin in bone remodeling caused by mechanical stress.

Changes in the number and proportion of osteopontin mRNA (Opn) expressing osteocytes and osteoclasts caused by the mechanical stress applied during experimental tooth movement were examined in the present study. Opn expression was detected in the osteocytes on the pressure side at the early stage, and gradually spread to those on the tension side and also to the osteoblasts and bone-lining cells in the alveolar bone. Only 3.3% of the osteocytes located on the pressure side expressed Opn in the interradicular septum of control rats; in contrast, the value was increased to 87.5% at 48 h after the initiation of tooth movement. These results indicate that these cells responded to mechanical stress loaded on the bone with expression of the osteopontin gene. Following the increased expression of Opn in these cells, a 17-fold greater number of osteoclasts compared with the control and numerous resorption pits were observed on the pressure side of the alveolar bone. Injection of arginine-glycine-aspartic acid-serine peptide but not that of arginine-glycine-glutamic acid-serine peptide strongly inhibited the increase in the number of osteoclasts. Furthermore, an in vitro migration assay demonstrated the chemotactic activity of osteopontin (OPN) on the precursor of osteoclasts. Our study strongly suggests that OPN is an important factor triggering bone remodeling caused by mechanical stress.

Animals↗

Cell and fiber attachment to demineralized dentin from normal root surfaces.

The study assessed connective tissue and epithelial responses to dentin specimens (obtained from normal roots of human teeth) after surface demineralization. Rectangular dental specimens with opposite faces of root and pulpal dentin were prepared from beneath root surfaces covered by periodontal ligament. One-half of the specimens were treated with citric acid, pH 1, for 3 minutes, while the remainder served as untreated control specimens. Specimens were implanted vertically into incisional wounds on the dorsal surface of rats with one end of the implant protruding through the skin. Four specimens in each group were available 1, 3, 5 and 10 days after implantation. Histologic and histometric analyses included counts of adhering cells, evaluation of connective tissue fiber relationships and assessment of epithelial migration. Analyses within each group comparing root and pulpal surfaces showed no differences between any of the parameters. Comparisons between experimental and control groups showed that demineralized surfaces had a greater number of cells attached, fiber attachment occurred and epithelial downgrowth was inhibited. The fiber attachment to experimental specimens differed morphologically from fiber attachment to normal root surfaces: the number of fibers attached per unit length and the diameter of attached fibers were significantly less on experimental specimens. Demineralized specimens at 10 days had a distinct eosinophilic surface zone. Surface demineralization of dentin predisposed toward a cell and fiber attachment system which inhibited migration of epithelium.

Animals↗

Spontaneous correction of pathologic migration following periodontal therapy.

Two case reports illustrate spontaneous correction of pathologic migration following periodontal therapy. In one case, the displaced teeth were repositioned without orthodontic appliances after nonsurgical and surgical periodontal treatment. In the other case, closure of a diastema occurred following only nonsurgical therapy. The predictability of these findings needs further investigation. A discussion of possible reasons for this type of tooth movement is included.

Diastema↗

Implant failure with spontaneous rapid exfoliation: case reports.

BACKGROUND: The predictability and success of endosseous dental implants has secured their place as a standard treatment modality. Nevertheless, a small number of implants will fail regardless of operator experience or clinically recognizable cause. This article presents two cases of early failure of two-stage dental implants manifesting in a spontaneous and rapid exfoliation. METHODS: Two healthy adult male patients received single-tooth, machined titanium implant fixtures during uncomplicated stage-one surgical procedures. RESULTS: After an uneventful early postoperative healing phase, both patients returned during the 4 to 6 week time period with failing implants. Clinical examination in both cases revealed the exfoliation of a fixture from the edentulous area with varying signs of soft tissue reaction in each instance. Radiographs revealed osteotomy sites along with the coronally migrating fixtures, with one of the exfoliated implants accompanied by apparent perifixtural radiolucency. In both cases, the implant fixtures were removed digitally without anesthesia. Subsequent medical follow-up was noncontributory, and ultimate healing was uneventful in both cases. Two separate attempts in the mandibular right second premolar region of one patient ended in similar failures despite the successful osseointegration of an identical fixture on the contralateral side. CONCLUSIONS: Few cases of early exfoliation of dental implants have been reported in the literature. Although the exact etiology of the early fixture loss in these two patients is unclear, the description of these cases can add to the published knowledge of dental implant failures and potentially lead to the uncovering and prevention of their causative mechanisms.

Adult↗

Effect of citric acid and lyophilized autologous plasma on healing following periodontal flap surgery in monkeys.

The purpose of this histologic, histometric, and autoradiographic study was to examine the effect of citric acid conditioning and lyophilized autologous plasma (LAP) application on healing following periodontal flap surgery. Mucoperiosteal flaps were elevated in six rhesus monkeys using the modified Widman flap procedure. A total of 24 quadrants were treated, each included the first and second premolar and first and second molar teeth. Cementum was removed from the exposed root surfaces, and reference notches were inscribed into the roots at the alveolar bone margin. Two treatment modalities were employed: (1) surgery plus citric acid conditioning, to serve as control and (2) surgery plus citric acid followed by LAP application (400 mg/ml saline). Flaps were returned to their preoperative positions and sutured. Animal sacrifices were scheduled to provide observations 3, 7, 14, 21, 28, and 45 days after surgery. Each monkey received an intravenous injection of tritiated thymidine, 1 microCi/gm of body weight, 1 hour before it was killed. Tissue specimens were processed for evaluation following standard procedures. Histologically, tissue sections were examined for: (1) proliferation and attachment of epithelium, (2) organization and maturation of periodontal fibers, (3) inflammatory cell types, (4) presence or absence of new cementum deposition, and (5) degree of vascularity of the tissues. For histometric evaluation, the radicular notches were used as reference points. The distances examined histometrically were: (1) from the root surface notch to the alveolar bone crest, (2) from the root surface notch to the apical extent of the junctional epithelium, and (3) from the free gingival margin to the apical extent of the junctional epithelium. For autoradiographic evaluation labeled cells were counted in five spatial compartments at 400 X magnification: (1) oral epithelium, (2) crevicular area, (3) supracrestal connective tissue, (4) coronal periodontal membrane, and (5) coronal bone marrow. For each the histometric and autoradiographic evaluation involved a total of 36 tissue sections per quadrant (9 sections per tooth). Tooth and quadrant means were obtained for each monkey. The plasma-treated and control quadrants were compared at each time point by the paired t test. N = 2 monkeys were used for each comparison. Histologic results showed that in teeth that were acid-conditioned after root planing, the epithelium often migrated apically reaching the radicular notch. Those teeth that were conditioned and subsequently treated with LAP demonstrated fiber attachment to the planed root surface and little or no epithelial downgrowth.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Coronal microleakage assessed by polymicrobial markers.

Studies have shown significant bacterial leakage following exposure of sealed root canals to artificial and natural saliva. The objective of this study was to determine contamination via bacterial migration in artificial saliva in endodontically treated teeth using different obturation techniques and sealers. A total of 144 extracted, human mandibular anterior teeth were cleaned and shaped to a #40 master apical file using hand and rotary instrumentation. One hundred and twenty teeth were divided into two experimental groups. The teeth in Group 1 were obturated with gutta-percha using lateral compaction and five different sealers, and the teeth in Group 2 were obturated with gutta-percha using thermomechanical compaction and five different sealers. The remaining 24 teeth were prepared as positive and negative controls. All specimens, except the negative controls, were inoculated every five days with Anaerobic streptococci (NCTC 9891) related to Peptostreptococcus micros and Prevotella intermedia (ATCC 25611). The contamination onset time was continuously recorded and turbid broths cultured for bacteria identification. The controls behaved as expected. Regardless of the combination between obturation techniques and different sealers, all broths became turbid during this experiment. The correlation between the obturation techniques and the sealers revealed statistical significance using ANOVA (p<0.0001), followed by the Duncan Multiple Range Test, which determined the ranking between these interactions. The combination of MicroSeal obturation technique with Ketac-Endo sealer allowed a slower rate of coronal-apical bacterial migration.

Analysis of Variance↗

The potential coronal migration of periodontal ligament tissue following experimental regeneration.

The aim of the present study was to determine if the periodontal ligament cells can migrate onto curretted root surfaces following an experimental regeneration procedure. Buccal mucoperiosteal flaps were elevated in all three premolar regions in 5 mongrel dogs. The buccal bone was reduced to approximately 7 mm from its original level on 30 roots. Before the elevated flaps were replaced and sutured, Gore-Tex filters were adjusted to cover exposed root surfaces. No membranes were placed over 10 root surfaces, which served as controls. After 7 weeks of healing, the animals were sacrificed and all the roots were examined for histological evaluation of contralateral experimental and control teeth. With polychrome staining, the new periodontal ligament fibers of the curetted root surfaces were dyed methyl blue and the adjacent gingival collagen fibers were dyed red. On the experimental surface, coronal migration of periodontal ligament cells and connective tissue attachment were identified over long distances of curetted root surface. Periodontal ligament spaces were more prevalent in experimental sites than in control root surfaces. The results of the experiment suggest that the periodontal ligament cells actually migrated onto the curetted root surfaces when a periodontal space was created by physical barriers.

Animals↗

The molecular elasticity of the extracellular matrix protein tenascin.

Extracellular matrix proteins are thought to provide a rigid mechanical anchor that supports and guides migrating and rolling cells. Here we examine the mechanical properties of the extracellular matrix protein tenascin by using atomic-force-microscopy techniques. Our results indicate that tenascin is an elastic protein. Single molecules of tenascin could be stretched to several times their resting length. Force-extension curves showed a saw-tooth pattern, with peaks of force at 137pN. These peaks were approximately 25 nm apart. Similar results have been obtained by study of titin. We also found similar results by studying recombinant tenascin fragments encompassing the 15 fibronectin type III domains of tenascin. This indicates that the extensibility of tenascin may be due to the stretch-induced unfolding of its fibronectin type III domains. Refolding of tenascin after stretching, observed when the force was reduced to near zero, showed a double-exponential recovery with time constants of 42 domains refolded per second and 0.5 domains per second. The former speed of refolding is more than twice as fast as any previously reported speed of refolding of a fibronectin type III domain. We suggest that the extensibility of the modular fibronectin type III region may be important in allowing tenascin-ligand bonds to persist over long extensions. These properties of fibronectin type III modules may be of widespread use in extracellular proteins containing such domain.

Alternative Splicing↗

Improved primary surgical and dental treatment of clefts.

The improved combination of surgical and dental teamwork in the primary treatment of clefts presented here is consistent with principles. In fact, this is a staged design for correction of classic clefts of the lip and palate that, based on biological principles, facilitates the continuance of the failed embryonic "migrations" toward a normal end point. Positioning of the alveolar segments, dissection of mucoperiosteum out of the cleft, and union of mucoperiosteum across the alveolar and anterior hard palate cleft make it possible to create a periosteal tunnel across the bony gap and set up a condition conducive to bone formation and eventual tooth eruption in the cleft area. Lip closure by adhesion reduces the tension of the primary lip closure and allows gentle molding until solidification of the arch occurs. Thus a complete cleft has been rendered an incomplete cleft. With a balanced, stabilized maxillary platform, the definitive lip and nose corrections can be carried to completion early (by 2 to 4 years of age). These planned actions bypass a persistent cleft, fistulas, raw areas, malposition of alveolar segments, and probably the necessity for later bone grafting. The only question not totally answered is the effect of this approach on final growth. Although most reports seem to indicate that growth has and will proceed within normal limits, another 10 years of careful follow-up is indicated and, in fact, is in progress.

Child↗

Behavior of odontoblasts and basal lamina of trypsin or EDTA-isolated mouse dental papillae in short-term culture.

Embryonic mouse first mandibular molars (day 18), containing the first overtly differentiated odontoblasts, were treated with EDTA or trypsin, to obtain isolated dental papillae. Trypsin dissociation was accompanied by disappearance of the basal lamina. EDTA-treatment removed the basal lamina from the basal surface of the epithelium, but left it associated with the dental papillae. EDTA- or trypsin-isolated dental papillae were grown for three to 24 h at the top of a plasma clot. Trypsin-isolated dental papillae rapidly lost their typical morphology, and the post-mitotic odontoblasts never became functional. EDTA-isolated dental papillae remained covered by the basal lamina for 15 to 18 h. During this period, the typical morphology was maintained, and post-mitotic odontoblasts secreted predentin. Preodontoblasts and odontoblasts went through the basal lamina and migrated at the outer surface of the basal lamina (i.e., at the side facing away from the enamel organ).

Animals↗

Changes in cell phenotype during regeneration of junctional epithelium of human gingiva in vitro.

The relationship between cell attachment and the phenotype of the attached oral epithelial cells was studied by comparing junctional epithelium (JE) with a culture model for JE in which epithelial cells form an equivalent organization of tissues. Gingival explants were cultured on either a high or a low protein-binding membrane. The cut edge of epithelium and connective tissue was placed on the membrane; epithelial cells migrated to form a sheet of tissue between the explant and the membrane substratum. Cells which grew in contact with the high protein-binding membrane attached to the substratum and assumed a cuboidal shape. With time in culture these cells showed a decrease in reactivity with antibodies to psi-3 antigen (an antigen associated with epithelial migration) and an increase in reactivity with antibodies to cytokeratin 19 (a marker for JE). Cells grown on the low protein-binding membrane did not exhibit changes in shape or antigens. Because similar features were found in the JE in vivo, it was concluded that the junctional cells which are directly attached to the tooth (DAT cells) have a nonmigratory phenotype that develops as a response to the tooth surface. Because the cells are in contact with a metabolically inert material the changes appear to be largely self-induced. The culture method allows studies on putative inducer molecules and on mechanisms which may control the phenotype of epithelial cells at the dentogingival interface.

Adult↗