Bundles Of Collagen Fibrils Research Articles

Morphology and biomechanics of the microfibrillar network of sea cucumber dermis

The principal component of the body wall of the sea cucumber Cucumaria frondosa is a dermis consisting of collagen fibrils, microfibrils, proteoglycans and other soluble and insoluble components. A major structural constituent of the dermis is a network of 10­14 nm diameter microfibrils, which surrounds and penetrates bundles of collagen fibrils. This network has been extracted and purified using guanidine and bacterial collagenase. Tensile testing of the microfibrillar network in artificial sea water demonstrates that it is reversibly extensible up to approximately 300 % of its initial length. It behaves like a viscoelastic solid, having a long-range elastic component as well as a time-dependent viscous component. Reduction and alkylation of the cysteine residues in the network do not change its breaking strain or strength, but greatly increase the compliance of the network until, near the breaking strain, the tensile resistance rapidly increases. These data suggest that the strength of the network is due to non-reducible crosslinks, while its elasticity is dependent upon disulfide bonds. In deionized water, the network becomes swollen and, although it remains elastic, is much more compliant than when tested in artificial sea water. Examination of whole tissues and purified networks with the electron microscope reveals structures similar to vertebrate fibrillin-containing microfibrils. Considering that the dermis of C. frondosa is a mechanically mutable tissue in which elongation is accompanied by the sliding of collagen fibrils past one another, the microfibrillar network may act to maintain the orientation of fibrillar components during movement and may also provide a long-range restoring force.

Electron microscopic study on interodontoblastic collagen fibrils in amputated canine dental pulp

The purpose of this research was to study the presence and the ultrastructural features of interodontoblastic collagen fibrils and their contribution to the formation of reparative dentin in dog incisors and premolars for a period of 30 days following experimental pulpotomy. On the seventy day after pulp exposure and capping with calcium hydroxide, short cylindrical-shaped cells collected at the coronal end of the vital pulp tissue. Many collagen fibrils were synthesized in the intercellular spaces and in the direction of the long axis of these cells. On the fourteenth day, large bundles of collagen fibrils were observed in the intercellular spaces of young odontoblasts. These bundles spread out in a fan-shaped arrangement. On the thirtieth day, the odontoblasts formed a tubular dentin matrix. Small numbers of twisted collagen bundles crossed the distal junctional complex of the odontoblast cell bodies and entered the dentin matrix.

Three dimensional architecture of collagen fibrils in the corpus cavernosum of the crab-eating monkey.

The three-dimensional architecture of the collagen fibrils in the corpus cavernosum of the crab-eating monkey was studied by scanning electron microscope, after digestion of cellular and some extracellular elements by NaOH. The tunica was made up of undulating bundles of collagen fibrils which were arranged in inner circular, middle intermingled and outer longitudinal layers. The trabecula appeared as numerous cylindrical tubes which had once accommodated individual smooth muscle fibers. The outer surface of the trabecula was covered with a sheath woven from twisting bundles of collagen fibrils. These architectural features of the collagen fibrils seem to be related to fluctuations in the diameter and shape of the corpus during erection and detumescence.

Radial organization of interstitial exchange pathway and influence of collagen in synovium

The synovial intercellular space is the path by which water, nutrients, cytokines, and macromolecules enter and leave the joint cavity. In this study two structural factors influencing synovial permeability were quantified by morphometry (Delesse's principle) of synovial electronmicrographs (rabbit knee), namely interstitial volume fraction Vv.1 and the fraction of the interstitium obstructed by collagen fibrils. Mean Vv.1 across the full thickness was 0.66 +/- 0.03 SEM (n = 11); but Vv.1 actually varied systematically with depth normal to the surface, increasing nonlinearly from 0.40 +/- 0.04 (n = 5 joints) near the free surface to 0.92 +/- 0.02 near the subsynovial interface. Tending to offset this increase in transport space, however, the space "blocked" by collagen fibrils also increased nonlinearly with depth. Bundles of collagen fibrils occupied 13.6 +/- 2.4% of interstitial volume close to the free surface but 49 +/- 4.8% near the subsynovial surface (full-thickness average, 40.5 +/- 3.5%), with fibrils accounting for 48.6-57.1% of the bundle space. Because of the two counteracting compositional gradients, the space available for fibril-excluded transport (hydraulic flow and macromolecular diffusion) was relatively constant > 4 microns below the surface but constricted at the synovium-cavity interface. The space available to extracellular polymers was only 51-53% of tissue volume, raising their effective concentration and hence the lining's resistance to flow and ability to confine the synovial fluid.

Evolution of three patterns of intra-alveolar fibrosis produced by bleomycin in rats.

In pulmonary fibrosis, it is known that fibrotic changes develop in the intra-alveolar spaces and that intra-alveolar fibrosis can be classified into three patterns, namely intra-alveolar buds, mural incorporation and obliterative changes. In order to clarify the evolution of intra-alveolar fibrosis, immunohistochemical studies of extracellular matrix proteins and electron microscopic observations were made of the lungs of rats given a single intratracheal instillation of bleomycin. All three patterns of fibrosis developed in this model. Intra-alveolar buds changed into globular lesions with dense collagen deposition, the surface of which was covered by alveolar epithelium. Electron microscopy revealed that the buds often contained spiraling collagen fibrils and numerous microfibrils, but not mature elastic fibres, beneath the regenerating epithelial lining cells; the epithelial basement membranes were discontinuous. In contrast, mural incorporation and obliterative changes were associated with alveolar structural remodeling. Electron microscopically, these lesions had bundles of normal collagen fibrils, small elastic fibers, and continuous epithelial basement membranes. These results indicate that: (i) intra-alveolar buds, that become intra-alveolar collagen globules, with an unusual extracellular matrix, do not contribute to alveolar structural remodelling; and (ii) areas of mural incorporation and obliterative changes have the usual type of extracellular matrix and are essential for alveolar structural remodelling.

Three-Dimensional Arrangement of Collagen and Elastin Fibers in the Human Urinary Bladder: A Scanning Electron Microscopic Study

To clarify the arrangements of collagen and elastin fibers of the urinary bladder, we examined 9 human (male, aged 42 to 72) urinary bladders by scanning electron microscopy with chemical digestion methods. The mucosal layer was divided into 3 portions according to the collagen arrangement: the superficial portion interwoven densely by collagen fibrils, the middle portion layered by flat bundles of collagen fibrils and the deep portion made of a loose network of twisted collagen bundles. In the muscular layer, the smooth muscle fascicles were firmly covered with collagen sheets, while each muscle cell in a fascicle was accommodated by a thin sheath of collagen fibrils. The serosal layer consists of wavy collagen bundles piled up in a sheet, which was intercalated by clusters of adipose cells. Elastic fibers were, on the other hand, sparse throughout the bladder wall, except for denser networks around the blood vessels and muscle fascicles and beneath the peritoneal mesothelium. The arrangements of these components were discussed in relation to the mechanical function and compliance of the urinary bladder.

Kupffer cell iron overload induces intercellular adhesion molecule-1 expression on hepatocytes in genetic hemochromatosis

The mechanisms underlying iron-induced liver fibrogenesis in patients with genetic hemochromatosis are poorly understood. We studied signs of Kupffer cell activation and inflammatory responses in liver biopsy specimens obtained from 15 patients with untreated and six patients with treated hemochromatosis. Immunohistochemistry was performed on 11 of the untreated and all treated patients. Three of the untreated patients (20%) had cirrhosis and eight (53%) had fibrosis. None had chronic active hepatitis (CAH). Immunohistochemistry indicated that 55% of the untreated patients had sparse intercellular adhesion molecule-1 (ICAM-1) expression by hepatocytes, and all of these had Kupffer cell iron overload. No ICAM-1 expression was seen by hepatocytes in treated patients or healthy controls. ICAM-1 was strongly expressed by hepatocytes from control patients with inflammatory liver disease. HLA-DR reactivity was seen on sinusoidal cells in all groups, but not on hepatocytes except for two of the control patients with CAH. Twenty-seven percent of the untreated hemochromatosis patients displayed moderate infiltration by CD3-positive lymphocytes. Electron microscopy of samples from untreated hemochromatosis patients showed hypertrophic Kupffer cells containing iron-rich remnants of phagocytosed hepatocytes. Fat-storing cells close to iron-laden hepatocytes contained multiple lipid droplets and adjacent collagen fibril bundles. Thus, in patients with untreated genetic hemochromatosis and Kupffer cell iron overload, hepatocytes occasionally express ICAM-1. In regions with heavy iron overload, Kupffer cell hypertrophy and transition of fat-storing cells are seen. Our findings indicate that release of factors from iron-loaded, activated Kupffer cells is of importance for the transformation of fat-storing cells and increased collagen deposition seen in genetic hemochromatosis.

Kupffer cell iron overload induces intercellular adhesion molecule-1 expression on hepatocytes in genetic hemochromatosis

The mechanisms underlying iron-induced liver fibrogenesis in patients with genetic hemochromatosis are poorly understood. We studied signs of Kupffer cell activation and inflammatory responses in liver biopsy specimens obtained from 15 patients with untreated and six patients with treated hemochromatosis. Immunohistochemistry was performed on 11 of the untreated and all treated patients. Three of the untreated patients (20%) had cirrhosis and eight (53%) had fibrosis. None had chronic active hepatitis (CAH). Immunohistochemistry indicated that 55% of the untreated patients had sparse intercellular adhesion molecule-1 (ICAM-1) expression by hepatocytes, and all of these had Kupffer cell iron overload. No ICAM-1 expression was seen by hepatocytes in treated patients or healthy controls. ICAM-1 was strongly expressed by hepatocytes from control patients with inflammatory liver disease. HLA-DR reactivity was seen on sinusoidal cells in all groups, but not on hepatocytes except for two of the control patients with CAH. Twenty-seven percent of the untreated hemochromatosis patients displayed moderate infiltration by CD3-positive lymphocytes. Electron microscopy of samples from untreated hemochromatosis patients showed hypertrophic Kupffer cells containing iron-rich remnants of phagocytosed hepatocytes. Fat-storing cells close to iron-laden hepatocytes contained multiple lipid droplets and adjacent collagen fibril bundles. Thus, in patients with untreated genetic hemochromatosis and Kupffer cell iron overload, hepatocytes occasionally express ICAM-1. In regions with heavy iron overload, Kupffer cell hypertrophy and transition of fat-storing cells are seen. Our findings indicate that release of factors from iron-loaded, activated Kupffer cells is of importance for the transformation of fat-storing cells and increased collagen deposition seen in genetic hemochromatosis.

The neuropathology of South American mummies.

Few studies of intracranial mummified brain tissue have been undertaken. This is because of the infrequency in which preserved human central nervous system tissue is encountered and the scarcity of available mummies from different parts of the world. This study undertook a systematic analysis of 15 naturally mummified human brains from 1000 B.C. to 1500 A.D. excavated from the deserts of northern Chile. Gross examination revealed relatively well-preserved dura mater, cerebral hemispheres, cerebellum, and spinal cord in several cases. Five cases showed evidence of intracranial disease. Three cases had evidence of external injury. One case revealed subarachnoid and one case revealed intracerebral hemorrhage. Samples of central nervous system tissues were taken for further analysis. The samples were rehydrated and processed for structural analysis by light and electron microscopy. Light microscopy of the brain parenchyma revealed an eosinophilic staining background with vascular structures but few cellular elements present. The dura mater demonstrated normal dural architecture consisting of collagen fibrils. Electron microscopy did not clearly demonstrate individual neurons or axonal processes. Bundles of collagen fibrils with typical periodicity were clearly seen in the dura mater. The examination of ancient human central nervous system tissues reveals normal and abnormal neuroanatomy.

Ectopic mineralized cartilage formation in human undifferentiated pancreatic adenocarcinoma explants grown in nude mice.

Mineralized as well as nonmineralized cartilage-like structures enclosing cells resembling chondrocytes were found in human-derived undifferentiated but not in poorly differentiated pancreatic adenocarcinoma explants grown in nude mice. The structures reacted with anti-mouse IgG but not with antibodies against human cytokeratin 19, indicating that the newly formed tissue was of mouse origin. High activity of alkaline phosphatase was found in cell layers surrounding the structures and in cells embedded in the matrix. The extracellular matrix was strongly positive after Sirius red staining, reacted with anti-collagen type II antibodies, and the presence of proteoglycans was demonstrated with Alcian blue staining and by metachromasia after Giemsa staining. Electron microscopic inspection revealed the presence of bundles of both thick collagenous fibrils with low levels of fine filamentous material and thin collagenous fibrils with high concentrations of filamentous components. The majority of both types of matrices was found to be partially or completely calcified. The mean area density of the cartilage-like structures in the undifferentiated tumors was 0.31%. The frequent formation of the cartilage-like structures in the rapidly growing undifferentiated explants and its absence in the slowly growing, more differentiated explants suggest that low oxygen tensions in combination with altered levels of growth factors, such as members of the transforming growth factor beta superfamily, create conditions that induce differentiation of fibroblasts to chondrocytes. It is concluded that these human tumors grown in nude mice can be used as an in vivo model to study ectopic formation of mineralized cartilage.

Detection of Proteoglycans in Human Posterior Capsule Opacification

Using cuprolinic blue staining, we histochemically examined the ultrastructural localization of proteoglycans in the fibrous-type human posterior capsule opacification. This capsule opacification contained proteoglycans that showed positive staining with cuprolinic blue. Many cuprolinic-blue-positive filaments were present within the bundles of collagen fibrils, basal lamina-like material and basal lamina of proliferated lens epithelial cells. Pretreatment with enzymes and nitrous acid revealed that the collagen fibrils contained chondroitin sulfate and dermatan sulfate, whereas the basal lamina-like material and basal lamina of epithelial cells contained heparan sulfate. These results suggest that proteoglycans are one of the components in posterior capsule opacification.

Scanning electron microscopic studies of reticular framework in the rat mesenteric lymph node

The reticular framework in the lymph node has in the past been studied mainly by light microscopy of silver-impregnated specimens. The aim of the present study is to understand three-dimensionally the ultrastructure and organization of the reticular framework better than before. The mesenteric lymph nodes of the rat were prepared either an alkali-water maceration method or a conventional method and were observed in a scanning electron microscope (SEM). The SEM study of alkali-water macerated tissues visualized directly the reticular fiber network in the lymph node. The reticular fibers consisted of thin bundles of collagen fibrils. They were continuous with the collagen fibrillar sheaths of blood vessels and lymphatic sinuses as well as with the fibrous capusule, thus acting as a skeleton of the lymph node. The arrangement of the reticulum was variable, depending on individual compartments. The SEM study of conventionally treated tissues, on the other hand, clarified the shape of reticular cells and their relationship with the reticular fibers. The sinus reticular cells connected with the sinus lining cells but separated from the parenchymal reticular cells, indicating that the former two originate from lymphatic endothelial cells. The parenchymal reticular cells varied in shape depending on their locations but essentially shared features with fibroblasts. The arrangements of the reticular fibers in the parenchyma were closely related to the associated reticular cells, showing the possibility that the reticular cells maintain the shape of the reticular framework suitable for each compartment of the lymph node.

Three-dimensional architecture of collagen fibrils in the mouse corpus cavernosum penis.

The three-dimensional architecture of the collagen fibrils in the corpus cavernosum penis of the mouse was studied after digestion of cellular and some extracellular elements by NaOH. The tunica albuginea was made up of undulating bundles of collagen fibrils which were arranged in outer longitudinal and inner circular layers. In both layers, several bundles were successively stacked to show the conspicuous lamellar arrangement. On the outer surface of the tunica, the collagen fibrils were interwoven like lacework. The trabecula appeared as a group of collagen bundles which extended from the inner circular layer of the tunica and showed a wavy arrangement. The grouped bundles in the trabecula were covered with a delicate sheath of collagen fibrils. These architectural features of the collagen fibrils appear to be related to fluctuations in the diameter and shape of the corpus during erection and detumescence.

Comparative study of the initial genesis of acellular and cellular cementum in rat molars

The initial genesis of acellular and cellular cementum was examined in rat molars by light and electron microscopy. Before root dentinogenesis, flattened dental follicle cells formed compartments by regularly arranged cellular processes which demarcated collagen fibril bundles oriented in parallel with the root long axis in both of the two kinds of cementum. After this stage, compartments disappeared from the dental follicle cells, which became elongated and polarized, with the cytoplasmic side facing toward the root surface in the acellular cementogenesis. Fibril bundles, oriented in parallel with the root long axis, decreased in number, and principal fibers appeared. Some principal fibers were attached on the first acellular cementum. Observations suggested that the fibril bundles, which had been oriented in parallel with the root long axis, were reoriented to merge into the principal fibers. In cellular cementogenesis, the dental follicle cells continued to hold the fibril bundles in cellular compartments. The regular processes were transformed into randomly oriented, finger-like processes. At the same time, fibers, which may be secreted from the finger-like processes, appeared around the preformed fibril bundles oriented in parallel with the root long axis. The different cellular behavior may result in the different fiber arrangement of acellular and cellular cementum.

An ultrastructural study of extracellular fibrillar components of developing mouse mandibular condyle with special reference to type VI collagen

The localization of type VI collagen was examined from birth to 8 weeks of age. Immunohistochemical staining with anti-type VI collagen antibody was strongly positive in the hypertrophic zone and moderately positive in the fibrous zone and the outer periphery of the proliferative zone, but negative in the inner area of the proliferative zone and mature zone. After ATP treatment, type VI collagen periodic fibrils with about 80-nm intervals were frequently observed but only in the fibrous zone. They occurred mainly in the superficial area of this zone, where striated collagen fibrils were sparse, while a few were noted in the inner area, where bundles of collagen fibrils were abundant. From these distributional differences of both components, a subzonation of the fibrous zone into superficial and inner areas is suggested. Moreover, with ATP treatment there were fewer type VI collagen periodic fibrils formed with increasing age (8 weeks). Testicular hyaluronidase digestion before ATP treatment facilitated the formation of periodic fibrils, in all the ages examined, in the intercellular space and around the fibroblastic cells. The interaction of type VI collagen with other components such as collagen fibrils, glycosaminoglycans or proteoglycans may play a part in maintaining the structural integrity of extracellular matrix in the mouse mandibular condyle.

Histological identification of osteocytes in the allegedly acellular bone of the sea breams Acanthopagrus australis, Pagrus auratus and Rhabdosargus sarba (Sparidae, Perciformes, Teleostei)

The bone of advanced teleost fishes such as those of the family Sparidae is said to lack osteocytes or to be acellular. Acellularity has been determined by apparent lack of osteocyte lacunae. This study questions the validity of this criterion. Scanning electron and light microscopy of paraffin and resin sections were used to show that the sides of sea bream mandibles consist of laminar parallel-fibred bone that we call tubular bone, because it contains tubules, and localised regions of Sharpey fibre bone. Osteocytes lie along the walls of tubules that also contain collagen fibril bundles (T-fibres), or in the lumens of tubules that do not contain T-fibres. We show that the osteocytes are derived from osteoblasts. The T-fibre system is different from other fibre systems that have been described. The tubules enclose wide T-fibres (lenticular in cross-section, maximum width about 8 microns) that taper at their ends and continue as thin T-fibres (round in cross-section, about 2 microns wide). The T-fibres originate in the periosteum. In mature tubular bone, spaces of increasing size develop around the osteocytes. Osteocytes are released from the bone matrix and become postosteocytes or bone-lining cells. Secondary bone lines the largest spaces. In Sharpey fibre bone, small osteocytes in small lacunae (about 2 microns wide) are found in columns parallel to the Sharpey fibres. Large osteocytes are found in large round spaces and are much larger than comparable osteocytes in lacunae in the bone of the salmon Salmo salar. We conclude that an absence of visible or conventional osteocyte lacunae does not mean that the cells themselves are absent. There are cells and two types of collagen fibre bundle in the tubules. The cells are osteocytes derived from osteoblasts, and these osteocytes apparently resorb bone with the result that large amounts of bone are destroyed. "Acellular" tubular and Sharpey fibre bone are types of cellular bone that differ from each other and from conventional cellular bone.

Corneal endothelial cell abnormalities in an early stage of the iridocorneal endothelial syndrome.

A corneal disc, obtained from a 52-year-old woman suffering from an early stage of the iridocorneal endothelial syndrome (ICE), was investigated by various morphological techniques to analyse the structural variations in the endothelial cells and to identify the collagen types within the abnormal layer of Descemet's membrane. Scanning electron microscopy of the posterior corneal surface revealed a mosaic of (a) flat hexagonal cells resembling irregular but normal endothelial cells, and (b) rounded hexagonal (ICE) cells with numerous surface microvilli. Degenerative changes were present in each cell type, but were more common in the flat hexagonal cells which contained intracytoplasmic spaces. By transmission electron microscopy the flat hexagonal cells exhibited many of the features of normal endothelial cells in terms of organelles and intercellular attachments, but lateral invaginations were absent. The ICE cells differed in that the apical surface was covered by microvilli and the cytoplasm contained tonofilaments, which were also observed by light microscopic immunocytochemical staining. Most commonly, intercellular attachments were rudimentary in both types of cell and intercellular spaces were dilated, but desmosomes were sometimes prominent in the ICE cells where interdigitations were pronounced. In some sectors, the basal surface of the ICE cells was indented by deposition of clumps of fibrillar collagenous material. An immunocytochemical study of the abnormal posterior deposits localised type IV collagen to the amorphous matrix and collagen types III and V, but not type I, to the collagen fibril bundles. Mononuclear inflammatory cells were identified between the ICE cells in the monolayer. The evidence suggests that some of the flat hexagonal cells were undergoing a degenerative change while others were transforming into ICE cells.

Expression of membrane alkaline phosphatase activity on gingival fibroblasts in chronic inflammatory periodontal disease.

We investigated the expression of membrane alkaline phosphatase (ALP) activity on fibroblasts in inflamed gingiva from 7 patients with adult periodontitis. ALP activity was ultrahistochemically detected by a cerium-based capture method. The degree of ALP activity was estimated by morphometric analysis of the percentage of the perimeter on which ALP reaction product was deposited. Fibroblasts in the non-inflammatory connective tissue were surrounded by bundles of collagen fibrils, and the majority of these fibroblasts showed ALP-negative or weakly ALP-positive reaction. By contrast, fibroblasts in the inflammatory connective tissue were either surrounded by a non-collagenous substance or in contact with inflammatory cells, and the majority of these fibroblasts showed a strong ALP-positive reaction. These results suggest that the expression of membrane ALP activity on gingival fibroblasts is induced by microenvironmental changes associated with the loss of contact between the cells and the extracellular collagenous matrix during inflammatory reactions.

Spatial Organization of the Collagen and Elastin Fibers of the Lung in the Japanese Monkeys, <i>Macaca fuscata</i>

The present study demonstrated the three-dimensional organization of the collagen and elastin fibers of the lung in the Japanese monkeys, Macaca fuscata as revealed by alkali-maceration/scanning electron microscope technique. The collagen fibers, which were bundles of collagen fibrils, were repeatedly divided and fused to form a network along the alveolar duct. The collagen fibers in the alveolar duct branched and entered the alveolar septa, where the fibers again repeated branching and fusing and formed thin networks. Thick collagen fibers around the alveolar mouths took significant degree of zig-zag or spiral courses. The elastin fibers were condensed around the alveolar mouths, but there were also elastin fibers between the adjacent alveolar mouths. Within the alveolar septa were course network of elastin fibers.

Structure and origin of the tooth pedicel (the so-called bone of attachment) and dental-ridge bone in the mandibles of the sea breams Acanthopagrus australis, Pagrus auratus and Rhabdosargus sarba (Sparidae, Perciformes, Teleostei).

Scanning electron and light microscopy were used to show that the pedicels of fish teeth (the so-called "bones of attachment") consist of three types of dentine that lie concentrically around a pulp cavity lined with typical odontoblasts with cytoplasmic processes in dentinal tubules. Circumpulpal canalar dentine forms on a thin layer of orthodentine that is encased in mantle dentine. Canalar dentine is a new name given to a dentine that is similar to vasodentine in canal arrangement, but not apparently in canal content. An inner series of wide, radial canals and an outer series of highly-branched thin canals of two diameters are inhabited by a population of cells, the osteodentocytes, and collagen fibril bundles. The flat, oval osteodentocytes appear to be quiescent cells, lying on the sides of the tubules and covered by a sheath. Plump, intensely metachromatic osteodentocytes appear to be more synthetically active. The canals and the osteodentocytes originate from blood capillaries enclosed in the predentine during dentinogenesis. New teeth begin within the small cavities present in spongy bone that were enlarged by multinucleated osteoclasts during tooth growth. Pedicel formation is initiated by the extension of the crown mantle dentine, forming the outer layer of the crimped ligament and outlining the future length and curvature of the pedicel. Central and inner ligament zones are subsequently formed as orthodentine is secreted in both crown and pedicel, and canalar dentine in the pedicel. Spongy bone osteogenesis begins during stage 1 of pedicel formation with the aggregation of osteoblasts and blood capillaries in the bone cavities and in the dermis between the pedicels. Loose fibrillar osteoid condenses into incomplete thin trabeculae bordered by intensely metachromatic osteoblasts. Osteoblasts become enclosed in the developing trabeculae that thicken to give mature spongy bone with osteocytes throughout. We conclude that the pedicels are the true bases of teeth, that the dental ridge is formed from pedicels and spongy bone, and that sea bream spongy bone is cellular. The term "bone of attachment" is inappropriate for the pedicel. It can be used for the spongy bone between the compact bone of the jaw and between adjacent pedicel.