M. Raspanti

Human Achilles Tendon: Morphological and Morphometric Variations as a Function of Age

Aging of human Achilles tendon results in changes in both cellular and fibrous components. Cells flatten and become less numerous. Their thin and long cytoplasmatic projections tend to shorten and diminish in number. Tendon fibers lose their typical undulating appearance and become quite straight. Collagen fibril diameter, small and uniform in the neonatal period, becomes large and extremely variable from adolescence onwards. Age related morphometric changes include a decrease in the average, maximum diameter and density of collagen fibrils and an increase of fibril concentration. In our opinion these morphological and morphometric variations are strictly related to functional requirements.

Detachment of titanium and fluorohydroxyapatite particles in unloaded endosseous implants

The shape, surface composition and morphology of orthopaedic and endosseous dental titanium implants are key factors to achieve post-surgical and long-term mechanical stability and enhance implant osteointegration. In this study a comparison was made between 12 titanium screws, plasma-spray-coated with titanium powders (TPS), and 12 screws with an additional coating of fluorohydroxyapatite (FHA-Ti). Screws were implanted in the femoral and tibial diaphyses of two mongrel sheep and removed with peri-implant tissues 12 weeks after surgery. The vibrational spectroscopic, ultrastructural and morphological analyses showed good osteointegration for both types of implants in host cortical bone. The portion of the FHA-Ti implants in contact with the medullary canal showed a wider area of newly formed peri-implant bone than that of the TPS implants. Morphological and EDAX analyses demonstrated the presence of small titanium debris in the bone medullary spaces near the TPS surface, presumably due to the friction between the host bone and the implant during insertion. Few traces of titanium were detected around FHA-Ti implants, even if smaller FHA debris were present. The present findings suggest that the FHA coating may act as a barrier against the detachment of titanium debris stored in the medullary spaces near the implant surface.

Ultrastructure of heat-deproteinated compact bone

Heat-deproteinated bone (calcined bone) is a natural candidate for an osteoreproductive biomaterial. Ultrastructural investigation has pointed out an important aspect of the preparation technique. Treatment of bone at temperatures exceeding 500 degrees C causes complete disruption of the tissue architecture and the reorganization of the mineral phase into tightly packed, dense crystals. At 500 degrees C or less the structure and distribution of the mineral phase remain unaffected, so that cross-banded shadows of collagen fibrils are still readily observable, although collagen is no longer present in the samples. By its excellent structural preservation and natural porosity, low-temperature calcined bone seems to be a promising alternative for osteoreproduction.

Low-temperature heat-deproteinated compact bone to heal large bone defects.

The potential of low-temperature (400 degrees C), heat-treated bone matrix in osteorepair has been evaluated in vivo by implantation into defects artificially created in rodent tibia. Histological and ultrastructural analysis of the bone--implant interface has been carried out on samples obtained at 1 to 6 weeks from operation. The obtained data showed that calcined bone is well tolerated and does not cause acute or chronic inflammatory reactions. Osteoid tissue, tightly adhered to the implant, appears within 2 weeks of the operation, while after 6 weeks newly formed bone surrounds and infiltrates the implant. Of greater note, the detection of good adhesion between bone and implant ultrastructurally is demonstrated by the absence of fibrillar connective tissue at the interface. For these reasons, our preliminary observations suggest that low-temperature calcined bone (biological apatite or heat-deproteinated bone) may have a rightful place among the osteointegrators.

Collagen Fibril Patterns in Compact Bone: Preliminary Ultrastructural Observations

A comparative study of the Haversian architecture was carried out on compact bone derived from the anterior and posterior edges of the diaphysis of horse radius, regions which have different mechanical requirements in vivo. Samples were heat-deproteinated prior to SEM analysis, a treatment which effectively removes cells and vascular structures as well as exposing large areas of the mineralization front along the walls of the haversian canals. Bone subject to tensile stress revealed a prevalent alignment of its collagen fibrils in the stress direction, and the vast majority of its osteons were composed of fibrils running almost parallel and crossing at very acute angles. Bone subject to compressive forces showed either an orthogonal alternation of collagen lamellae or a multidirectional arrangement corresponding to the twisted plywood pattern described by other authors. Our observations substantiate both the classical model of the osteon and the twisted plywood concept, and suggest that osteon ultrastructure is modulated according to biomechanical requirements.

Structural and functional macrophages alterations by ceramics of different composition.

Biomaterials may initiate several and complex biological reactions in host tissues, and the cell-biomaterial interactions can determine the release of mediators including monocytes and lymphocytes chemotactic factors. The present work was aimed to investigate in vitro the macrophage biological reactions of a natural apatite obtained by heat treatment at 400 degrees C of bovine bone, compared to other ceramics usually used for dental and orthopedic applications, using synthetic apatite and three types of alumina as controls. Particles chemotactic activity and powders oxidative burst evidenced no monocyte macrophages sensitivity reaction for natural and synthetic hydroxyapatite powders at great granulometry (> 50 microm); data were confirmed by ultrastructural observations; electron microscopy analysis showed macrophages with the features of healthy cells in the presence of both natural and synthetic apatites while macrophages grown in the presence of alumina seemed to be negatively affected. In conclusion, among all ceramics tested, natural apatite displayed a good compatibility with living cells, being better tolerated than synthetic hydroxyapatite which in turn is better tolerated than alumina.

Particular Structure of the Anterior Third of the Human True Vocal Cord

The histological aspects of the true vocal cord mucosa change in the anterior third compared with the posterior two thirds. The anterior third is characterized by an epithelium where the ridges, marked in the posterior two thirds, are very slight or even absent. The underlying basement membrane, which is thin in the posterior two thirds, here appears particularly thick. At the ultrastructural level in this area, beneath a normally thickened basal lamina, a thick layer of finely granulated electron-dense material, interspersed with thin and randomly scattered collagen fibrils and proteoglycan filaments, is detectable. Beneath this thickened basement membrane, a layer of small undulated collagen fibril bundles with very numerous interspersed oxytalan fibres is found. The collagen fibrils, small in diameter (30-40 nm), seem to continue with the collagen fibrils of the basement membrane. In this layer numerous blood vessels with a very thick, delaminated basement membrane are also observed. The underlying area is characterized by the vocal cord ligament, composed by large compact collagen fibril bundles with interspersed elastic fibres. The particular features of the thick basement membrane, the thick-walled and delaminated vessels and the modular distribution of the elastic system together may well form the basic structure enabling the functional integration of the vocal ligament into the overlying mucosa and the underlying vocal muscle.

Ultrastructural aspects of freeze-fractured and etched elastin

The ultrastructural organization of fresh and purified elastin from beef ligamentum nuchae was studied by means of the freeze-etching technique. Both fresh and purified elastin showed a regular three-dimensional network of filaments which seemed to be composed of a sequence of globular subunities. There were also areas, along the regular network, in which ridges of various lengths, packed with perpendicular side filaments, were visible. In replicas of deep-etched and rotary-shadowed specimens, a thicker and more defined three-dimensional network was observable. A great variability in appearance among the globular subunits of the filaments was noticed which was at least partially due to the etching treatment. By means of computerized simulation of replicas of various hypothetically collapsed globular structures, we obtained patterns which were superimposable on those obtained in the replicas of the specimens analyzed. It is thus assumed that each globular subunit of the filament, being subjected to collapsing, has a less dense central core.