RESUMO
This study compared the regeneration potentials of the hydroxyapatite (HA) and beta-tricalcium phosphate (ß-TCP) alone or in combination with a HA:TCP ratio of 30:70 in the critical-sized radial bone defects of rats. Bilateral 60 radial bone defects created were randomly divided into six equal groups (n=10 defects/group) including autograft, untreated or defect, chitosan-gelatin-platelet gel (CGP), CGP-HA, CGP-TCP and CGP-HA/TCP. The defects were evaluated by radiography, morphology, histopathology, histomorphometry, CT scan, scanning electron microscopy and biomechanical testing after eight weeks. Compared with the untreated and CGP-HA groups, the CGP and CGP-HA/TCP groups showed significantly higher new bone formation, bone volume, and mechanical properties. The CGP-HA and CGP-TCP scaffolds showed low biodegradability, whereas the CGP scaffolds were completely degraded. Osteoconductivity and osteoinductivity of the CGP and CGP-HA/TCP scaffolds were superior to the CGP-HA ones. The untreated and CGP-HA groups repaired mostly through fibrosis, while there were evidence of higher bone formation in the autograft, CGP and CGP-HA/TCP groups. In conclusion, addition of HA or ß-TCP alone into the CGP scaffolds impaired bone regeneration, while bone regeneration with the CGP and CGP-HA/TCP scaffolds was comparable with the autograft. Therefore, the CGP-HA/TCP scaffold can be a possible option to substitute the autologous bone grafting.
Assuntos
Plaquetas/química , Regeneração Óssea/efeitos dos fármacos , Fosfatos de Cálcio/química , Quitosana/química , Durapatita/química , Gelatina/química , Rádio (Anatomia)/efeitos dos fármacos , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Fenômenos Biomecânicos , Masculino , Teste de Materiais , Rádio (Anatomia)/fisiologia , Rádio (Anatomia)/ultraestrutura , Ratos , Ratos Sprague-Dawley , Ulna/efeitos dos fármacos , Ulna/fisiologia , Ulna/ultraestruturaRESUMO
Adipose-derived stem cells (ASCs) with multilineage potential can be induced into osteoblasts, adipocytes and chondrocytes. ASCs as seed cell are widely used in the field of tissue engineering, but most studies either use autologous cells as the source or an immunodeficient animal as the host. In our present study, we explored the feasibility of applying allogeneic ASCs and demineralized bone matrix (DBM) scaffolds for repairing tubular bone defects without using immunosuppressive therapy. Allogeneic ASCs were expanded and seeded on DBM scaffolds and induced to differentiate along the osteogenic lineage. Eight Sprague-Dawley (SD) rats were used in this study and bilateral critical-sized defects (8 mm) of the ulna were created and divided into two groups: with ASC-DBM constructs or DBM alone. The systemic immune response and the extent of bone healing were evaluated post-operatively. Twenty-four weeks after implantation, digital radiography (DR) testing showed that new bones had formed in the experimental group. By contrast, no bone tissue formation was observed in the control group. This study demonstrated that allogeneic ASCs could promote bone regeneration and repair tubular bone defects combined with DBM by histologically typical bone without systemic immune response.
Assuntos
Adipócitos/citologia , Regeneração Óssea , Células-Tronco/fisiologia , Engenharia Tecidual/métodos , Ulna/patologia , Adipócitos/ultraestrutura , Animais , Microscopia Eletrônica de Varredura , Osteogênese , Ratos Sprague-Dawley , Transplante de Células-Tronco/métodos , Células-Tronco/ultraestrutura , Alicerces Teciduais , Transplante Homólogo , Ulna/ultraestruturaRESUMO
Using in situ hybridization, we correlated the expression of mRNA for the parathyroid hormone/parathyroid hormone related peptide (PTH/PTHrP) receptor with bone formation and resorption in undecalcified serial sections of bones from growing rats. In addition we investigated the presence of biologically active receptors in the same locations using an in vivo autoradiographic technique. In the ulnae of growing rats, there are well defined zones of cortical bone formation and resorption. These contribute to the modeling drifts by which the bone achieves its adult shape. Forming surfaces incorporate fluorochrome labels, are lined with osteoid, and have a layer of cuboidal osteoblasts that have a high alkaline phosphatase activity. Resorbing surfaces have no fluorochrome incorporation, no osteoid, and are lined with resorbing cells with high tartrate-resistant acid phosphatase (TRAP) activity. PTH/PTHrP receptor mRNA was expressed predominantly on forming but not on resorbing bone surfaces and colocalized with sites of binding of radiolabeled PTH after intravenous injection. PTH/PTHrP mRNA expression on osteocytes was inconclusive but radiolabeled PTH bound to a proportion of osteocytes in all regions of the cortex although binding was not specifically related to areas of bone formation or resorption. These results suggest that in growing animals the actions of PTH or PTHrP are connected more with bone formation than resorption. Such a role may be linked to the ability of PTH to induce bone formation in adults but does not explain the actions of the hormone in regulating resorption. Binding of PTH to osteocytes increases the evidence for a physiological role for these cells.
Assuntos
Reabsorção Óssea/metabolismo , Osteoblastos/metabolismo , Osteócitos/metabolismo , Receptores de Hormônios Paratireóideos/biossíntese , Envelhecimento , Fosfatase Alcalina/metabolismo , Animais , Autorradiografia , Ligação Competitiva , Desenvolvimento Ósseo/fisiologia , Modelos Animais de Doenças , Imuno-Histoquímica , Hibridização In Situ , Osteoblastos/citologia , Osteoblastos/enzimologia , Osteócitos/citologia , Osteócitos/enzimologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptor Tipo 1 de Hormônio Paratireóideo , Receptores de Hormônios Paratireóideos/genética , Coloração e Rotulagem , Ulna/metabolismo , Ulna/fisiologia , Ulna/ultraestruturaRESUMO
Intracortical remodeling in the adult skeleton removes and replaces areas of compact bone that have sustained microdamage. Although studies have been performed in animal species in which there is an existing baseline of remodeling activity, laboratory rodents have been considered to have limited suitability as models for cortical bone turnover processes because of a lack of haversian remodeling activity. Supraphysiological cyclic axial loading of the ulna in vivo was used to induce bending with consequent fatigue and microdamage. Right ulnae of adult Sprague-Dawley rats were fatigue-loaded to a prefailure stopping point of 30% decrease in ulnae whole bone stiffness. Ten days after the first loading, left ulnae were fatigued in the same way. Ulnae were harvested immediately to allow comparison of the immediate response of the left ulna to the fatigue loads, and the biological response of the right leg to the fatigue challenge. Histomorphometry and confocal microscopy of basic fuchsin-stained bone sections were used to assess intracortical remodeling activity, microdamage, and osteocyte integrity. Bone microdamage (linear microcracks, as well as patches of diffuse basic fuchsin staining within the cortex) occurred in fatigue-loaded ulnar diaphyses. Ten days after fatigue loading, intracortical resorption was activated in ulnar cortices. Intracortical resorption occurred in preferential association with linear-type microcracks, with microcrack number density reduced almost 40% by 10 days after fatigue. Resorption spaces were also consistently observed within areas of the cortex in which no bone matrix damage could be detected. Confocal microscopy studies showed alterations of osteocyte and canalicular integrity around these resorption spaces. These studies reveal that: (1) rat bone undergoes intracortical remodeling in response to high levels of cyclic strain, which induce microdamage in the cortex; and (2) intracortical resorption is associated both with bone microdamage and with regions of altered osteocyte integrity. From these studies, we conclude that rats can initiate haversian remodeling in long bones in response to fatigue, and that osteocyte death or damage may provide one of the stimuli for this process.
Assuntos
Remodelação Óssea/fisiologia , Fraturas de Estresse/fisiopatologia , Ulna/fisiologia , Animais , Reabsorção Óssea , Microscopia Confocal , Ratos , Ratos Sprague-Dawley , Ulna/ultraestrutura , Suporte de CargaRESUMO
An in vivo study was designed to determine if non-invasive low intensity ultrasound could enhance bone growth into porous intramedullary implants. Fully porous intramedullary rods were implanted bilaterally into the ulnae of six dogs. In each dog, one ulna served as a control and the other was treated with 20 min of daily ultrasound stimulation for 6 consecutive weeks. Analysis of serial transverse sections indicated an average of 119% more bone growth into the ultrasound-treated implants compared with the contralateral controls (P < 0.001). In each of the 6 dogs, there was a significantly greater amount of bone ingrowth on the ultrasound-stimulated side. These data indicate a clear potential for externally applied ultrasound therapy to augment biological fixation.
Assuntos
Pinos Ortopédicos , Osseointegração/efeitos da radiação , Ulna/efeitos da radiação , Ulna/cirurgia , Ultrassom , Animais , Cães , Microscopia Eletrônica de Varredura , Porosidade , Radiografia , Ulna/diagnóstico por imagem , Ulna/ultraestruturaRESUMO
The interosseous membrane is a structure deep in the forearm that joins the radius and the ulna. It is made up of membranous and ligamentous regions. Two main ligamentous structures have been described: a prominent central fiber group, the "central band," and a smaller proximal fibrous band, the "oblique cord." Many authors believe that the central band plays a biomechanical role in the normal and fractured forearm and that it may function much like a ligament. The objective of this study was to determine the tensile properties of the central band. Eighteen fresh frozen forearms from cadavers (45-70 years of age, both sexes) were used. A fiber bundle of the central band was subjected to a uniaxial tensile test to failure in a materials testing machine, and its tensile properties were calculated. Stiffness, ultimate load, and energy absorbed to failure were expressed as a function of specimen width. The central band structure had a stiffness of 13.1 +/- 3.0 N/mm per mm width and an ultimate load of 56.6 +/- 15.1 N per mm width (mean +/- SD). The tissue of the central band displayed a modulus of 608.1 +/- 160.2 MPa, ultimate tensile strength of 45.1 +/- 10.3 MPa, and strain at failure of 9.0 +/- 2.0%. This study demonstrated that the central band is comprised of strong tissue. The material properties of the central band compare with those of patellar tendon: modulus is 120% and ultimate tensile strength is 84% that of patellar tendon. As a structure, the interosseous membrane is stiff and capable of bearing high loads. Although load distribution across the central band is unknown, a 1.7 cm wide, evenly loaded homogenous portion of the central band would possess a stiffness comparable with that of the anterior cruciate ligament. The results of this study provide a basis for future analyses of radioulnar stability and load transfer.
Assuntos
Antebraço/fisiologia , Idoso , Cadáver , Feminino , Humanos , Ligamentos/fisiologia , Masculino , Membranas/fisiologia , Pessoa de Meia-Idade , Rádio (Anatomia)/anatomia & histologia , Rádio (Anatomia)/fisiologia , Rádio (Anatomia)/ultraestrutura , Resistência à Tração , Ulna/anatomia & histologia , Ulna/fisiologia , Ulna/ultraestrutura , Suporte de Carga/fisiologiaRESUMO
There is evidence that the collagen microarchitecture of bone is influenced by mechanical stresses or strains. We hypothesized that peak functional strains correlate with the elastic anisotropy and collagen orientation of bone tissue and that the bone anisotropy might be changed by altering the strain patterns in canine radii for 12 months. We tested these hypotheses in studies using nine adult foxhounds. The baseline group (n = 3) had three rosette strain gauges placed around the midshaft of the radius, and strain distributions were measured during walking. The osteotomy group (n = 3) had 2 cm of the ulna surgically removed, and the sham group (n = 3) received a sham osteotomy. The osteotomy and sham groups were allowed free movement in cages with runs for 12 months, after which strain distributions were measured on the radii during walking. Bone-tissue anisotropy and collagen architecture were measured in radii from which the in vivo longitudinal strain patterns had been measured. The collagen birefringence patterns were measured with use of a circularly polarized light technique, and the elastic anisotropy of the bone, mineral, and collagen matrix was evaluated with a novel acoustic microscopy technique. Peak longitudinal strains in the radius correlated with the normalized longitudinal structure index (a polarized light measure of collagen birefringence) and the tissue anisotropy ratio. The average anisotropy ratio was 1.28+/-0.01 in the posterior (compressive) cortex and 1.43+/-0.01 in the anterior (tensile) cortex (these values are significantly different at p < 0.0001). The ulnar osteotomy changed the strain pattern on the radius, causing increased tensile strains in the medial cortex by more than 5-fold that were associated with a significant increase in the anisotropy ratio in the bone tissue. The longitudinal structure index was strongly correlated (r = 0.62, p < 0.005) with the anisotropy ratio of demineralized bone but was not correlated with that of deproteinized bone; this indicates that it reflects collagen fibril orientation in the bone matrix. These results indicate that mechanical strains affect both collagen and mineral microarchitecture in bone tissue, i.e., tensile strains are associated with increased tissue anisotropy and compressive strains, with decreased anisotropy.
Assuntos
Colágeno/fisiologia , Osteotomia , Rádio (Anatomia)/fisiologia , Animais , Anisotropia , Densidade Óssea/fisiologia , Remodelação Óssea/fisiologia , Colágeno/ultraestrutura , Cães , Elasticidade , Microscopia de Polarização , Rádio (Anatomia)/cirurgia , Rádio (Anatomia)/ultraestrutura , Estresse Mecânico , Resistência à Tração , Ulna/fisiologia , Ulna/cirurgia , Ulna/ultraestruturaRESUMO
It has been suggested that osteonal remodeling is triggered by bone microdamage. The validity of this theory rests on the assumption that loading within the physiological range will produce substantial microdamage with relatively few load cycles. The object of the first experiment was to determine threshold values required to consistently produce fatigue microdamage in vivo. The left forelimb of five groups of dogs, characterized by different strain levels and different numbers of load cycles, were loaded in three point bending. The number of microscopic fields which contained some microdamage was calculated as a percentage of the total number of fields. This experiment indicated that loads producing strains as low as 1500 microstrain on the radius and 1400 microstrain on the ulna for 10,000 cycles will produce significant bone microdamage. A second experiment was performed to verify this threshold and to determine whether microcracks are associated with the initiation of bone remodeling. Procedures in this experiment were the same as those in the first, except that all dogs were loaded in such a way as to produce strains on the radius of 1500 microstrain for 10,000 cycles, and the dogs were sacrificed 1-4 days after loading. The loaded limb demonstrated significantly more microdamage than the control limb (p = 0.03). Moreover, we observed 44 times as many microcracks in association with resorption spaces as expected by chance alone. These data support the hypothesis that fatigue microdamage is a significant factor in the initiation of intracortical bone remodeling.
Assuntos
Osso e Ossos/fisiologia , Animais , Desenvolvimento Ósseo , Reabsorção Óssea , Osso e Ossos/lesões , Cães , Masculino , Microscopia Eletrônica de Varredura , Rádio (Anatomia)/fisiologia , Rádio (Anatomia)/ultraestrutura , Estresse Mecânico , Ulna/lesões , Ulna/ultraestruturaRESUMO
An attempt to determine the locomotor activities of Mayulestes ferox (Borhyaenoidea) and Pucadelphys andinus (Didelphoidea) from the early Paleocene site of Tiupampa (Bolivia) is presented. The functional anatomy of the forelimbs of these South American marsupials is compared to that of some living didelphids: Caluromys philander, Micoureus demerarae, Marmosa murina, Didelphis marsupialis, Monodelphis brevicaudata and Metachirus nudicaudatus. Deductions from bone morphology to myology and locomotor behavior in the fossils are inferred from the comparisons with living forms. Some features of the postcranial skeleton, indicative of arboreal adaptations, are found in the extinct marsupials: anteriorly projected acromion, hemispherical head of the humerus, extended humeral lateral epicondylar ridge, medially protruding humeral entepicondyle, proximal ulnar posterior convexity, and deep flexor fossa on the medial side of the ulna. But other features are related to a more terrestrial pattern: the well-developed tubercles of the humeral head, the elongated olecranon process of the ulna, and the oval shape of the radial head. Mayulestes had clear arboreal abilities, but, as a predaceous mammal, probably hunted on the ground. Pucadelphys was less specialized, close to the living Monodelphis, a terrestrial insectivorous form with some skeletal features related to arboreal locomotion that are probably plesiomorphic for marsupials.
Assuntos
Membro Anterior/anatomia & histologia , Marsupiais/anatomia & histologia , Gambás/anatomia & histologia , Adaptação Fisiológica , Animais , Evolução Biológica , Fenômenos Biomecânicos , Membro Anterior/fisiologia , Fósseis , Úmero/anatomia & histologia , Úmero/fisiologia , Locomoção , Marsupiais/fisiologia , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/fisiologia , Gambás/fisiologia , Rádio (Anatomia)/anatomia & histologia , Rádio (Anatomia)/fisiologia , Escápula/anatomia & histologia , Escápula/fisiologia , Articulação do Ombro/anatomia & histologia , Articulação do Ombro/fisiologia , Ulna/fisiologia , Ulna/ultraestruturaRESUMO
In the present study, we tested the hypothesis that tension and bending, rather than compression alone, determine the functional adaptation of subchondral bone in incongruous joints. We investigated whether tensile stresses in the subchondral bone of the humero-ulnar articulation are affected by the direction of muscle and joint forces, and whether the tensile stresses are large enough to cause microstructural adaptation, specifically a preferential alignment of the trabeculae and the subchondral collagen fibres. Using a previously validated finite element model of the human humero-ulnar joint, we calculated the contact pressure, the principal compressive and tensile stresses, and the strain energy density in the subchondral bone for various flexion angles. A bicentric (ventro-dorsal) pressure distribution was found in the joint at 30 degrees to 120 degrees of flexion, with contact pressures of up to between 2.5 and 3 MPa in the ventral and dorsal aspects of the ulnar joint surface, but less than 0.5 MPa in the centre. The principal tensile stress in the subchondral bone of the trochlear notch quantitatively exceeded the principal compressive stress at low flexion angles (maximum 8.2 MPa), and the distribution of subchondral strain energy density differed substantially from that of the contact stress (r=-0.72 at 30 degrees and r=+0.58 at 90 degrees of flexion). No important tensile stress was computed in the trochlea humeri. On contact radiography, we found sagittally orientated subarticular trabeculae in the notch, running tangential to the surface. Furthermore, we observed sagittally orientated split lines in the subchondral bone of the notch of 20 cadaver joints, suggesting a ventro-dorsal orientation of the collagen fibres. The trochlea humeri, on the other hand, did not show a preferential direction of the subchondral split lines, these findings confirming the predictions of tensile stresses in the model. We conclude that, due to the important contribution of tension to subchondral bone stress, the distribution of subchondral density cannot be directly employed for assessing the long term distribution of joint pressure at the cartilage surface. The magnitude of the tensional stress varies considerably with the direction of the muscle and joint forces, and it appears large enough to cause functional adaptation of the subchondral bone on a microstructural level.
Assuntos
Adaptação Fisiológica/fisiologia , Osso e Ossos/fisiologia , Articulação do Cotovelo/fisiologia , Densidade Óssea , Osso e Ossos/ultraestrutura , Força Compressiva/fisiologia , Simulação por Computador , Articulação do Cotovelo/ultraestrutura , Análise de Elementos Finitos , Humanos , Úmero/fisiologia , Úmero/ultraestrutura , Modelos Biológicos , Pressão , Estresse Mecânico , Resistência à Tração , Ulna/fisiologia , Ulna/ultraestrutura , Suporte de CargaRESUMO
Twenty-five ununited coronoid processes (UCP) and 24 osteochondritis dissecans (OD) flaps were examined by light and transmission electron microscopy. Chondrocytes showed degenerative changes but remained viable and continued to secrete matrix components, even though the organisation of the matrix was altered. Differences in the histological and ultrastructural appearance of the two lesions tend to suggest that they are two separate disease entities, although they may occur together in the same joint. It is hypothesised that OD results from incorrect cartilage maturation and endochondral ossification. The aetiology of UCP is unclear but there is a possibility of its being a subchondral fracture, with an ineffective fibrous repair in some cases.
Assuntos
Cartilagem Articular/patologia , Doenças do Cão/etiologia , Osteocondrite Dissecante/veterinária , Ulna/patologia , Animais , Cartilagem Articular/ultraestrutura , Doenças do Cão/patologia , Cães , Membro Anterior , Microscopia Eletrônica , Osteocondrite Dissecante/etiologia , Osteocondrite Dissecante/patologia , Ulna/ultraestruturaRESUMO
The elbows of 13 puppy cadavers were dissected, samples were taken for light and electron microscopy, and the thickness of the articular cartilage of the distal humerus and proximal ulna was measured. Throughout post natal development differences were found in the arrangement of the growth plate and articular chondrocytes. At birth, the articular surface had remnants of a fibrous limiting membrane that was continuous with the perichondrium, a finding not previously recorded in dogs. Orientation of the collagen fibrils within the matrix of the articular cartilage was initially lacking but became established by three weeks. In the humerus cartilage canals were present up to 12 weeks old. The articular cartilage of the humeral condyle varied in thickness across the joint surface, being thicker on the medial than on the lateral side; it was also thicker at the apex of the medial coronoid process. These regions of thick cartilage correspond with the sites where cartilage defects arise in elbow osteochondrosis. No histological evidence was found that the medial cornoid process of the ulna is a separate centre of ossification.
Assuntos
Animais Recém-Nascidos/crescimento & desenvolvimento , Cães/crescimento & desenvolvimento , Membro Anterior/crescimento & desenvolvimento , Animais , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/ultraestrutura , Membro Anterior/ultraestrutura , Úmero/crescimento & desenvolvimento , Úmero/ultraestrutura , Articulações/crescimento & desenvolvimento , Articulações/ultraestrutura , Microscopia Eletrônica , Osteogênese , Ulna/crescimento & desenvolvimento , Ulna/ultraestruturaRESUMO
Transport of fluorescent probes between 300 and 2,000,000 Da was studied in mechanically loaded and unloaded ulnae of skeletally mature rats to characterize the permeability of the pericellular space of the lacunocanalicular system (LCS), and the microporosity of the bony matrix. The mineral matrix porosity allowed for penetration of the 300 Da probe but impeded transport of larger probes. The pericellular space of the LCS was permeable up to 10 kDa; above 10 kDa, diffusion was ineffective for transport through the pericellular space. Convective transport via load-induced fluid flow increased penetration of all probes up to 70 kDa. Above this threshold, probes were excluded from bone, both with and without loading. This exploratory study suggests that bone acts as a molecular sieve and that mechanical loading modulates transport of solutes through the pericellular space that links osteocytes deep within the tissue to the blood supply and to osteoblasts and osteoclasts on bone forming and resorbing surfaces. This provides support for the postulate of transport modulated bone remodeling in which osteocytes are influenced by and modulate the local permeability of their surroundings as a means for survival (Knothe Tate et al. 1998, [28]) and has profound implications for osteocyte viability and intercellular communication in bone.
Assuntos
Osso e Ossos/fisiologia , Mecanotransdução Celular/fisiologia , Animais , Transporte Biológico Ativo/fisiologia , Osso e Ossos/ultraestrutura , Líquido Extracelular/fisiologia , Feminino , Técnicas de Sonda Molecular , Osteócitos/fisiologia , Permeabilidade , Ratos , Ratos Sprague-Dawley , Reologia , Estresse Mecânico , Ulna/fisiologia , Ulna/ultraestrutura , Suporte de Carga/fisiologiaRESUMO
Chondrocytes from the distal ulnar physis of 1- and 5-month-old boars were studied by transmission electron microscopy. Among reserve, proliferating, and early hypertrophic chondrocytes, 2 distinct cell types (dark cells and light cells) were recognized, and in the distal proliferative and early hypertrophic regions, dark cells seemed to be involved in matrix vesicle production. Late hypertrophic light cells seemed to follow 2 lines of maturation. One pathway resulted in cells with a stellate shape, typical of hypertrophic cells in several species other than swine. A 2nd pathway resulted in large oval cells with water-clear cytoplasm. These cells appeared to become encased in a lamina of fine fibrils interspersed with irregularly shaped electron-dense material. The functional significance of these 2 pathways remains unclear.
Assuntos
Cartilagem/ultraestrutura , Epífises/ultraestrutura , Suínos/anatomia & histologia , Animais , Cartilagem/crescimento & desenvolvimento , Epífises/crescimento & desenvolvimento , Hipertrofia , Masculino , Microscopia Eletrônica , Suínos/crescimento & desenvolvimento , Ulna/crescimento & desenvolvimento , Ulna/ultraestruturaRESUMO
Mechanical environment is one of the regulating factors involved in the process of longitudinal bone growth. Non-physiological compressive loading can lead to infantile and juvenile musculoskeletal deformities particularly during growth spurt. We hypothesized that tissue mechanical behavior in sub-regions (reserve, proliferative and hypertrophic zones) of the growth plate is related to its collagen and proteoglycan content as well as its collagen fiber orientation. To characterize the strain distribution through growth plate thickness and to evaluate biochemical content and collagen fiber organization of the three histological zones of growth plate tissue. Distal ulnar growth plate samples (N = 29) from 4-week old pigs were analyzed histologically for collagen fiber organization (N = 7) or average zonal thickness (N = 8), or trimmed into the three average zones, based on the estimated thickness of each histological zone, for biochemical analysis of water, collagen and glycosaminoglycan content (N = 7). Other samples (N = 7) were tested in semi-confined compression under 10% compressive strain. Digital images of the fluorescently labeled nuclei were concomitantly acquired by confocal microscopy before loading and after tissue relaxation. Strain fields were subsequently calculated using a custom-designed 2D digital image correlation algorithm. Depth-dependent compressive strain patterns and collagen content were observed. The proliferative and hypertrophic zone developed the highest axial and transverse strains, respectively, under compression compared to the reserve zone, in which the lowest axial and transverse strains arose. The collagen content per wet mass was significantly lower in the proliferative and hypertrophic zones compared to the reserve zone, and all three zones had similar glycosaminoglycan and water content.Polarized light microscopy showed that collagen fibers were mainly organized horizontally in the reserve zone and vertically aligned with the growth direction in the proliferative and hypertrophic zones. Higher strains were developed in growth plate areas (proliferative and hypertrophic) composed of lower collagen content and of vertical collagen fiber organization. The stiffer reserve zone, with its higher collagen content and collagen fibers oriented to restrain lateral expansion under compression, could play a greater role of mechanical support compared to the proliferative and hypertrophic zones, which could be more susceptible to be involved in an abnormal growth process.
Assuntos
Matriz Extracelular/fisiologia , Colágenos Fibrilares/fisiologia , Lâmina de Crescimento/fisiologia , Mecanotransdução Celular/fisiologia , Modelos Biológicos , Ulna/fisiologia , Animais , Força Compressiva/fisiologia , Simulação por Computador , Módulo de Elasticidade/fisiologia , Matriz Extracelular/ultraestrutura , Colágenos Fibrilares/ultraestrutura , Lâmina de Crescimento/ultraestrutura , Técnicas In Vitro , Estresse Mecânico , Suínos , Ulna/ultraestruturaRESUMO
The bisphosphonate zoledronic acid chemically and physically was bound to hydroxyapatite-coated porous tantalum implants. The zoledronic acid elution characteristics in saline were determined as a function of time and the in vivo effects of elution were quantified at 12 weeks in a canine ulnar implant model. Intramedullary implants surgically were implanted bilaterally into the ulnae of a control group of five dogs and a zoledronic acid-dosed (0.05 mg zoledronic acid) group of four dogs. Computerized image analysis of undecalcified histologic sections was used to quantify the amount of peri-implant bone within the intramedullary canal, the percentage of available pore space filled with new bone, and the number and size of the individual bone islands within the implant pores. The data were analyzed using a hierarchical analysis of variance with 95% confidence intervals. The peri-implant bone occupied a mean of 13.8% of the canal space in controls and 32.2% of the canal space in zoledronic acid-dosed dogs, a relative difference of 134% (2.34-fold) that was significant. The mean extent of bone ingrowth was 12.5% for the control implants and 19.8% for the zoledronic acid-dosed dogs, a relative difference of 58% that was statistically significant. Individual islands of new bone formation with the implant pores were similar in number in both implant groups but were 71% larger on average in the ZA-dosed group. We are the first authors to show that local elution of a bisphosphonate can cause substantial bone augmentation around and within porous orthopaedic implants. The concept represents a potential tool for restoration of bone stock and enhancement of implant fixation in primary and revision cementless joint arthroplasty surgeries in the face of compromised or deficient bone.
Assuntos
Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Imidazóis/farmacologia , Osseointegração/efeitos dos fármacos , Próteses e Implantes , Ulna/cirurgia , Animais , Materiais Revestidos Biocompatíveis/farmacologia , Cães , Sistemas de Liberação de Medicamentos , Durapatita/farmacologia , Microscopia Eletrônica de Varredura , Modelos Animais , Radiografia , Tantálio , Ulna/diagnóstico por imagem , Ulna/ultraestrutura , Ácido ZoledrônicoRESUMO
A scanning electron microscopic (SEM) cast method that has been utilized to examine the internal structure of dentine was modified to examine canalicular communications in the cortices of a human femur and ulna. Although some preparations in which all of the matrix was removed were examined, etched preparations were found to be the most informative. Casts of lacunae and canaliculi along with the underlying matrix could be visualized in these preparations. In the femur, whose cortex exhibited a typical lamellar pattern, canalicular communication was seen between first and second generation osteons and occasionally between osteons and interstitial regions. The interstitial regions in the ulna appeared to be primarily woven rather than lamellar bone. Extensive communication between the outermost lacunae of osteons and interstitial regions was observed in the ulna.
Assuntos
Comunicação Celular , Fêmur/citologia , Ulna/citologia , Fêmur/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura/métodos , Modelos Anatômicos , Ulna/ultraestruturaRESUMO
de Margerie hypothesized that preferred orientations of primary vascular canals in avian primary cortical bone mediate important mechanical adaptations. Specifically, bones that receive habitual compression, tension or bending stresses typically have cortices with a low laminarity index (LI) (i.e. relatively lower cross-sectional areas of circularly (C) orientated primary vascular canals, and relatively higher areas of canals with radial (R), oblique (O) or longitudinal (L) orientations. By contrast, bones subject to habitual torsion have a high LI (i.e. relatively higher C-orientated canal area) [LI, based on percentage vascular canal area, = C/(C + R + O + L)]. Regional variations in predominant collagen fibre orientation (CFO) may be the adaptive characteristic mediated by LI. Using turkey ulnae, we tested the hypothesis that site-specific variations in predominant CFO and LI are strongly correlated. Mid-diaphyseal cross-sections (100 +/- 5 micro m) from subadult and adult bones were evaluated for CFO and LI using circularly polarized light images of cortical octants. Results showing significant differences between mean LI of subadult (40.0% +/- 10.7%) and adult (50.9% +/- 10.4%) (P < 0.01) bones suggest that adult bones experience more prevalent/predominant torsion. Alternatively, this relationship may reflect differences in growth rates. High positive correlations between LI and predominant CFO (subadults: r = 0.735; adults: r = 0.866; P < 0.001) suggest that primary bone can exhibit potentially adaptive material variations that are independent of secondary osteon formation.
Assuntos
Colágeno , Perus/fisiologia , Ulna/irrigação sanguínea , Adaptação Fisiológica/fisiologia , Animais , Microscopia de Polarização , Estresse Mecânico , Perus/anatomia & histologia , Ulna/anatomia & histologia , Ulna/ultraestruturaRESUMO
Cultured chondrocytes were bound to hydroxyapatite (HA) and implanted in surgically created bone defects in the ulna of the rabbit. Chondrocytes were obtained from the iliac crest cartilage of the rabbit and were cultured on the HA block for two weeks before implantation. Scanning electron micrographs showed that pores of the HA surface and the inside of the block were filled with cultured cells. After cultivation for two weeks, the expression of chondrocyte-specific characteristics was confirmed in these cells by an implanted diffusion of chondrocyte-specific characteristics was confirmed in these cells by an implanted diffusion chamber experiment. The HA without chondrocytes served as a control. At two weeks postimplantation, proliferating chondrocytes were observed on the surface and in the inside pores of the HA-chondrocyte blocks but not the HA controls. At four and six weeks postimplantation, in the central region of the implants, there was a significant increase in the amount of bone formation in the HA with cultured chondrocytes. At 13 weeks post-implantation, the implants were partially resorbed and completely enveloped in lamellar bone, including bone marrow.
Assuntos
Regeneração Óssea , Cartilagem/citologia , Hidroxiapatitas , Ulna/citologia , Animais , Cartilagem/transplante , Células Cultivadas , Durapatita , Microscopia Eletrônica de Varredura , Osteogênese , Próteses e Implantes , Coelhos , Radiografia , Ulna/diagnóstico por imagem , Ulna/ultraestruturaRESUMO
The scanning electric microscopy (SEM). X-ray energy graph and Ca, P contents analysis were adopted in studying the mechanism and superiority of two different composite artificial bones (HA-bBMP-Co and HA-Co) to repair segmental bone defect in rabbits. The results indicated that HA-bBMP-Co had a double effects of bone induction and bone conduction. No difference of P content in two weeks after reconstructive surgery was found, but in other periods, there was an obvious difference in Ca and P content between HA-bBMP-Co group and HA-Co group (P < 0.05).