RESUMEN
The aim of the study was to assess the normal cranial suture and bone diploae ultrastructural morphology. Two types of sutures from different specimens were collected. The micro-CT scanning provided a three-dimensional view of the sutures at a microscopic level thus allowing the evaluation of the development stage and a rapid analysis evaluation of bone and diploae morphology. In the meantime, the micro-CT is able to generate more slices than the normal histology preserving the analyzed specimens and became one of the most powerful tools in the craniofacial area. The micro-CT analysis generated structure-orientated slices that in conjunction with the histological sections provide a high quality quantitative analysis of all cranial sutures and of the cranial bones diploae.
Asunto(s)
Huesos/anatomía & histología , Suturas Craneales/anatomía & histología , Adolescente , Adulto , Huesos/diagnóstico por imagen , Huesos/ultraestructura , Suturas Craneales/diagnóstico por imagen , Suturas Craneales/ultraestructura , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Microtomografía por Rayos XRESUMEN
In addition to acting as a growth site, sutures in the facial skeleton are important for distributing mechanical forces during mastication. In the present study, the extent of fusion of a facial suture is assessed in two samples of adult bushbabies (Galago moholi and Otolemur garnettii). Microanatomical techniques were used to determine the loci of osseous bridges across the nasomaxillary suture (NMS). Histological sections containing sutures with osseous bridging were rated as "fused." One of the specimens was studied using micro-computed tomography before paraffin embedding and serial sectioning. At all ages, O. garnettii shows more advanced fusion of the NMS than G. moholi. The youngest O. garnettii shows multiple foci of fusion of the NMS; however, 13% of the posterior most suture is patent. Throughout the NMS of this animal, sutural fusion is isolated to one or two small osseous bridges, typically of woven bone. These bridges are most often on the external (superficial) surface of the suture, but in numerous sections the site of fusion occurs deep to an external notch. In G. moholi, the youngest adults studied showed little or no fusion across the NMS. However, the nasal and maxillary bones were indirectly tethered at some levels by other bones that were fused to both nasal and maxillary bones. These results indicate that microanatomical evidence is required to fully assess the extent of fusion of facial sutures. These findings also support previous observations of differing magnitude of suture fusion between the two species.
Asunto(s)
Suturas Craneales/ultraestructura , Galago/anatomía & histología , Maxilar/ultraestructura , Hueso Nasal/ultraestructura , Primates/anatomía & histología , Animales , Suturas Craneales/diagnóstico por imagen , Maxilar/diagnóstico por imagen , Desarrollo Musculoesquelético , Hueso Nasal/diagnóstico por imagen , Especificidad de la Especie , Tomografía Computarizada por Rayos XRESUMEN
AIM: Determining age at the time of death is a difficult problem in forensic practice. The study of the vascularization of cranial sutures is an original approach, which may mark the process of synostosis associated with aging. Counting sections of blood vessels on a histological section of cranial suture, raises however a number of practical problems related to the quality of the preparation and to the method of quantification. The purpose of this study is to illustrate the potential contribution of an automatic analysis of virtual slides to overcome these problems. MATERIALS AND METHODS: The proposed method of analysis is illustrated from three samples of frontosphenoidal suture whose vessels were immunostained after decalcification of bone tissue. A high resolution image (x 20 objective and microscopic resolution of 0.5 microm) of each microscopic preparation was acquired through a microscopic scanning device. The automatic image analysis protocol takes advantage of a processing of virtual slides at two resolutions. RESULTS: The chosen strategy ensures the identification of the specific area of interest, the enumeration of blood vessel sections on the whole preparation and the visual control of the detected structures. CONCLUSION: The quantitative estimate of the vascularization of a large structure, such as a cranial suture, can benefit from scanning and fully automatic processing of virtual slides. The automatic analysis requires however an optimal preparation of tissues; decalcification and immunohistochemical staining must be done with great care.
Asunto(s)
Suturas Craneales/patología , Procesamiento de Imagen Asistido por Computador/métodos , Neovascularización Patológica/patología , Automatización , Calcinosis , Suturas Craneales/ultraestructura , Humanos , Procesamiento de Imagen Asistido por Computador/instrumentación , Análisis de Regresión , Cráneo/patología , Interfaz Usuario-ComputadorRESUMEN
Although recent studies indicate that regional dura mater influences the fate of the overlying cranial suture, little is known about the assembly of extracellular matrix (ECM) molecules within the patent and fusing murine cranial suture complexes. Confocal laser scanning microscopy was used to study ECM assembly within patent and fusing cranial suture complexes. Coronal sections (20 microm thick) of patent sagittal (SAG) and fusing posterior frontal (PF) sutures from postnatal 8-, 14-, and 18-day-old Sprague-Dawley rats were scanned in 0.5-microm increments, and images were collected consecutively to create a z-series for three-dimensional reconstruction. Spatial and temporal collagen arrangements were compared between SAG and PF sutures by measuring interfiber distance, fiber thickness, and total collagen surface area at each time point. We demonstrate that on day 8 (before the onset of suture fusion), collagen bundles are randomly arranged in both the SAG and PF sutures. By day 14 (midfusion period), there was a statistically significant reduction in total collagen surface area (80.5% versus 67.4%; P < 0.05) as the collagen bundles were organized into orthogonal lattices along the anterior and endocranial margins of the PF suture. Furthermore, new bone matrix deposition was observed along the edges of these organized collagen bundles. In contrast, collagen within the SAG suture remained randomly arranged and unossified. By day 18 (late fusion period), the PF suture was completely fused except for the posterior-ectocranial portion. This patent section of the PF suture contained a highly organized mineralizing orthogonal collagen lattice. The total collagen surface area in the day-18 PF suture continued to decline compared with the day-8 PF suture (80.5% versus 55.6%; P < 0.05). In the day-18 SAG suture, the collagen bundles remained randomly arranged, and the total surface area did not change. The same analysis was performed in a human pathologic fusing and patent suture. Similar results were observed. The total collagen surface area significantly decreased in the pathologic fusing human suture compared with the patent suture (92.8% versus 60.6%; P < 0.05). Moreover, the pathologically fusing suture contained a highly organized mineralizing orthogonal collagen lattice. This is the first analysis of collagen patterns in patent and fusing cranial sutures.
Asunto(s)
Colágeno/ultraestructura , Suturas Craneales/ultraestructura , Animales , Matriz Ósea/ultraestructura , Craneosinostosis/patología , Colágenos Fibrilares/ultraestructura , Hueso Frontal/ultraestructura , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Lactante , Masculino , Mesodermo/ultraestructura , Microscopía Confocal , Osteogénesis/fisiología , Hueso Parietal/ultraestructura , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: Neonates with Down syndrome are known to have an enlarged anterior fontanelle. The aim of this study was to assess whether fetuses diagnosed with Down syndrome in the second trimester have larger anterior fontanelles in comparison with normal euploid fetuses. METHODS: The study population included 13 fetuses with trisomy 21 and 26 normal euploid fetuses analyzed between the 19(th) and the 23(rd) weeks of gestation. The anterior fontanelle was assessed by three-dimensional ultrasound, with the midsagittal plane of the fetal head being the reference view for acquisition of the volume. Anteroposterior and laterolateral diameters, perimeter and area of the fontanelle were then measured offline; the diameters were normalized for biparietal diameter and the perimeter and area were normalized for head circumference. Non-parametric statistical analysis was used to compare the mean values of all variables in the two groups of fetuses. Intra- and interobserver variability were also assessed. RESULTS: All variables except the fontanelle laterolateral diameter were significantly greater in Down syndrome fetuses than in controls, with anterior fontanelle perimeter/head circumference and fontanelle area/head circumference ratios showing the highest sensitivity for the detection of Down syndrome. Using a cut-off of 2.1 for the fontanelle area/head circumference ratio, the sensitivity and specificity for the detection of Down syndrome were 77% and 96%, respectively. CONCLUSION: During the mid-trimester the dimensions of the anterior fontanelle are significantly increased in fetuses with Down syndrome in comparison with normal euploid fetuses. This finding may be of help in the detection of trisomy 21 at the time of the anomaly scan.
Asunto(s)
Suturas Craneales/anomalías , Síndrome de Down/diagnóstico por imagen , Frente/diagnóstico por imagen , Suturas Craneales/ultraestructura , Síndrome de Down/embriología , Femenino , Frente/anomalías , Humanos , Recién Nacido , Masculino , Valor Predictivo de las Pruebas , Embarazo , Resultado del Embarazo , Segundo Trimestre del Embarazo , Ultrasonografía PrenatalRESUMEN
Current knowledge of suture biology has been ascertained as a result of morphological studies of normal cranial sutures (and rarely those undergoing craniosynostosis). These were initially undertaken often using histological investigations, or more recently using CT scans, as investigative tools, but have often used animal models. However, recent technological advances have provided the potential to refine our understanding of the ultrastructure by the use of new advanced scanning technology, which offers the possibility of more detailed resolution. Our aim was to undertake detailed scans of normal, fusing and fused sutures from patients with craniosynosotosis affecting different sutures, to study the detailed structure at different stages of the fusion process using a modern micro-CT scanner and a microanalytical scanning electron microscope. We wished to include in our study all the human sutures because previous studies have mostly been undertaken using the sagittal suture. Ten sutures from seven patients have revealed a complex ultra-structural arrangement. The different patterns of bone ridging seen on the ectocranial and endocranial surfaces of the fused sagittal suture were not repeated on closer inspection of either fused coronal or lambdoid sutures. Elemental analysis confirmed that the amount of calcium increased and the amount of carbon decreased as sampled areas moved away from the suture margin. We conclude that scanning allowed detailed assessment and revealed the complex arrangement of the structure of the human cranial sutures and those undergoing the process of craniosynostosis, with some differences in final structure depending on the affected suture.
Asunto(s)
Suturas Craneales/ultraestructura , Craneosinostosis/ultraestructura , Preescolar , Suturas Craneales/anatomía & histología , Suturas Craneales/diagnóstico por imagen , Craneosinostosis/diagnóstico por imagen , Femenino , Análisis de Elementos Finitos , Humanos , Lactante , Masculino , Microscopía Electrónica de Rastreo , Tomografía Computarizada por Rayos X/métodosRESUMEN
BACKGROUND: Histology remains the standard form to analyze cranial suture in murine models, but this technique provides only limited "snapshots" of the entire suture and requires animal euthanasia with tissue destruction. Because of the bone complex microarchitecture, better methods are required to study the behavior of the cranial suture and its surrounding environment. The authors compared microcomputed tomography and histology as techniques to evaluate murine cranial sutures. METHODS: A total of 360 microcomputed tomography images and 160 to 170 histologic sections were processed from a mouse at postnatal days 22 and 45, respectively. After euthanasia, the posterior frontal and sagittal sutures were imaged with a microcomputed tomography system and subsequently processed for histologic analysis. Quantitative analysis of two-dimensional images was performed to determine the percentage of bone in a 1-mm sample. RESULTS: Quantitative analysis of the percentage of bone within the sutures showed identical patterns by microcomputed tomography and histology techniques. Both methods demonstrated the posterior frontal suture to have heavier fusion patterns in the anterior and endocranial portions, with variable skip areas of complete patency on the endocranial surface, ectocranial surface, or both at day 45. CONCLUSIONS: Cranial suture fusion in the murine model is not an "all-or-none" phenomenon. The posterior frontal suture, previously thought to be completely fused on day 45 by histological analysis, showed variable fusion along the length of the suture by both methods. Quantitative assessment of the percentage of bone within the posterior frontal and sagittal sutures and morphologic assessment of these sutures demonstrated similar findings by both methods. Whereas thorough histologic evaluation of an entire suture would be extremely labor intensive and impractical, these findings help to validate microcomputed tomography as a rapid and reliable method of examining the entire suture in murine models.
Asunto(s)
Cefalometría/métodos , Suturas Craneales/ultraestructura , Ratones/anatomía & histología , Microscopía , Tomografía Computarizada por Rayos X/métodos , Animales , Suturas Craneales/diagnóstico por imagen , Suturas Craneales/crecimiento & desarrollo , Procesamiento de Imagen Asistido por Computador , Masculino , Ratones/crecimiento & desarrollo , Adhesión en Parafina , Coloración y Etiquetado , Tomografía Computarizada por Rayos X/veterinariaRESUMEN
Craniosynostosis represents a heterogeneous cluster of congenital disorders and manifests as premature ossification of one or more cranial sutures. Cranial sutures serve to enable calvarial growth and function as joints between skull bones. The mechanical properties of synostosed cranial sutures are of vital importance to their function and yet are poorly understood. The present study was designed to characterize the nanostructural and nanomechanical properties of synostosed postnatal sagittal and metopic sutures. Synostosed postnatal sagittal sutures (n = 5) and metopic sutures (n = 5) were obtained from craniosynostosis patients (aged 9.1 +/- 2.8 months). The synostosed sutural samples were prepared for imaging and indentation on both the endocranial and ectocranial surfaces with the cantilever probe of an atomic force microscopy. Analysis of the nanotopographic images indicated robust variations in sutural surface characteristics with localized peaks and valleys. In 5 x 5 mum scan sizes, the surface roughness of the synostosed metopic suture was significantly greater (223.6 +/- 93.3 nm) than the synostosed sagittal suture (142.9 +/- 80.3 nm) (P < 0.01). The Young's modulus of the synostosed sagittal suture at 0.7 +/- 0.2 MPa was significantly higher than the synostosed metopic suture at 0.5 +/- 0.1 MPa (P < 0.01). These data suggest that various synostosed cranial sutures may have different structural and mechanical characteristics.
Asunto(s)
Suturas Craneales/ultraestructura , Craneosinostosis/patología , Nanoestructuras , Nanotecnología , Fenómenos Biomecánicos , Suturas Craneales/fisiopatología , Craneosinostosis/fisiopatología , Elasticidad , Hueso Frontal/fisiopatología , Hueso Frontal/ultraestructura , Humanos , Lactante , Microscopía de Fuerza Atómica , Hueso Parietal/fisiopatología , Hueso Parietal/ultraestructura , Estrés MecánicoRESUMEN
OBJECTIVE: The aim of this experiment is to know whether active orthopedic appliance is rationale to do presurgical treatment and seek some regularities. METHODS: Laser scanning confocal microscopy and triple fluorochrome labeling in bone were used to observe and study detailed changes occurred in compressed suture systematically. RESULTS: Widened suture and broken fibres between the adjoining bones was observed at the 3rd week postoperatively. At the 3rd week hyperplasia of organic tissue components was also seen. At the 6 and 9th week there were no apparent differences among all groups no matter whether in control group or experimental groups. CONCLUSION: Retarding injury of compressed suture and surrounding tissue hyperplasia exists. With growth and development wounded suture will gradually assimilate to control group structurally and functionally.
Asunto(s)
Suturas Craneales/ultraestructura , Maxilar/ultraestructura , Animales , Suturas Craneales/patología , Femenino , Hiperplasia , Rayos Láser , Maxilar/patología , Microscopía Confocal , ConejosRESUMEN
The purpose of this study is to test predicted form-function relationships between cranial suture complexity and masticatory muscle mass and biomechanics in a mouse model. Specifically, to test the hypothesis that increased masticatory muscle mass increases sagittal suture complexity, we measured the fractal dimension (FD), temporalis mass, and temporalis bite force in myostatin-deficient (GDF8(-/-)) mice and wild-type CD-1 mice (all male, 6 months old). Myostatin is a negative regulator of muscle mass, and myostatin-deficient mice show a marked increase in muscle mass compared to normal mice. We predicted that increased sagittal suture complexity would decrease suture stiffness. The data presented here demonstrate that increased suture complexity (measured as FD) was observed in a hypermuscular mouse model (GDF8(-/-)) with significantly increased temporalis muscle mass and bite forces. Hypermuscular mice were also found to possess suture connective tissue that was less stiff (i.e., underwent more displacement before failure occurred) when loaded in tension. By decreasing stiffness, suture complexity apparently helps to dissipate mechanical loads within the cranium that are related to chewing. These results suggest that cranial suture connective tissue locally adapts to functional demands of the biomechanical suture environment. As such, cranial sutures provide a novel model for studies in connective tissue mechanotransduction.
Asunto(s)
Suturas Craneales/anatomía & histología , Suturas Craneales/fisiología , Músculo Temporal/anatomía & histología , Músculo Temporal/fisiología , Animales , Fuerza de la Mordida , Tejido Conectivo/fisiología , Suturas Craneales/ultraestructura , Elasticidad , Fractales , Masculino , Masticación/fisiología , Ratones , Ratones Endogámicos , Ratones Noqueados , Microscopía Electrónica de Rastreo , Miostatina , Tamaño de los Órganos , Osteogénesis , Estrés Mecánico , Resistencia a la Tracción , Factor de Crecimiento Transformador beta/deficienciaRESUMEN
We previously demonstrated that tensile stress (TS)-induced osteoblast differentiation eventually led to osteogenesis in an organ culture of mouse calvarial sutures. In the present study, we employed RNA-fingerprinting using an arbitrarily primed polymerase chain reaction (RAP-PCR) to identify alpha-adaptin C, a component of the endocytosis machinery AP2, as a TS-inducible gene. Protein production, as well as the gene expression of alpha-adaptin C, was induced by TS as early as 3 h following the initiation of loading. In situ hybridization and immunohistochemical analysis revealed that the induction of alpha-adaptin C mostly occurred in fibroblastic cells in the sutures, suggesting that it precedes TS-induced osteoblast differentiation. Consistent with this result, TS significantly increased the number of coated pits (CPs) and coated vesicles (CVs) in the undifferentiated fibroblastic cells but not in the osteoblastic cells around calvarial bones. Further, TS-induced osteoblast differentiation was suppressed when endocytosis was inhibited by potassium depletion. These results, taken together, suggest that TS accelerates osteoblast differentiation and osteogenesis, possibly through the induction of the alpha-adaptin C expression and consequent activation of receptor-mediated endocytosis.
Asunto(s)
Complejo 2 de Proteína Adaptadora/biosíntesis , Subunidades alfa de Complejo de Proteína Adaptadora/biosíntesis , Osteoblastos/citología , Osteogénesis , Complejo 2 de Proteína Adaptadora/análisis , Complejo 2 de Proteína Adaptadora/genética , Subunidades alfa de Complejo de Proteína Adaptadora/análisis , Subunidades alfa de Complejo de Proteína Adaptadora/genética , Animales , Diferenciación Celular , Invaginaciones Cubiertas de la Membrana Celular/ultraestructura , Vesículas Cubiertas/ultraestructura , Suturas Craneales/anatomía & histología , Suturas Craneales/metabolismo , Suturas Craneales/ultraestructura , Endocitosis , Regulación de la Expresión Génica , Ratones , Técnicas de Cultivo de Órganos , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Cráneo/citología , Estrés MecánicoRESUMEN
OBJECTIVE: Our objective was to assess the ability of the microcomputed tomography scanner to correctly image normal and synostosed cranial sutures at the ultrastructural level. DESIGN AND METHODS: Two specimens of coronal sutures were collected from operative specimens. After appropriate preparation, histological sections were obtained and stained with toluene blue for evaluation. Representative histological sections were compared to microcomputed tomography slices. RESULTS AND CONCLUSIONS: With microcomputed tomography, we successfully imaged one normal and one synostosed human coronal suture and performed a quantitative analysis of these specimens. Microcomputed tomography scanning was found to be a highly accurate imaging device for the evaluation of cranial suture development. Microcomputed tomography offers three-dimensional imaging at the microscopic level and allows for rapid quantitative analysis of bone architecture, including several measurements unavailable through histologic analysis. We believe that microcomputed tomography can play an important role in imaging and in the quantitative analysis of the stereology of bone microarchitecture. Among its advantages, microcomputed tomography is able to image many more slices than are obtainable through histology, and the method is not prone to human error. Microcomputed tomography slices are generated without destruction of the specimen and without loss or corruption of reproducible data. Structure-oriented slices from microcomputed tomography together with cellular-oriented sections from histology are complementary in the overall quantitative analysis of cranial sutures.
Asunto(s)
Suturas Craneales/ultraestructura , Craneosinostosis/patología , Microrradiografía/métodos , Tomografía Computarizada por Rayos X/métodos , Preescolar , Suturas Craneales/diagnóstico por imagen , Craneosinostosis/diagnóstico por imagen , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Lactante , Microrradiografía/instrumentación , Tomografía Computarizada por Rayos X/instrumentaciónRESUMEN
The growth and morphology of craniofacial sutures are thought to reflect their functional environment. However, little is known about in vivo sutural mechanics. The present study investigates the strains experienced by the internasal, nasofrontal, and anterior interfrontal sutures during masticatory activity in 4-6-month-old miniature swine (Sus scrofa). Measurements of the bony/fibrous arrangements and growth rates of these sutures were then examined in the context of their mechanical environment. Large tensile strains were measured in the interfrontal suture (1,036 microepsilon +/- 400 SD), whereas the posterior internasal suture was under moderate compression (-440 microepsilon +/- 238) and the nasofrontal suture experienced large compression (-1,583 microepsilon +/- 506). Sutural interdigitation was associated with compressive strain. The collagen fibers of the internasal and interfrontal sutures were clearly arranged to resist compression and tension, respectively, whereas those of the nasofrontal suture could not be readily characterized as either compression or tension resisting. The average linear rate of growth over a 1-week period at the nasofrontal suture (133.8 micrometer, +/- 50.9 S.D) was significantly greater than that of both the internasal and interfrontal sutures (39.2 micrometer +/- 11.4 and 65. 5 micrometer +/- 14.0, respectively). Histological observations suggest that the nasofrontal suture contains chondroid tissue, which may explain the unexpected combination of high compressive loading and rapid growth in this suture.
Asunto(s)
Suturas Craneales/crecimiento & desarrollo , Disostosis Craneofacial , Masticación/fisiología , Animales , Suturas Craneales/ultraestructura , Femenino , Modelos Lineales , Masculino , Estrés Mecánico , Porcinos , Porcinos EnanosRESUMEN
This study presents comparisons of the ultrastructure of synostotic and open portions of synostotic sagittal sutures using histomorphometry, scanning electron microscopy, and microcomputed tomography. By using stereologic and histomorphometric analysis, this study proposes to demonstrate evidence of the influence of biomechanical force on the suture during the process of sagittal craniosynostosis. Finally, we propose to link the pathologic changes transforming normal suture fusion to craniosynostosis with concurrent changes in the polarity of suture fusion initiation. Seven infants (four boys and three girls) with sagittal craniosynostosis, ranging in age from 1.4 to 4.8 months (mean = 3.0 months), underwent sagittal synostectomies. The synostotic bone specimens were sectioned into three regions: an open suture, partial synostosis, and complete synostosis. Microcomputed tomographic and scanning electron microscopic scanning as well as histomorphometry was performed on all specimens to obtain detailed qualitative and quantitative information regarding the trabecular microarchitecture of the synostosed suture. Microcomputed tomographic analysis determined the bone volume fraction, trabecular thickness, trabecular separation, bone surface to bone volume ratio, and anisotropy for all specimens. Our results showed significant differences in all of these quantitative measurements when comparing the complete synostotic suture with the open portion of the synostotic sutures (p < 0.05). Microcomputed tomographic stereologic analysis showed evidence of the influence of biomechanical force on the synostotic and open portions of the synostotic sutures. Results of scanning electron microscopy show a definite qualitative difference in the trabecular pattern of the partial and complete synostotic suture when compared with the open portion of the synostotic sagittal suture. In this study, we performed both qualitative and quantitative comparisons of the ultrastructure of the complete synostotic and nonsynostotic sagittal sutures using stereologic and histomorphometric techniques. We also demonstrated evidence of the influence of biomechanical force on the synostotic sagittal suture. Finally, we established a link between the pathologic changes transforming normal suture fusion to craniosynostosis and concurrent changes in both the vector and direction of suture fusion initiation.
Asunto(s)
Suturas Craneales/patología , Craneosinostosis/patología , Fenómenos Biomecánicos , Suturas Craneales/ultraestructura , Craneosinostosis/fisiopatología , Craneosinostosis/cirugía , Femenino , Humanos , Lactante , MasculinoRESUMEN
The application of transmission electron microscopy (TEM) and atomic-force microscopy (AFM) aid the acquisition of detailed structural information on the process of hard tissue formation. The sutural mineralization of rat calvaria is taken as a model for a collagen-related mineralization system. After cryofixation or chemical fixation an anhydrous tissue preparation technique with no staining procedures is used. The atomic-force microscope and the transmission electron microscope are used for structural analysis of the mineralizing region of the sutural tissue. With the application of AFM the collagen macroperiod is shown to be well represented in the unmineralized sutural tissue. At the mineralization front the collagen fibrils are found to be thickened and to change to a characteristic stacked platelet structure. Using TEM the macroperiod is faintly visible before mineral crystallites have formed and is more prominent after the apatite crystallization has started in the fibrils. In this step a needle-like structure of the newly formed apatitic crystals is visible.
Asunto(s)
Minerales/metabolismo , Cráneo/metabolismo , Cráneo/ultraestructura , Animales , Apatitas/metabolismo , Densidad Ósea , Colágeno/metabolismo , Colágeno/ultraestructura , Suturas Craneales/metabolismo , Suturas Craneales/ultraestructura , Cristalización , Microscopía de Fuerza Atómica , Microscopía Electrónica , Ratas , Ratas Sprague-DawleyRESUMEN
The cause of craniosynostosis continues to elude researchers. Although several studies have looked at the ultrastructure of normal suture closure, no previous studies have examined the microarchitecture of the synostotic suture. Our goal was to assess the scanning electron microscope (SEM) as a viable and useful tool in examining craniosynostosis. Our hypothesis is that the SEM is a powerful analytical tool that can evaluate nonsynostotic, partial synostotic, and complete synostotic cranial sutures. We analyzed the cranial suture of 3 human infants with nonsyndromic sagittal craniosynostosis. The specimens were separated into three groups, which included regions of partial and complete synostosis and a region of open suture. Histological examination provided cellular and tissue data about craniosynostosis, whereas the SEM provided detailed information regarding the trabecular microarchitecture of the synostosed suture. The SEM produced quality images of complete and partially synostotic sutures and open sutures. At low magnification, the SEM characterized the general bony microarchitecture of cranial sutures in a manner different from, but complementary to, standard histological sections. At higher magnification, the SEM allowed us a look at the cellular population of craniosynostotic sutures in a way that surpasses standard light microscopy. The SEM is an excellent tool for the study of craniosynostosis and has proved invaluable in our ability to evaluate the microarchitecture and cellular population of the fusing suture. We believe we have proven our hypothesis by demonstrating the SEM to be a powerful analytical tool that can evaluate nonsynostotic, partial synostotic, and complete synostotic cranial sutures.
Asunto(s)
Suturas Craneales/ultraestructura , Craneosinostosis/patología , Microscopía Electrónica de Rastreo/métodos , Técnicas de Preparación Histocitológica , Humanos , Lactante , Microscopía Electrónica de Rastreo/instrumentaciónRESUMEN
Morphological features of the mid-palatal suture were studied in human foetuses from 4 to 9 months of intra-uterine life. The foetuses were divided into three age groups, GI (16-23 weeks), GII (24-31 weeks) and GIII (32-39 weeks). The mid-palatal suture in GI foetuses is rectilineal in form with a wide space between the palatal processes of the maxilla. The suture has a sinuous nature in GII and GIII foetuses due to growth of the bone processes crossing the mid-line. A wide zone of cellular proliferation observed in GI narrows in GII and GIII foetuses. The imbricating nature of the suture in GII and GIII is caused by bone growth adjacent to the mid-palatal suture. Sharpey's fibres, emerging from the bone processes, run to the median region of the mid-palatal suture and are observed from GI foetuses onwards. The collagen fibres of the mid-palatal suture are orientated transversely under the oral epithelium and exhibit a regular meshwork with a predominance of sagittal fibres in the median region of the suture. These fibres are orientated transversely and obliquely at the junction with the nasal septum.
Asunto(s)
Suturas Craneales/anatomía & histología , Suturas Craneales/embriología , Hueso Paladar/anatomía & histología , Hueso Paladar/embriología , Colágeno/ultraestructura , Suturas Craneales/ultraestructura , Femenino , Feto/anatomía & histología , Edad Gestacional , Humanos , Masculino , Microscopía Electrónica de Rastreo , Hueso Paladar/ultraestructuraRESUMEN
Syndesmotic sutures of the skull are formed by dense connective tissue and called "open"; they are "closed" by formation of a synostosis between adjacent bones. Open sutures are considered as areas of growth and as hinges. The importance of open sutures during the period of skull growth is reflected by pathological situations in which premature closure of the sutures occurs. As alterations of the FGF receptor have been reported in genetical disorders accompanied by premature suture closure (Bellus et al. 1996), the role of fibroblasts and connective tissue in the development of the sagittal suture of mice has been investigated by light and electron microscopy. Morphological changes of the sagittal suture at the following stages are reported: at embryonic day 18, days 1, 5, 9, 14, 20, 26, 28 after birth and in adult mice. Two skulls per stage were investigated. Early osteogenesis appeared in a thin plate, followed by a second plate underneath the first one. Both were separated by blood vessels. In general, vascularization preceded desmoid mineralization; the space around blood vessels was occupied by non-bone-forming cells leaving cavities for the presumptive bone marrow. Mineralization of the collagen-rich osteoid at the mineralizing rim of the bone plates was accompanied by apoptoses and cell disintegration. Newly formed bone was immediately covered by osteoblasts forming a sheet of bone-lining cells. At day 9, the double-layered bone plates of both sides reached the median area of the skull but were separated by non-mineralizing, collagen-rich connective tissue. From day 14 onwards, the bone plates thickened. Bone apposition, recognizable by the formation of collagen-rich osteoid and proceeding from day 14 pp onwards, occurred mainly at the outer and inner surfaces of the calvariae, but neither at bone marrow surfaces nor at the medial edges of the parietal bones. These opposite bone faces showed fewer osteoblasts and bone-lining cells, but an increased number of fibroblasts. Tendon-like collagen bundles connected both bone plates of the open suture of day 26 pp as well as in the adult mice, whereby synostotically closed areas alternated. Formation of an open, syndesmotic suture can, therefore, be described as a transition of bone-forming tissue into a bone-tendon junction. The results indicate the importance of the replacement of osteoblasts by fibroblasts at the sutural front of the bone plates in order to prevent a premature suture closure.
Asunto(s)
Suturas Craneales/crecimiento & desarrollo , Suturas Craneales/ultraestructura , Osteogénesis/fisiología , Hueso Parietal/crecimiento & desarrollo , Hueso Parietal/ultraestructura , Animales , Animales Recién Nacidos , Femenino , Ratones , EmbarazoRESUMEN
Many bones in mammalian skulls are linked together by cranial sutures, connective tissue joints that are morphologically variable and show different levels of interdigitation among and within species. The goal of this investigation was to determine whether sections of skull with cranial sutures have different mechanical properties than adjacent sections without sutures, and if these properties are enhanced with increased interdigitation. To test these hypotheses, bending strength and impact energy absorption were measured for samples of goat (Capra hircus) cranial bone without sutures and with sutures of different degrees of interdigitation. Bending strength was measured under both dynamic (9.7 mm displacement s-1) and relatively static (0.8 mm s-1) conditions, and at either speed, increased sutural interdigitation provided increased strength during three-point bending. However, except for very highly interdigitated sutures loaded slowly, sutures were not as strong in bending as bone. In contrast, sutures absorbed from 16% to 100% more energy per unit volume during impact loading than did bone. This five-fold increase in energy absorption by the sutures was significantly correlated with increased sutural interdigitation.
Asunto(s)
Suturas Craneales/fisiología , Cráneo/fisiología , Animales , Fenómenos Biomecánicos , Suturas Craneales/ultraestructura , Femenino , Cabras , Técnicas In VitroRESUMEN
Morphogenesis and maturation of the sagittal suture in newborn C57B1/6J strain mice were studied using light and electron microscopy. Morphodifferentiation of the murine parietal bones progresses radially with the interposed sagittal suture, assuming a greater level of maturity at birth at a midpoint along its length. The presumptive suture develops in a sulcus, deeper posteriorly, more shallow anteriorly. Cells at the osteogenic front (OF) are distinguished from the surrounding fibrocytic cells by a number of distinctive characteristics: 1. increased cytoplasmic density; 2. extensive endoplasmic reticulum; 3. dispersed nuclear chromatin aggregates; 4. extensive surface projections; 5. close approximation. Mineralization of the developing parietal bone occurs extracellularly with the initial deposits of apatite crystals exhibiting no oriented relationship to either cellular or extracellular fibrillar elements. The majority of collagen fibers lie superior and inferior to the presumptive suture, oriented anteroposteriorly with their long axes parallel to the ectocranial surface. Other fiber bundles more intimately associated with the developing suture display a more random orientation.