RESUMO
OBJECTIVE: To explore the impact of oligohydramnios on fetal movement and hip development, given its association with developmental dysplasia of the hip (DDH) but unclear mechanisms. METHODS: Chick embryos were divided into four groups based on the severity of oligohydramnios induced by amniotic fluid aspiration (control, 0.2 mL, 0.4 mL, 0.6 mL). Fetal movement was assessed by detection of movement and quantification of residual amniotic fluid volume. Hip joint development was assessed by gross anatomic analysis, micro-computed tomography (micro-CT) for cartilage assessment, and histologic observation at multiple time points. In addition, a subset of embryos from the 0.4 mL aspirated group underwent saline reinfusion and subsequent evaluation. RESULTS: Increasing volumes of aspirated amniotic fluid resulted in worsening of fetal movement restrictions (e.g., 0.4 mL aspirated and control group at E10: frequency difference -7.765 [95% CI: -9.125, -6.404]; amplitude difference -0.343 [95% CI: -0.588, -0.097]). The 0.4 mL aspirated group had significantly smaller hip measurements compared to controls, with reduced acetabular length (-0.418 [95% CI: -0.575, -0.261]) and width (-0.304 [95% CI: -0.491, -0.117]) at day E14.5. Histological analysis revealed a smaller femoral head (1.084 ± 0.264 cm) and shallower acetabulum (0.380 ± 0.106 cm) in the 0.4 mL group. Micro-CT showed cartilage matrix degeneration (13.6% [95% CI: 0.6%, 26.7%], P = 0.043 on E14.5). Saline reinfusion resulted in significant improvements in the femoral head to greater trochanter (0.578 [95% CI: 0.323, 0.833], P = 0.001). CONCLUSIONS: Oligohydramnios can cause DDH by restricting fetal movement and disrupting hip morphogenesis in a time-dependent manner. Timely reversal of oligohydramnios during the fetal period may prevent DDH.
Assuntos
Displasia do Desenvolvimento do Quadril , Modelos Animais de Doenças , Oligo-Hidrâmnio , Microtomografia por Raio-X , Animais , Embrião de Galinha , Oligo-Hidrâmnio/diagnóstico por imagem , Displasia do Desenvolvimento do Quadril/diagnóstico por imagem , Movimento Fetal , Articulação do Quadril/diagnóstico por imagem , Articulação do Quadril/patologia , Articulação do Quadril/embriologia , Feminino , Cartilagem Articular/diagnóstico por imagem , Cartilagem Articular/patologia , Cartilagem Articular/embriologia , Líquido Amniótico , GravidezRESUMO
Although the knee joint (KNJ) and temporomandibular joint (TMJ) all belong to the synovial joint, there are many differences in developmental origin, joint structure and articular cartilage type. Studies of joint development in embryos have been performed, mainly using poultry and rodents. However, KNJ and TMJ in poultry and rodents differ from those in humans in several ways. Very little work has been done on the embryonic development of KNJ and TMJ in large mammals. Several studies have shown that pigs are ideal animals for embryonic development research. Embryonic day 30 (E30), E35, E45, E55, E75, E90, Postnatal day 0 (P0) and Postnatal day 30 (P30) embryos/fetuses from the pigs were used for this study. The results showed that KNJ develops earlier than TMJ. Only one mesenchymal condensate of KNJ is formed on E30, while two mesenchymal condensates of TMJ are present on E35. All structures of KNJ and TMJ were formed on E45. The growth plate of KNJ begins to develop on E45 and becomes more pronounced from E55 to P30. From E75 to E90, more and more vascular-rich cartilage canals form in the cartilage regions of both joints. The cartilaginous canal of the TMJ divides the condyle into sections along the longitudinal axis of the condyle. This arrangement of cartilaginous canal was not found in the KNJ. The chondrification of KNJ precedes that of TMJ. Ossification of the knee condyle occurs gradually from the middle to the periphery, while that of the TMJ occurs gradually from the base of the mandibular condyle. In the KNJ, the ossification of the articular condyle is evident from P0 to P30, and the growth plate is completely formed on P30. In the TMJ, the cartilage layer of condyle becomes thinner from P0 to P30. There is no growth plate formation in TMJ during its entire development. There is no growth plate formation in the TMJ throughout its development. The condyle may be the developmental center of the TMJ. The chondrocytes and hypertrophic chondrocytes of the growth plate are densely arranged. The condylar chondrocytes of TMJ are scattered, while the hypertrophic chondrocytes are arranged. Embryonic development of KNJ and TMJ in pigs is an important bridge for translating the results of rodent studies to medical applications.
Assuntos
Articulação do Joelho , Articulação Temporomandibular , Animais , Suínos/embriologia , Articulação Temporomandibular/embriologia , Articulação Temporomandibular/crescimento & desenvolvimento , Articulação do Joelho/embriologia , Articulação do Joelho/crescimento & desenvolvimento , Cartilagem Articular/embriologia , Cartilagem Articular/crescimento & desenvolvimento , Feminino , Desenvolvimento Embrionário/fisiologia , Embrião de MamíferosRESUMO
Previous studies on mouse embryo limbs have established that interzone mesenchymal progenitor cells emerging at each prescribed joint site give rise to joint tissues over fetal time. These incipient tissues undergo structural maturation and morphogenesis postnatally, but underlying mechanisms of regulation remain unknown. Hox11 genes dictate overall zeugopod musculoskeletal patterning and skeletal element identities during development. Here we asked where these master regulators are expressed in developing limb joints and whether they are maintained during postnatal zeugopod joint morphogenesis. We found that Hoxa11 was predominantly expressed and restricted to incipient wrist and ankle joints in E13.5 mouse embryos, and became apparent in medial and central regions of knees by E14.5, though remaining continuously dormant in elbow joints. Closer examination revealed that Hoxa11 initially characterized interzone and neighboring cells and was then restricted to nascent articular cartilage, intra joint ligaments and structures such as meniscal horns over prenatal time. Postnatally, articular cartilage progresses from a nondescript cell-rich, matrix-poor tissue to a highly structured, thick, zonal and mechanically competent tissue with chondrocyte columns over time, most evident at sites such as the tibial plateau. Indeed, Hox11 expression (primarily Hoxa11) was intimately coupled to such morphogenetic processes and, in particular, to the topographical rearrangement of chondrocytes into columns within the intermediate and deep zones of tibial plateau that normally endures maximal mechanical loads. Revealingly, these expression patterns were maintained even at 6 months of age. In sum, our data indicate that Hox11 genes remain engaged well beyond embryonic synovial joint patterning and are specifically tied to postnatal articular cartilage morphogenesis into a zonal and resilient tissue. The data demonstrate that Hox11 genes characterize adult, terminally differentiated, articular chondrocytes and maintain region-specificity established in the embryo.
Assuntos
Cartilagem Articular/embriologia , Condrogênese/genética , Genes Homeobox , Membrana Sinovial/embriologia , Animais , Condrogênese/fisiologia , Extremidades/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Genes Reporter , Proteínas de Fluorescência Verde/genética , CamundongosRESUMO
Adult articular cartilage shows limited tissue turnover, and therefore development of the proper structure of articular cartilage is crucial for life-long joint function. However, the mechanism by which the articular cartilage structure is developmentally regulated is poorly understood. In this study, we show evidence that activation of extracellular signal-regulated kinases (Erk1/2) in articular chondrocyte progenitors during developmental stages control articular cartilage thickness. We found that overexpression of Lin28a, an RNA-binding protein that regulates organismal growth and metabolism, in articular chondrocyte progenitor cells upregulated Erk signaling and increased articular cartilage thickness. Overexpression of a constitutively active Kras mimicked Lin28a overexpression, and inhibition of Erk signaling during embryonic stages normalized the cartilage phenotype of both Kras- and Lin28a-overexpressing mice. These results suggest that articular cartilage thickness is mainly determined during the process of embryonic synovial joint development, which is positively regulated by Erk signaling.
Assuntos
Cartilagem Articular/embriologia , Condrogênese/genética , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas de Ligação a RNA/genética , Animais , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Condrócitos/fisiologia , Embrião de Mamíferos , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Sistema de Sinalização das MAP Quinases/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Gravidez , Transdução de Sinais/genéticaRESUMO
Dynamic mechanical loading of synovial joints is necessary for normal joint development, as evidenced in certain clinical conditions, congenital disorders and animal models where dynamic muscle contractions are reduced or absent. Although the importance of mechanical forces on joint development is unequivocal, little is known about the molecular mechanisms involved. Here, using chick and mouse embryos, we observed that molecular changes in expression of multiple genes analyzed in the absence of mechanical stimulation are consistent across species. Our results suggest that abnormal joint development in immobilized embryos involves inappropriate regulation of Wnt and BMP signaling during definition of the emerging joint territories, i.e. reduced ß-catenin activation and concomitant upregulation of pSMAD1/5/8 signaling. Moreover, dynamic mechanical loading of the developing knee joint activates Smurf1 expression; our data suggest that Smurf1 insulates the joint region from pSMAD1/5/8 signaling and is essential for maintenance of joint progenitor cell fate.
Assuntos
Padronização Corporal , Proteínas Morfogenéticas Ósseas/metabolismo , Articulações/embriologia , Articulações/metabolismo , Movimento/fisiologia , Animais , Padronização Corporal/genética , Proteínas Morfogenéticas Ósseas/genética , Cartilagem Articular/embriologia , Cartilagem Articular/metabolismo , Diferenciação Celular/genética , Embrião de Galinha , Embrião de Mamíferos , Regulação da Expressão Gênica no Desenvolvimento , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Transdução de Sinais/genética , beta Catenina/genética , beta Catenina/metabolismoRESUMO
During appendicular skeletal development, the bi-potential cartilage anlagen gives rise to transient cartilage, which is eventually replaced by bone, and to articular cartilage that caps the ends of individual skeletal elements. While the molecular mechanism that regulates transient cartilage differentiation is relatively well understood, the mechanism of articular cartilage differentiation has only begun to be unraveled. Furthermore, the molecules that coordinate the articular and transient cartilage differentiation processes are poorly understood. Here, we have characterized in chick the regulatory roles of two transcription factors, NFIA and GATA3, in articular cartilage differentiation, maintenance and the coordinated differentiation of articular and transient cartilage. Both NFIA and GATA3 block hypertrophic differentiation. Our results suggest that NFIA is not sufficient but necessary for articular cartilage differentiation. Ectopic activation of GATA3 promotes articular cartilage differentiation, whereas inhibition of GATA3 activity promotes transient cartilage differentiation at the expense of articular cartilage. We propose a novel transcriptional circuitry involved in embryonic articular cartilage differentiation, maintenance and its crosstalk with the transient cartilage differentiation program.
Assuntos
Proteínas Aviárias/metabolismo , Cartilagem Articular/embriologia , Cartilagem Articular/metabolismo , Fator de Transcrição GATA3/metabolismo , Fatores de Transcrição NFI/metabolismo , Animais , Animais Geneticamente Modificados , Proteínas Aviárias/deficiência , Proteínas Aviárias/genética , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Embrião de Galinha , Condrócitos/citologia , Condrócitos/metabolismo , Feminino , Fator de Transcrição GATA3/genética , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Camundongos Knockout , Modelos Biológicos , Fatores de Transcrição NFI/deficiência , Fatores de Transcrição NFI/genética , Gravidez , RNA Interferente Pequeno/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
Within each synovial joint, the articular cartilage is uniquely adapted to bear dynamic compressive loads and shear forces throughout the joint's range of motion. Injury and age-related degeneration of the articular cartilage often lead to significant pain and disability, as the intrinsic repair capability of the tissue is extremely limited. Current surgical and biological treatment options have been unable to restore cartilage de novo. Before successful clinical cartilage restoration strategies can be developed, a better understanding of how the cartilage forms during normal development is essential. This review focuses on recent progress made towards addressing key questions about articular cartilage morphogenesis, including the origin of synovial joint progenitor cells, postnatal development and growth of the tissue. These advances have provided novel insight into fundamental questions about the developmental biology of articular cartilage, as well as potential cell sources that may participate in joint response to injury.
Assuntos
Envelhecimento/fisiologia , Cartilagem Articular/embriologia , Desenvolvimento Embrionário , Articulações/embriologia , Animais , Humanos , Morfogênese , Células-Tronco/citologiaRESUMO
The temporomandibular joint (TMJ) is an intricate structure composed of the mandibular condyle, articular disc, and glenoid fossa in the temporal bone. Apical condylar cartilage is classified as a secondary cartilage, is fibrocartilaginous in nature, and is structurally distinct from growth plate and articular cartilage in long bones. Condylar cartilage is organized in distinct cellular layers that include a superficial layer that produces lubricants, a polymorphic/progenitor layer that contains stem/progenitor cells, and underlying layers of flattened and hypertrophic chondrocytes. Uniquely, progenitor cells reside near the articular surface, proliferate, undergo chondrogenesis, and mature into hypertrophic chondrocytes. During the past decades, there has been a growing interest in the molecular mechanisms by which the TMJ develops and acquires its unique structural and functional features. Indian hedgehog (Ihh), which regulates skeletal development including synovial joint formation, also plays pivotal roles in TMJ development and postnatal maintenance. This review provides a description of the many important recent advances in Hedgehog (Hh) signaling in TMJ biology. These include studies that used conventional approaches and those that analyzed the phenotype of tissue-specific mouse mutants lacking Ihh or associated molecules. The recent advances in understanding the molecular mechanism regulating TMJ development are impressive and these findings will have major implications for future translational medicine tools to repair and regenerate TMJ congenital anomalies and acquired diseases, such as degenerative damage in TMJ osteoarthritic conditions.
Assuntos
Cartilagem Articular/embriologia , Condrogênese , Proteínas Hedgehog/metabolismo , Osteoartrite/embriologia , Transdução de Sinais , Disco da Articulação Temporomandibular/embriologia , Animais , Cartilagem Articular/patologia , Diferenciação Celular , Humanos , Côndilo Mandibular/embriologia , Côndilo Mandibular/patologia , Camundongos , Osteoartrite/patologia , Disco da Articulação Temporomandibular/patologiaRESUMO
Limb synovial joints are composed of distinct tissues, but it is unclear which progenitors produce those tissues and how articular cartilage acquires its functional postnatal organization characterized by chondrocyte columns, zone-specific cell volumes and anisotropic matrix. Using novel Gdf5CreERT2 (Gdf5-CE), Prg4-CE and Dkk3-CE mice mated to R26-Confetti or single-color reporters, we found that knee joint progenitors produced small non-migratory progenies and distinct local tissues over prenatal and postnatal time. Stereological imaging and quantification indicated that the columns present in juvenile-adult tibial articular cartilage consisted of non-daughter, partially overlapping lineage cells, likely reflecting cell rearrangement and stacking. Zone-specific increases in cell volume were major drivers of tissue thickening, while cell proliferation or death played minor roles. Second harmonic generation with 2-photon microscopy showed that the collagen matrix went from being isotropic and scattered at young stages to being anisotropic and aligned along the cell stacks in adults. Progenitor tracing at prenatal or juvenile stages showed that joint injury provoked a massive and rapid increase in synovial Prg4+ and CD44+/P75+ cells some of which filling the injury site, while neighboring chondrocytes appeared unresponsive. Our data indicate that local cell populations produce distinct joint tissues and that articular cartilage growth and zonal organization are mainly brought about by cell volume expansion and topographical cell rearrangement. Synovial Prg4+ lineage progenitors are exquisitely responsive to acute injury and may represent pioneers in joint tissue repair.
Assuntos
Cartilagem Articular , Tamanho Celular , Condrogênese/fisiologia , Traumatismos do Joelho/metabolismo , Articulação do Joelho/crescimento & desenvolvimento , Células-Tronco Mesenquimais/metabolismo , Animais , Cartilagem Articular/citologia , Cartilagem Articular/embriologia , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/lesões , Diferenciação Celular/fisiologia , Linhagem da Célula , Proliferação de Células , Condrócitos/citologia , Colágeno/metabolismo , Fator 5 de Diferenciação de Crescimento/metabolismo , Articulação do Joelho/citologia , Camundongos , Camundongos Transgênicos , Membrana Sinovial/citologiaRESUMO
The articular cartilage, which lines the joints of the limb skeleton, is distinct from the adjoining transient cartilage, and yet, it differentiates as a unique population within a contiguous cartilage element. Current literature suggests that articular cartilage and transient cartilage originate from different cell populations. Using a combination of lineage tracing and pulse-chase of actively proliferating chondrocytes, we here demonstrate that, similar to transient cartilage, embryonic articular cartilage cells also originate from the proliferating chondrocytes situated near the distal ends of skeletal anlagen. We show that nascent cartilage cells are capable of differentiating as articular or transient cartilage, depending on exposure to Wnt or BMP signaling, respectively. The spatial organization of the articular cartilage results from a band of Nog-expressing cells, which insulates these proliferating chondrocytes from BMP signaling and allows them to differentiate as articular cartilage under the influence of Wnt signaling emanating from the interzone. Through experiments conducted in both chick and mouse embryos we have developed a model explaining simultaneous growth and differentiation of transient and articular cartilage in juxtaposed domains.
Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Proteínas de Transporte/metabolismo , Cartilagem Articular/embriologia , Linhagem da Célula/fisiologia , Transdução de Sinais/fisiologia , Azul Alciano , Animais , Cartilagem Articular/citologia , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Embrião de Galinha , Condrócitos/metabolismo , Condrócitos/fisiologia , Imuno-Histoquímica , Hibridização In Situ , Camundongos , Camundongos Transgênicos , Proteínas Wnt/metabolismoRESUMO
Joints connect the skeletal components and enable movement. The appearance and development of articulations is due to different genetic, biochemical, and mechanical factors. In the embryonic stage, controlled biochemical processes are critical for organized growth. We developed a computational model, which predicts the appearance, location, and development of joints in the embryonic stage. Biochemical events are modeled with reaction diffusion equations with generic molecules representing molecules that 1) determine the site where the articulation will appear, 2) promote proliferation, and matrix synthesis, and 3) define articular cartilage. Our model accounts for cell differentiation from mesenchymal cells to pre-cartilaginous cells, then cartilaginous cells, and lastly articular cartilage. These reaction-diffusion equations were solved using the finite elements method. From a mesenchymal 'bud' of a phalanx, the model predicts growth, joint cleavage, joint morphology, and articular cartilage formation. Our prediction of the gene expression during development agrees with molecular expression profiles of joint development reported in literature. Our computational model suggests that initial rudiment dimensions affect diffusion profiles result in Turing patterns that dictate sites of cleavage thereby determining the number of joints in a rudiment.
Assuntos
Desenvolvimento Ósseo/fisiologia , Cartilagem Articular/embriologia , Simulação por Computador , Articulações/embriologia , Animais , Biomarcadores/metabolismo , Osso e Ossos/embriologia , Osso e Ossos/metabolismo , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/fisiologia , Comunicação Celular/fisiologia , Diferenciação Celular , Proliferação de Células , Condrogênese/fisiologia , Biologia Computacional , Falanges dos Dedos da Mão/embriologia , Falanges dos Dedos da Mão/crescimento & desenvolvimento , Falanges dos Dedos da Mão/metabolismo , Fator 5 de Diferenciação de Crescimento/administração & dosagem , Fator 5 de Diferenciação de Crescimento/farmacocinética , Humanos , Articulações/citologia , Articulações/crescimento & desenvolvimento , Articulações/metabolismo , Modelos Teóricos , Morfogênese/fisiologiaRESUMO
As a key molecule of the extracellular matrix, laminin provides a delicate microenvironment for cell functions. Recent findings suggest that laminins expressed by cartilage-forming cells (chondrocytes, progenitor cells and stem cells) could promote chondrogenesis. However, few papers outline the effect of laminins on providing a favorable matrix microenvironment for cartilage regeneration. In this review, we delineated the expression of laminins in hyaline cartilage, fibrocartilage and cartilage-like tissue (nucleus pulposus) throughout several developmental stages. We also examined the effect of laminins on the biological activities of chondrocytes, including adhesion, migration and survival. Furthermore, we scrutinized the potential influence of various laminin isoforms on cartilage-forming cells' proliferation and chondrogenic differentiation. With this information, we hope to facilitate the understanding of the spatial and temporal interactions between cartilage-forming cells and laminin microenvironment to eventually advance cell-based cartilage engineering and regeneration.
Assuntos
Cartilagem Articular/embriologia , Cartilagem Articular/metabolismo , Laminina/metabolismo , Regeneração/fisiologia , Animais , Cartilagem Articular/citologia , Proliferação de Células , Condrócitos/metabolismo , Condrogênese , HumanosRESUMO
Epidemiological data show that osteoarthritis (OA) is significantly associated with lower birth weight, and that OA may be a type of fetal-originated adult disease. The present study aimed to investigate the prenatal food-restriction (PFR) effect on the quality of articular cartilage in female offspring to explore the underlying mechanisms of fetal-originated OA. Maternal rats were fed a restricted diet from gestational day (GD) 11 to 20 to induce intra-uterine growth retardation. Female fetuses and female adult offspring fed a post-weaning high-fat diet were killed at GD20 and postnatal week 24, respectively. Serum and knee cartilage samples from fetuses and adult female offspring were collected and examined for cholesterol metabolism and histology. Fetal serum corticosterone and insulin-like growth factor-1 (IGF-1) in the PFR group were lower than those of the control, but the serum cholesterol level was not changed. The lower expression of IGF-1 in the PFR group lasted into adulthood. The expression of extracellular matrix (ECM) genes, including type II collagen, aggrecan and cholesterol efflux genes including liver X receptor, were significantly induced, but the ATP-binding-cassette transporter A1 was unchanged. PFR could induce a reduction in ECM synthesis and impaired cholesterol efflux in female offspring, and eventually led to poor quality of articular cartilage and OA.
Assuntos
Doenças das Cartilagens/etiologia , Cartilagem Articular/patologia , Modelos Animais de Doenças , Proteínas da Matriz Extracelular/metabolismo , Desenvolvimento Fetal , Retardo do Crescimento Fetal/fisiopatologia , Osteoartrite do Joelho/etiologia , Animais , Restrição Calórica/efeitos adversos , Doenças das Cartilagens/embriologia , Doenças das Cartilagens/metabolismo , Doenças das Cartilagens/patologia , Cartilagem Articular/embriologia , Cartilagem Articular/metabolismo , Colesterol/sangue , Colesterol/metabolismo , Dieta Hiperlipídica/efeitos adversos , Dislipidemias/etiologia , Dislipidemias/fisiopatologia , Proteínas da Matriz Extracelular/genética , Feminino , Retardo do Crescimento Fetal/etiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Fator de Crescimento Insulin-Like I/análise , Fenômenos Fisiológicos da Nutrição Materna , Osteoartrite do Joelho/embriologia , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Gravidez , Distribuição Aleatória , Ratos Wistar , Organismos Livres de Patógenos Específicos , DesmameRESUMO
Clinical efforts to repair damaged articular cartilage (AC) currently face major obstacles due to limited intrinsic repair capacity of the tissue and unsuccessful biological interventions. This highlights a need for better therapeutic strategies. This review summarizes the recent advances in the field of cell-based AC repair. In both animals and humans, AC defects that penetrate into the subchondral bone marrow are mainly filled with fibrocartilaginous tissue through the differentiation of bone marrow mesenchymal stem cells (MSCs), followed by degeneration of repaired cartilage and osteoarthritis (OA). Cell therapy and tissue engineering techniques using culture-expanded chondrocytes, bone marrow MSCs, or pluripotent stem cells with chondroinductive growth factors may generate cartilaginous tissue in AC defects but do not form hyaline cartilage-based articular surface because repair cells often lose chondrogenic activity or result in chondrocyte hypertrophy. The new evidence that AC and synovium develop from the same pool of precursors with similar gene profiles and that synovium-derived chondrocytes have stable chondrogenic activity has promoted use of synovium as a new cell source for AC repair. The recent finding that NFAT1 and NFAT2 transcription factors (TFs) inhibit chondrocyte hypertrophy and maintain metabolic balance in AC is a significant advance in the field of AC repair. The use of synovial MSCs and discovery of upstream transcriptional regulators that help maintain the AC phenotype have opened new avenues to improve the outcome of AC regeneration.
Assuntos
Cartilagem Articular/fisiologia , Condrócitos/transplante , Condrogênese/fisiologia , Transplante de Células-Tronco Mesenquimais , Osteoartrite/terapia , Regeneração/fisiologia , Membrana Sinovial/fisiologia , Cartilagem Articular/embriologia , Cartilagem Articular/lesões , Terapia Baseada em Transplante de Células e Tecidos , Condrócitos/citologia , Humanos , Fatores de Transcrição NFATC/fisiologia , Membrana Sinovial/embriologia , Engenharia TecidualRESUMO
Articular cartilage has obvious and fundamental roles in joint function and body movement. Much is known about its organization, extracellular matrix, and phenotypic properties of its cells, but less is known about its developmental biology. Incipient articular cartilage in late embryos and neonates is a thin tissue with scanty matrix and small cells, while adult tissue is thick and zonal and contains large cells and abundant matrix. What remains unclear is not only how incipient articular cartilage forms, but how it then grows and matures into a functional, complex, and multifaceted structure. This review focuses on recent and exciting discoveries on the developmental biology and growth of articular cartilage, frames them within the context of classic studies, and points to lingering questions and research goals. Advances in this research area will have significant relevance to basic science, and also considerable translational value to design superior cartilage repair and regeneration strategies.
Assuntos
Cartilagem Articular/embriologia , Condrócitos , Matriz Extracelular , Animais , Cartilagem Articular/crescimento & desenvolvimento , Linhagem da Célula , Regeneração Tecidual Guiada , HumanosRESUMO
OBJECTIVE: The aim of this study was to test the possible involvement, relevance and significance of dentin matrix protein 1 (DMP1) in chondrocyte redifferentiation and OA. METHODS: To examine the function of DMP1 in vitro, bone marrow stromal cells (BMSCs) and articular chondrocytes (ACs) were isolated and differentiated in micromasses in the presence or absence of DMP1 small interfering RNA and analysed for chondrogenic phenotype. The association of DMP1 expression with OA progression was analysed time dependently in the OA menisectomy rat model and in grade-specific OA human samples. RESULTS: It was found that DMP1 was strongly related to chondrogenesis, which was evidenced by the strong expression of DMP1 in the 14.5-day mouse embryonic cartilage development stage and in femoral heads of post-natal days 0 and 4. In vitro chondrogenesis in BMSCs and ACs was accompanied by a gradual increase in DMP1 expression at both the gene and protein levels. In addition, knockdown of DMP1 expression led to decreased chondrocyte marker genes, such as COL2A1, ACAN and SOX9, and an increase in the expression of COL10A and MMP13 in ACs. Moreover, treatment with IL-1ß, a well-known catabolic culprit of proteoglycan matrix loss, significantly reduced the expression of DMP1. Furthermore, we also observed the suppression of DMP1 protein in a grade-specific manner in knee joint samples from patients with OA. In the menisectomy-induced OA model, an increase in the Mankin score was accompanied by the gradual loss of DMP1 expression. CONCLUSION: Observations from this study suggest that DMP1 may play an important role in maintaining the chondrogenic phenotype and its possible involvement in altered cartilage matrix remodelling and degradation in disease conditions like OA.
Assuntos
Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Proteínas da Matriz Extracelular/fisiologia , Osteoartrite do Joelho/metabolismo , Fosfoproteínas/fisiologia , Animais , Artrite Experimental/metabolismo , Cartilagem Articular/embriologia , Cartilagem Articular/patologia , Diferenciação Celular/fisiologia , Células Cultivadas , Condrócitos/patologia , Condrogênese/fisiologia , Progressão da Doença , Desenvolvimento Embrionário/fisiologia , Proteínas da Matriz Extracelular/deficiência , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Interleucina-1beta/farmacologia , Masculino , Camundongos , Osteoartrite do Joelho/patologia , Fosfoproteínas/deficiência , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , Ratos Endogâmicos WKYRESUMO
Several studies have focused on the cartilaginous, articular, and ligamentous development of the craniovertebral joint (CVJ), but there are no unifying criteria regarding the origin and morphogenetic timetable of the structures that make up the CVJ. In our study, serial sections of 53 human embryonic (n = 27) and fetal (n = 26) specimens from O'Rahilly stages 17-23 and 9-13 weeks, respectively, have been analyzed. Our results demonstrate that the chondrification of the pars basioccipitalis and exoccipitalis becomes observable at stage 19, and all future bones in the CVJ are in their cartilaginous form except for the future odontoid process. In addition, two chondrification centers appear for the body of the axis. From stage 21, the apical, alar, and transverse atlantal ligaments begin to acquire a ligamentous structure and the odontoid process initiates its chondrogenic phase. Stage 22 witnesses the first signs of the articular cavities of the atlanto-occipital joint, and by stage 23 all joints have cavities except for the transverse-odontoid joint, which will wait until week 9. In week 10, the ossification of the basilar part of the occipital bone begins, followed by the rest of the structures except for the odontoid process, which will start at week 13, thus completing the osteogenesis of all bones in the CVJ. The results of this study could help in establishing the anatomical basis of the normally functioning CVJ and for detecting its related pathologies, abnormalities, and malformations.
Assuntos
Articulação Atlantoaxial/embriologia , Articulação Atlantoccipital/embriologia , Cartilagem Articular/embriologia , Feto/anatomia & histologia , Forame Magno/embriologia , Ligamentos Articulares/embriologia , Humanos , Morfogênese , Osso Occipital/embriologiaRESUMO
SOX9 is a transcription factor of the SRY family that regulates sex determination, cartilage development and numerous other developmental events. In the foetal growth plate, Sox9 is highly expressed in chondrocytes of the proliferating and prehypertrophic zone but declines abruptly in the hypertrophic zone, suggesting that Sox9 downregulation in hypertrophic chondrocytes might be a necessary step to initiate cartilage-bone transition in the growth plate. In order to test this hypothesis, we generated transgenic mice misexpressing Sox9 in hypertrophic chondrocytes under the control of a BAC-Col10a1 promoter. The transgenic offspring showed an almost complete lack of bone marrow in newborns, owing to strongly retarded vascular invasion into hypertrophic cartilage and impaired cartilage resorption, resulting in delayed endochondral bone formation associated with reduced bone growth. In situ hybridization analysis revealed high levels of Sox9 misexpression in hypertrophic chondrocytes but deficiencies of Vegfa, Mmp13, RANKL and osteopontin expression in the non-resorbed hypertrophic cartilage, indicating that Sox9 misexpression in hypertrophic chondrocytes inhibits their terminal differentiation. Searching for the molecular mechanism of SOX9-induced inhibition of cartilage vascularization, we discovered that SOX9 is able to directly suppress Vegfa expression by binding to SRY sites in the Vegfa gene. Postnatally, bone marrow formation and cartilage resorption in transgenic offspring are resumed by massive invasion of capillaries through the cortical bone shaft, similar to secondary ossification. These findings imply that downregulation of Sox9 in the hypertrophic zone of the normal growth plate is essential for allowing vascular invasion, bone marrow formation and endochondral ossification.
Assuntos
Medula Óssea/embriologia , Calcificação Fisiológica/genética , Cartilagem/irrigação sanguínea , Neovascularização Fisiológica/genética , Fatores de Transcrição SOX9/fisiologia , Animais , Animais Recém-Nascidos , Medula Óssea/metabolismo , Cartilagem/metabolismo , Cartilagem Articular/embriologia , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/patologia , Cartilagem Articular/fisiologia , Diferenciação Celular/genética , Condrócitos/metabolismo , Condrócitos/patologia , Condrócitos/fisiologia , Embrião de Mamíferos , Regulação da Expressão Gênica no Desenvolvimento , Lâmina de Crescimento/embriologia , Lâmina de Crescimento/crescimento & desenvolvimento , Lâmina de Crescimento/metabolismo , Hipertrofia/genética , Camundongos , Camundongos Transgênicos , Organogênese/genética , Osteogênese/genética , Osteogênese/fisiologia , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismoRESUMO
Cartilage oligomeric matrix protein (COMP), a secreted glycoprotein synthesized by chondrocytes, regulates proliferation and type II collagen assembly. Mutations in the COMP gene cause pseudoachondroplasia and multiple epiphyseal dysplasia. Previously, we have shown that expression of D469del-COMP in transgenic mice causes intracellular retention of D469del-COMP, thereby recapitulating pseudoachondroplasia chondrocyte pathology. This inducible transgenic D469del-COMP mouse is the only in vivo model to replicate the critical cellular and clinical features of pseudoachondroplasia. Here, we report developmental studies of D469del-COMP-induced chondrocyte pathology from the prenatal period to adolescence. D469del-COMP retention was limited prenatally and did not negatively affect the growth plate until 3 weeks after birth. Results of immunostaining, transcriptome analysis, and qRT-PCR suggest a molecular model in which D469del-COMP triggers apoptosis during the first postnatal week. By 3 weeks (when most chondrocytes are retaining D469del-COMP), inflammation, oxidative stress, and DNA damage contribute to chondrocyte cell death by necroptosis. Importantly, by crossing the D469del-COMP mouse onto a Chop null background (Ddit3 null), thereby eliminating Chop, the unfolded protein response was disrupted, thus alleviating both D469del-COMP intracellular retention and premature chondrocyte cell death. Chop therefore plays a significant role in processes that mediate D469del-COMP retention. Taken together, these results suggest that there may be an optimal window before the induction of significant D469del-COMP retention during which endoplasmic reticulum stress could be targeted.
Assuntos
Acondroplasia/fisiopatologia , Condrócitos/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Glicoproteínas/metabolismo , Lâmina de Crescimento/patologia , Fator de Transcrição CHOP/fisiologia , Acondroplasia/embriologia , Animais , Apoptose , Proteínas Reguladoras de Apoptose/metabolismo , Desenvolvimento Ósseo/fisiologia , Cartilagem Articular/embriologia , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/metabolismo , Lâmina de Crescimento/embriologia , Lâmina de Crescimento/crescimento & desenvolvimento , Membro Posterior/embriologia , Membro Posterior/crescimento & desenvolvimento , Membro Posterior/metabolismo , Proteínas Matrilinas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Tíbia/embriologia , Tíbia/crescimento & desenvolvimento , Tíbia/metabolismo , Regulação para CimaRESUMO
Adult articular cartilage (AC) has a well described multizonal collagen structure. Knowledge of foetal AC organisation and development may provide a prototype for cartilage repair strategies, and improve understanding of structural changes in developmental diseases such as osteochondrosis (OC). The objective of this study was to describe normal development of the spatial architecture of the collagen network of equine AC using 1.5 T magnetic resonance imaging (MRI) and polarised light microscopy (PLM), at sites employed for cartilage repair studies or susceptible to OC. T2-weighted fast-spin echo (FSE) sequences and PLM assessment were performed on distal femoral epiphyses of equine foetuses, foals and adults. Both MRI and PLM revealed an early progressive collagen network zonal organisation of the femoral epiphyses, beginning at 4 months of gestation. PLM revealed that the collagen network of equine foetal AC prior to birth was already organised into an evident anisotropic layered structure that included the appearance of a dense tangential zone in the superficial AC in the youngest specimens, with the progressive development of an underlying transitional zone. A third, increasingly birefringent, radial layer developed in the AC from 6 months of gestation. Four laminae were observed on the MR images in the last third of gestation. These included not only the AC but also the superficial growth plate of the epiphysis. These findings provide novel data on normal equine foetal cartilage collagen development, and may serve as a template for cartilage repair studies in this species or a model for developmental studies of OC.