RESUMEN
BACKGROUND AND AIMS: A prospective long-term follow-up study of bioactive glass (BAG)-S53P4 and autogenous bone (AB) used as bone graft substitutes for posterolateral spondylodesis in treatment of unstable lumbar spine burst fractures during 1996-1998 was conducted. MATERIAL AND METHODS: The lumbar fractures were fixed using posterior USS instrumentation. BAG was implanted on the left side of the fusion-bed and AB on the right side. The operative outcome was evaluated on X-rays and CT scans, and a clinical examination was also performed. RESULTS: The Oswestry score was excellent, and the mean pain score 1. The mean compression rate of the injured vertebral body was 25%. A solid bony fusion was seen on CT scans on the AB side in all patients and on the BAG side in five patients, and a partial fusion in five patients, resulting in a total fusion-rate of 71% of all fused segments in the BAG group. CONCLUSIONS: Our long-term results show that BAG-S54P4 bone graft material is safe to be used as a bone graft extender in spine surgery.
Asunto(s)
Sustitutos de Huesos/uso terapéutico , Vértebras Lumbares/lesiones , Fracturas de la Columna Vertebral/cirugía , Fusión Vertebral/métodos , Adulto , Trasplante Óseo , Femenino , Estudios de Seguimiento , Vidrio , Humanos , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Estudios Prospectivos , Trasplante AutólogoRESUMEN
The functions of the intervertebral disc and of articular cartilage are intimately related to their aggrecan content. Aggrecan is a proteoglycan that interacts with hyaluronan to form large aggregates, which are responsible for the ability of the tissues to resist compressive loads. This function is related to the structure of aggrecan, and in particular to the large number of chondroitin sulphate chains present on its core protein. The chondroitin sulphate chains are present in two adjacent regions of the aggrecan core protein, termed the CS1 and CS2 domains. In the human, the region of the aggrecan gene encoding the CS1 domain exhibits size polymorphism, which can result in variation in the degree of chondroitin sulphate substitution of aggrecan in different individuals. This raises the possibility that the functional properties of aggrecan may vary between individuals, and that those individuals with an inferior aggrecan structure may be more susceptible to premature intervertebral disc or articular cartilage degeneration. Several studies have been performed to demonstrate such an association, but the results have been ambiguous. This review explains the relationship between aggrecan structure and function, describes the technique used to assess aggrecan polymorphism and the conclusions and limitations of the data obtained to date, and discusses the implications for tissue degeneration and clinical practice.
Asunto(s)
Cartílago Articular/fisiopatología , Proteoglicanos Tipo Condroitín Sulfato/genética , Proteoglicanos Tipo Condroitín Sulfato/fisiología , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/fisiología , Disco Intervertebral/fisiopatología , Lectinas Tipo C/genética , Lectinas Tipo C/fisiología , Osteoartritis/fisiopatología , Polimorfismo Genético , Enfermedades de la Columna Vertebral/fisiopatología , Agrecanos , Alelos , Proteoglicanos Tipo Condroitín Sulfato/química , Sulfatos de Condroitina/fisiología , ADN/genética , Electroforesis en Gel de Agar , Proteínas de la Matriz Extracelular/química , Regulación de la Expresión Génica , Humanos , Ácido Hialurónico/fisiología , Lectinas Tipo C/química , Osteoartritis/etiología , Osteoartritis/genética , Reacción en Cadena de la Polimerasa , Estructura Terciaria de Proteína , Enfermedades de la Columna Vertebral/genéticaRESUMEN
The present study was carried out to determine whether immobilization-induced (Im) osteopenic bone possesses the same reparative capacity as normal healthy bone. Furthermore, the effects of mechanical loading versus immobilization on bone defect healing were studied. Three-week cast-immobilization was used to induce local osteopenia in mice. A standardized metaphyseal bone defect of the distal femur was created unilaterally both in immobilization-induced (Im) osteopenic mice and in nonimmobilized (Mo) age-matched control animals. After creation of the bone defect, the animals in both groups were further divided into two groups: 3-week cast-immobilization (Im-Im and Mo-Im) groups, and unrestricted weight-bearing (Im-Mo and Mo-Mo) groups. The healing process was followed up to 3 weeks using RNA analysis, histomorphometry, biomechanical testing, and pQCT measurements. At 3 weeks of healing without immobilization, bone mineral density (BMD), as well as bone bending stiffness and strength were higher in normal (Mo-Mo) than in osteopenic (Im-Mo) bone. Although the levels of mRNAs characteristic to chondrocytes (Sox9 and type II collagen), hypertrophic chondrocytes (Type X collagen), osteoblasts (type I collagen and osteocalcin), and osteoclasts (cathepsin K) during the bone defect healing exhibited similarities in their expression profiles, mechanical loading conditions also caused characteristic differences. Mechanical loading during healing (Mo-Mo group) induced stronger expression of cartilage- and bone-specific genes and resulted in higher BMD than that seen in the cast-immobilized group (Mo-Im). In biomechanical analysis, increased bending stiffness and strength were also observed in animals that were allowed weight-bearing during healing. Thus, our study shows that bone healing follows the same molecular pathway both in osteopenic and normal bones and presents evidence for reduced or delayed regeneration of noncritical size defects in immobilization-induced osteopenic bone.
Asunto(s)
Enfermedades Óseas Metabólicas/fisiopatología , Regeneración Ósea , Fémur/fisiopatología , Animales , Fenómenos Biomecánicos , Densidad Ósea , Enfermedades Óseas Metabólicas/genética , Femenino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , ARN/genéticaRESUMEN
This study is based on a hypothesis that overexpression of an osteoclast enzyme, cathepsin K, causes an imbalance in bone remodeling toward bone loss. The hypothesis was tested in transgenic (TG) mice harboring additional copies of the murine cathepsin K gene (Ctsk) identifiable by a silent mutation engineered into the construct. For this study, three TG mouse lines harboring 3-25 copies of the transgene were selected. Tissue specificity of transgene expression was determined by Northern analysis, which revealed up to 6-fold increases in the levels of cathepsin K messenger RNA (mRNA) in calvarial and long bone samples of the three TG lines. No changes were seen in the mRNA levels of other osteoclast enzymes, indicating that the increase in cathepsin K mRNA was not a reflection of activation of all osteoclast enzymes. Immunohistochemistry confirmed that cathepsin K expression in the TG mice was confined to osteoclasts and chondroclasts. Histomorphometry revealed a significantly decreased trabecular bone volume (BV), but, surprisingly, also a marked increase in the number of osteoblasts, the rate of bone turnover, and the amount of mineralizing surface (MS). However, monitoring of bone density in the proximal tibias of the TG mice with peripheral quantitative computed tomography (pQCT) failed to reveal statistically significant changes in bone density. Similarly, no statistically significant alterations were observed in biomechanical testing at the age of 7 months. The increases in parameters of bone formation triggered by increased cathepsin K expression is an example of the tight coupling of bone resorption and formation during the bone-remodeling cycle.
Asunto(s)
Enfermedades Óseas Metabólicas/metabolismo , Remodelación Ósea/fisiología , Catepsinas/fisiología , Animales , Densidad Ósea , Enfermedades Óseas Metabólicas/etiología , Catepsina K , Catepsinas/genética , Catepsinas/metabolismo , Fémur/metabolismo , Fémur/patología , Expresión Génica , Humanos , Inmunoquímica/métodos , Ratones , Ratones Transgénicos , Osteoblastos/metabolismo , Osteoclastos/metabolismo , ARN Mensajero/metabolismo , Cráneo/metabolismo , Cráneo/patología , Distribución TisularRESUMEN
A bone defect model was developed in the distal metaphysis of the femur for studies on bone healing in the mouse. The circular defect involving 20% of the bone circumference resulted in a 34% reduction in the bending moment compared to intact bone. The healing process was followed using histomorphometry, peripheral quantitative computed tomography (pQCT), biomechanical testing, and molecular biological analyses. Histologically, healing of the defect was characterized by filling of the medullary cavity with trabecular new bone during the first week of healing, and by closing of the cortical window by 6 weeks. Small areas of periosteal chondrogenesis were frequently observed during defect healing. In pQCT, bone mineral content (BMC) of the defect area approached that of intact control bone already by 3 weeks, reflecting the production of trabecular bone. Similarly, the bending strength and stiffness of the healing femur reached the level of intact control femur already at 3 weeks. Bone formation and remodeling was followed by Northern analyses, which demonstrated elevated mRNA levels for bone components (type I collagen and osteocalcin), and for osteoclastic enzymes (cathepsin K, matrix metalloproteinase-9, and tartrate-resistant acid phosphatase) throughout the healing period. Finally, the applicability of the defect model for gene therapy experiments was tested using adenovirus-mediated transfer of the LacZ reporter gene. Both histochemistry and mRNA analyses demonstrated that the gene was expressed in the repair tissue with the highest expression during the first week of healing. The present model thus provides a standardized environment for studies on induction and remodeling of trabecular new bone in normal and genetically engineered mice.
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Fracturas del Fémur/patología , Curación de Fractura , Modelos Animales , Animales , Fenómenos Biomecánicos , Northern Blotting , Femenino , Fracturas del Fémur/diagnóstico por imagen , Fracturas del Fémur/fisiopatología , Técnicas de Transferencia de Gen , Inmunohistoquímica , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/genética , ARN Mensajero/metabolismo , Tomografía Computarizada por Rayos XRESUMEN
An experimental mouse model for disuse osteopenia was developed using unilateral cast immobilization. Analysis of the distal femurs and proximal tibias by quantitative histomorphometry revealed significant osteopenia within 10-21 days of immobilization. At 3 weeks, bone loss was also demonstrated with peripheral quantitative computed tomography as diminished bone mineral content and as concomitant reduction in the cross-sectional moment of inertia. These structural and geometrical alterations resulted in decreased strength of the distal femurs tested by cantilever bending. Analysis of the underlying cellular and molecular mechanisms of bone loss revealed a rapid increase in bone resorption within 3 days of immobilization. The mRNA levels for cathepsin K, matrix metalloproteinase-9, and tartrate resistant acid phosphatase were all significantly increased during the 21-day immobilization period, but with different expression profiles. These increases were paralleled by an increased number of osteoclasts as measured by histomorphometry. By day 6 of immobilization, the balance of bone turnover was further shifted toward net bone loss as the mRNA levels for major bone components (type I collagen and osteocalcin) were decreased. In histomorphometric analysis this was observed as reduced rates of mineral apposition and bone formation after 10 days of immobilization. The results of this study demonstrate that immobilization has a dual negative effect on bone turnover involving both depressed bone formation and enhanced bone resorption.
Asunto(s)
Enfermedades Óseas Metabólicas/metabolismo , Osteoblastos/metabolismo , Animales , Biomarcadores , Northern Blotting , Densidad Ósea , Enfermedades Óseas Metabólicas/etiología , Enfermedades Óseas Metabólicas/genética , Catepsinas/genética , Decorina , Modelos Animales de Enfermedad , Proteínas de la Matriz Extracelular , Femenino , Fémur/patología , Perfilación de la Expresión Génica , Inmovilización , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Osteocalcina/genética , Procolágeno/genética , Proteoglicanos/genética , ARN Mensajero , Tibia/patologíaRESUMEN
BACKGROUND AND AIMS: This study was carried out to investigate the use of porous bioactive glass implants in promotion of articular cartilage and subchondral bone repair in large osteochondral joint defects. MATERIAL AND METHODS: Two conical osteochondral defects (top diameter 3.0-3.2 mm) were drilled into the patellar grooves of the distal femurs in the rabbit. The defects, extending (approximately 6-7 mm) from the surface of the articular cartilage to the subchondral marrow space, were reconstructed with size-matched porous conical implants made of sintered bioactive glass microspheres (microsphere diameter 250-300 microm, structural implant compression strength 20-25 MPa) using press-fit technique. The implant surface was smoothened to the level of the surrounding articular cartilage. One of the two defects in each femur was left empty to heal naturally and to serve as the control. At 8 weeks, the defect healing was analyzed with use of a semiquantitative histological grading system, histomorphometry of subchondral bone repair, back-scattered electron imaging of scanning electron microscopy (BEI-SEM), and a microindentation test for characterization for the stiffness properties of the cartilage repair tissue. RESULTS: The porous structure of the bioactive glass implants, extending from the articular defect of the patellar groove into the posterior cortex of the femur, was extensively filled by new bone. Cartilage repair varied from near-complete healing by hyaline cartilage to incomplete healing predominantly by fibrocartilage or fibrous tissue. There were, however, no statistical differences in the histological scores of repair between the glass-filled and control defects, although the sum of the averages of each category was lowest for the bioactive glass filled defects. The indentation stiffness values of all the defects were also significantly lower than that of normal cartilage on the patellar groove. CONCLUSIONS: Porous textures made by sintering bioactive glass microspheres may expand the opportunities in reconstruction of deep osteochondral defects of weight-bearing joints. The implants act mechanically as a supporting scaffold and facilitate the penetration of stromal bone marrow cells and their chondrogenic and osteogenic differentiation. Ionic properties of the bioactive glasses make the substances highly potential even as delivery systems for adjunct growth factor therapy.
Asunto(s)
Sustitutos de Huesos , Cartílago Articular/cirugía , Fémur/cirugía , Vidrio , Animales , Materiales Biocompatibles , Microscopía Electrónica de Rastreo , Microesferas , Procedimientos Ortopédicos , Oseointegración , Conejos , Dióxido de SilicioRESUMEN
In preparation for gene manipulative experiments in mice we have isolated genomic clones covering the entire murine cathepsin K gene (Ctsk). Sequence analysis revealed that the gene spans 10.1 kb and contains eight exons and seven introns. The sizes of the coding exons 2-7 as well as the pattern of intron sizes are conserved between the human and mouse cathepsin K genes. Genomic organization of the mouse cathepsin K gene also closely resembles those of cathepsins S and L. More than 26% of the Ctsk sequence consists of different repetitive elements. Detailed sequence analysis of 1.7 kb of Ctsk promoter revealed several putative binding sites for transcription factors and two stretches of 280-320 bp which were > 70% homologous with the human cathepsin K promoter. Analysis of genomic clones extending further upstream of the Ctsk gene identified the 3' end of the gene coding for arylhydrocarbon-receptor nuclear translocator (Arnt). This places Ctsk approximately 4.5 kb downstream of Arnt on mouse chromosome 3 at locus 47.9.
Asunto(s)
Catepsinas/genética , Mapeo Cromosómico , Proteínas de Unión al ADN , Receptores de Hidrocarburo de Aril , Factores de Transcripción/genética , Regiones no Traducidas 5' , Animales , Translocador Nuclear del Receptor de Aril Hidrocarburo , Secuencia de Bases , Catepsina K , Variación Genética , Humanos , Intrones , Ratones , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADNRESUMEN
We have constructed cDNA clones covering the entire coding region of mouse, human and rabbit preprocathepsin K mRNA for studies on bone turnover. The clone pMCatK-1 for mouse cathepsin K shares 87% nucleotide homology with the corresponding human and rabbit sequences. Analysis of a panel of mouse tissues for tissue distribution of cathepsin K mRNA revealed the highest levels in musculoskeletal tissues: bone, cartilage and skeletal muscle. In situ hybridization of developing mouse embryos was performed to identify the cellular source of cathepsin K mRNA. The strongest mRNA signal was detected in osteoclasts of bone, identified in serial sections by positive TRAP staining. Cathepsin K mRNA was also observed in some hypertrophic chondrocytes of growth cartilages. Association of cathepsin K production with degradation of bone and cartilage matrix suggests that this enzyme and its mRNA levels could serve as markers for matrix degradation in diseases affecting these tissues.