RESUMEN

The development of new technologies to produce three-dimensional and biocompatible scaffolds associated with high-end cell culture techniques have shown to be promising for the regeneration of tissues and organs. Some biomedical devices, as meniscus prosthesis, require high flexibility and tenacity and such features are found in polyurethanes which represent a promising alternative. The Poly(PCL-TMC)urethane here presented, combines the mechanical properties of PCL with the elasticity attributed by TMC and presents great potential as a cellular carrier in cartilage repair. Scanning electron microscopy showed the presence of interconnected pores in the three-dimensional structure of the material. The scaffolds were submitted to proliferation and cell differentiation assays by culturing mesenchymal stem cells in bioreactor. The tests were performed in dynamic flow mode at the rate of 0.4 mL/min. Laser scanning confocal microscopy analysis showed that the flow rate promoted cell growth and cartilage ECM synthesis of aggrecan and type II collagen within the Poly(PCL-TMC)urethane scaffolds. This study demonstrated the applicability of the polymer as a cellular carrier in tissue engineering, as well as the ECM was incremented only when under oriented flow rate stimuli. Therefore, our results may also provide data on how oriented flow rate in dynamic bioreactors culture can influence cell activity towards cartilage ECM synthesis even when specific molecular stimuli are not present. This work addresses new perspectives for future clinical applications in cartilage tissue engineering when the molecular factors resources could be scarce for assorted reasons.

Asunto(s)

Cartílago/química , Condrogénesis/efectos de los fármacos , Matriz Extracelular/química , Ingeniería de Tejidos , Reactores Biológicos , Cartílago/efectos de los fármacos , Cartílago/crecimiento & desarrollo , Cartílago/metabolismo , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Metacrilatos/química , Metacrilatos/farmacología , Poliésteres/química , Poliésteres/farmacología , Poliuretanos/química , Poliuretanos/farmacología , Andamios del Tejido/química

RESUMEN

BACKGROUND: Current clinical practice calls for pulse lavage of fresh osteochondral allografts (OCAs) to reduce immunogenicity; however, there is limited evidence of its effectiveness in reducing allogenic bone marrow elements. PURPOSE: To evaluate the effectiveness of pulse lavage in removing marrow elements from trabecular bone in fresh OCA transplantation. STUDY DESIGN: Controlled laboratory study. METHODS: The authors evaluated 48 fresh OCA plugs with 4 different common sizes (14- and 24-mm diameter, 6- and 10-mm thickness). Within each size group, half of the samples underwent pulse lavage (n = 6) with saline solution and half were left untreated (no lavage; control group, n = 6). For each treatment and size group, 3 samples were analyzed for DNA content as an indicator of the number of residual nucleated cells; the other 3 samples were histologically analyzed to assess the presence and distribution of cells within subchondral bone pores in 3 specific locations within the plug: peripheral, intermediate, and core. RESULTS: Osteochondral plugs treated with pulse lavage did not show a significant decrease in DNA content in comparison with untreated plugs. Overall, histological analysis did not show a significant difference between the treated and untreated groups (P = .23). Subgroup analysis by size demonstrated decreased marrow content in treated versus untreated groups in the thinner plug sizes (14 × 6 mm and 24 × 6 mm). Histological evaluation by zone demonstrated a significant difference between groups only in the peripheral zone (P = .04). CONCLUSION: Pulse lavage has limited effectiveness in removing marrow elements, in particular in plugs that are larger in diameter and, more importantly, in thickness. Better techniques for subchondral bone treatment are required for more thorough removal of potentially immunogenic marrow elements. CLINICAL RELEVANCE: OCA transplantation has become an established treatment modality. Unfortunately, OCA is not without limitations, chiefly its mode of failure through inadequate integration of the allograft subchondral bone with subsequent collapse. In an effort to improve integration, current clinical practice calls for pulse lavage to remove allogenic bone marrow from the subchondral bone in hopes of decreasing the immunogenicity of the graft and facilitating revascularization.

Asunto(s)

Aloinjertos/química , Médula Ósea/química , Huesos/química , Cartílago/química , ADN/análisis , Técnicas de Preparación Histocitológica/métodos , Irrigación Terapéutica , Trasplante Óseo , Huesos/anatomía & histología , Cartílago/anatomía & histología , Cartílago/trasplante , Humanos , Trasplante Homólogo

RESUMEN

Skate (Dipturus chilensis) cartilage extract was utilized as a green reducing agent for the synthesis of spherical gold nanoparticles with an average size of 16.7 ± 0.2 nm. The gold nanoparticle solution showed a surface plasmon resonance at 543 nm with a wine-red colour. A strong X-ray diffraction pattern and clear lattice structure in high-resolution transmission electron microscopy indicated a face-centred cubic structure of the gold nanoparticles. The gold nanoparticles retained excellent colloidal stability. Gold nanoparticles showed strong antioxidant activity in terms of 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. In vitro cytotoxicity was observed for seven cancer cells assessed by the water-soluble tetrazolium assay. Among the seven cancer cells, the highest cytotoxicity was observed for MDA-MB-231 (human breast adenocarcinoma cell) followed by HeLa (human epithelial cervix adenocarcinoma cell) and lastly by HT-29 (human colorectal adenocarcinoma cell). Furthermore, gold nanoparticles showed excellent haemocompatibility, indicating the possibility of their use as a future nanomedicine. These results strongly suggest that gold nanoparticles green-synthesized by upcycling skate cartilage waste extract will be valuable carriers or vehicles for the delivery of drugs or bioactive molecules, such as anti-cancer agents, for the treatment of cancers.

Asunto(s)

Cartílago/química , Oro/química , Oro/farmacología , Nanopartículas del Metal/química , Rajidae , Residuos , Aminoácidos/química , Animales , Antineoplásicos/síntesis química , Antineoplásicos/química , Antineoplásicos/farmacología , Antioxidantes/síntesis química , Antioxidantes/química , Antioxidantes/farmacología , Compuestos de Bifenilo/química , Técnicas de Química Sintética , Tecnología Química Verde , Células HT29 , Células HeLa , Humanos , Ensayo de Materiales , Picratos/química

RESUMEN

Chondroitin sulfate is a biomedical glycosaminoglycan (GAG) mostly used as a dietary supplement. We undertook analysis on some formulations of chondroitin sulfates available for oral administration. The analysis was based on agarose-gel electrophoresis, strong anion-exchange chromatography, digestibility with specific GAG lyases, uronic acid content, NMR spectroscopy, and size-exclusion chromatography. Keratan sulfate was detected in batches from shark cartilage, averaging ∼16% of the total GAG. Keratan sulfate is an inert material, and hazardous effects due to its presence in these formulations are unlikely to occur. However, its unexpected high percentage compromises the desired amounts of the real ingredient specified on the label claims, and forewarns the pharmacopeias to update their monographs. The techniques they recommended, especially cellulose acetate electrophoresis, are inefficient in detecting keratan sulfate in chondroitin sulfate formulations. In addition, this finding also alerts the manufacturers for improved isolation procedures as well as the supervisory agencies for better audits. Analysis based on strong anion-exchange chromatography is shown to be more reliable than the methods presently suggested by standard pharmacopeias.

Asunto(s)

Sulfatos de Condroitina/administración & dosificación , Sulfatos de Condroitina/química , Sulfato de Queratano/análisis , Administración Oral , Animales , Cartílago/química , Bovinos , Química Farmacéutica , Contaminación de Medicamentos , Humanos , Resonancia Magnética Nuclear Biomolecular , Tiburones , Extractos de Tejidos/química

RESUMEN

Glial fibrillary acidic protein (GFAP) is a member of the intermediary filament protein family. It is an important component of astrocytes and a known diagnostic marker of glial differentiation. GFAP is expressed in other neural tumors and pleomorphic adenoma and, less frequently, in cartilage tumors, chordomas, and soft tissue myoepitheliomas. The aim of this study was to evaluate the role of GFAP and its reliability in nonglial tumors as an immunohistochemical marker. We evaluated GFAP gene and protein expression using Q-PCR and immunohistochemistry, respectively, in 81 and 387 cases of soft tissue, bone tumors, and salivary pleomorphic adenomas. Immunohistochemistry staining for GFAP was observed in all osteosarcomas (8 cases), all pleomorphic adenomas (7 cases), in 5 of 6 soft tissue myoepitheliomas, and in 21 of 76 chondrosarcomas. By Q-PCR, GFAP was highly expressed in pleomorphic adenomas and, to a lesser extent, chondrosarcomas, soft tissue myoepitheliomas, and chondroblastic osteosarcomas. The results that we obtained by immunohistochemistry and Q-PCR were well correlated. GFAP is a potential marker for tumors with cartilaginous differentiation, supported by evidence that GFAP is expressed in certain cases of myoepithelial tumors by immunohistochemistry, including soft tissue myoepitheliomas, which are related to cartilaginous differentiation. These findings contribute significantly to the diagnosis of soft tissue myoepitheliomas with cartilaginous differentiation and chondroblastic osteosarcoma in mesenchymal tumors.

Asunto(s)

Adenoma Pleomórfico/química , Biomarcadores de Tumor/análisis , Neoplasias Óseas/química , Cartílago/química , Proteína Ácida Fibrilar de la Glía/análisis , Mioepitelioma/química , Neoplasias de las Glándulas Salivales/química , Neoplasias de los Tejidos Blandos/química , Adenoma Pleomórfico/genética , Adenoma Pleomórfico/patología , Biomarcadores de Tumor/genética , Neoplasias Óseas/genética , Neoplasias Óseas/patología , Cartílago/patología , Diferenciación Celular , Regulación Neoplásica de la Expresión Génica , Proteína Ácida Fibrilar de la Glía/genética , Humanos , Inmunohistoquímica , Mioepitelioma/genética , Mioepitelioma/patología , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias de las Glándulas Salivales/genética , Neoplasias de las Glándulas Salivales/patología , Neoplasias de los Tejidos Blandos/genética , Neoplasias de los Tejidos Blandos/patología

RESUMEN

Proteoglycans were accurately localized in mouse pubic symphyseal tissues using the cuprolinic blue method. Specific glycosaminoglycans degradative enzymes, together with chondroitin sulfate and decorin antibodies, allowed the identification of glycosaminoglycans. Chondroitin sulfate proteoglycans were the main proteoglycans observed in hyaline cartilage, fibrocartilage, and dense connective tissue. Ultrastructurally, they were seen as electron-dense granules and filaments. The granules, rich in chondroitin sulfate chains, were exclusively found in hyaline cartilage, whereas filaments were present in cartilage, fibrocartilage, and dense connective tissue. The latter were classified by size and susceptibility to enzyme digestion into F1, F2 and F3 filaments: F1 filaments were small, thin, and collagen fibril-associated; F2 filaments were thick, heavily stained, and localized around individual collagen fibrils and between bundles of collagen fibrils; and F3 filaments were scattered throughout elastic fiber surfaces. Considering their localization, susceptibility to chondroitinase AC and immunohistochemical detection, the symphysial F1 filaments were found to be preferentially decorin substituted with chondroitin sulfate side chains. The F2 filaments were also susceptible to chondroitinase AC treatment, whereas F3 filaments could be digested by heparitinase. The data thus obtained on the localization and identification of pubic symphyseal proteoglycans in virgin mice may be useful in the study of structural modifications that occur throughout pregnancy.

Asunto(s)

Proteoglicanos/análisis , Proteoglicanos/ultraestructura , Sínfisis Pubiana/química , Animales , Cartílago/química , Colorantes/química , Femenino , Inmunohistoquímica , Ratones , Microscopía Electrónica , Microscopía de Polarización/métodos , Sínfisis Pubiana/ultraestructura

RESUMEN

1. An experiment was designed to evaluate the effects of the addition of shark cartilage (SC) or chitosan (CH) to layer diets on egg component weights, yolk lipids and hen plasma lipids. 2. Hy-Line laying hens (80) were used during a 56 d feeding trial. Treatments were: basal diet (BD), BD + 20 g/kg SC, BD + 30 g/kg SC, BD + 20 g/kg CH and BD + 30 g/kg CH. Eggs were analysed on d 14, 28, 42 and 56. 3. Egg weight and egg component weights were not affected by these treatments throughout the experimental period. 4. After 14d of experimental feeding, cholesterol levels were higher in eggs from birds given BD + 20 g/kg CH and BD + 30 g/kg CH than in those from birds given BD. 5. Furthermore, eggs from hens given BD + 20 g/kg SC or BD + 20 g/kg CH were higher in palmitic and stearic acids and lower in oleic acid than those from birds fed on BD. After 56 d feeding, however, palmitic and stearic acid contents in eggs from hens given any of the supplemented diets were lower than in those from hens given BD, and oleic acid in eggs from hens given BD + 20 g/kg SC, BD + 30 g/kg SC and BD + 30 g/kg CH was higher than in those from birds fed on BD. 6. Plasma cholesterol and triacylglycerol levels were not significantly affected by dietary treatment. 7. Shark cartilage or chitosan at up to 30 g/kg in layer diets did not affect egg component weights (yolk, white and shell) and total lipid contents. During the period from 42 to 56d of experimental feeding, diets containing up to 30 g/kg chitosan reduced egg yolk contents of cholesterol, palmitic and stearic acids and increased the content of oleic acid.

Asunto(s)

Alimentación Animal , Cartílago/química , Pollos/fisiología , Quitina/análogos & derivados , Quitina/administración & dosificación , Yema de Huevo/química , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Cartílago/metabolismo , Pollos/metabolismo , Quitina/farmacología , Quitosano , Colesterol/análisis , Relación Dosis-Respuesta a Droga , Ácidos Grasos/análisis , Femenino , Lípidos/sangre , Oviposición/fisiología , Distribución Aleatoria , Tiburones

RESUMEN

The discovery that angiogenesis is a key condition for the growth of a tumor beyond a millimeter or two, brings about a new approach in the treatment of tumors using drugs able to inhibit the formation of new blood vessels. Also, it has been realized that antiangiogenic drugs can be useful in the treatment of other pathological processes, now classified as angiogenesis-dependent diseases. Initially, cartilage was considered as a possible natural source of antiangiogenic compounds due to its known avascular nature. To date, a number of in vitro and in vivo studies have suggested the existence of antiangiogenic and antitumor compounds in bovine and shark cartilage. However, the potential usefulness of shark cartilage in the treatment of cancer and other angiogenesis-dependent diseases have not been totally accepted due to (i) unsatisfactory patient outcome in clinical trials that have used shark cartilage in cancer patients, (ii) the lack of data that correlates bioavailability with pharmacological effects using oral shark cartilage. Thus, the objective of this review is to describe the main basic and clinical investigations reported in the literature, in which the antiangiogenic and/or antitumor properties of shark cartilage or of its extracts were evaluated. Possible explanations for conflicting results are discussed as well.

Asunto(s)

Inhibidores de la Angiogénesis/aislamiento & purificación , Inhibidores de la Angiogénesis/farmacología , Cartílago/química , Tiburones/metabolismo , Animales , Humanos

RESUMEN

The elastic tendon of the avian wing has been described by others as a unique structure with elastic properties due to the predominance of elastic fibers in the midsubstance. Further analyses of the tendon have shown it to possess five anatomically distinct regions. Besides the major elastic region, a distally located fibrocartilage and three tendinous regions are present. The tendinous regions connect: (1) the muscle to the elastic region, (2) the elastic region to the fibrocartilage and (3) the latter to the insertion site. The elastic region possesses thick and abundant elastic fibers and very thin, interconnecting collagen fibers. The collagen fibers in the sesamoid fibrocartilage are thick and interwoven, defining spaces occupied by fibrochondrocytes embedded in a non-fibrillar and highly metachromatic matrix. Biochemical analyses have shown that the fibrocartilage has about tenfold the amount of glycosaminoglycans (GAGs) found in the other regions. The main GAG in this region was chondroitin sulfate (CS) (plus keratan sulfate as detected immunocytochemically), while the other regions showed variable amounts of CS, dermatan sulfate (DS) and heparan sulfate. Further analyses have shown that a large CS-bearing proteoglycan is found in the fibrocartilage. The elastic region possesses two main proteoglycans, a large CS-bearing proteoglycan (which reacted with an antibody against keratan sulfate after chondroitinase ABC treatment) and a predominant DS-bearing proteoglycan, which showed immunoreactivity when assayed with an anti-biglycan antibody. The results demonstrate that the elastic tendon is a complex structure with complex regional structural and compositional adaptations, suited to different biomechanical roles.

Asunto(s)

Pollos/anatomía & histología , Tejido Elástico/anatomía & histología , Tejido Elástico/química , Proteoglicanos/análisis , Tendones/anatomía & histología , Tendones/química , Alas de Animales/anatomía & histología , Alas de Animales/química , Animales , Cartílago/anatomía & histología , Cartílago/química , Cartílago/ultraestructura , Colágeno/análisis , Tejido Elástico/ultraestructura , Glicosaminoglicanos/análisis , Glicosaminoglicanos/aislamiento & purificación , Histocitoquímica , Inmunohistoquímica , Microscopía Electrónica , Proteoglicanos/aislamiento & purificación , Tendones/ultraestructura , Alas de Animales/ultraestructura

RESUMEN

The effect of protein restriction on the growth of muscle, bone and body tissue of rats fed a diet containing 5% protein (experimental group) and 12% protein (control group) was studied. The following parameters were analyzed: body, muscle and cartilage growth, tibial length, plasma insulin IGF-1 concentrations, and tissue proteoglycan, protein, RNA and insulin-like growth factor-1 concentrations. Protein synthesis in tissues was also determined. Except for protein concentration in the two tissues and of insulin-like growth factor-1 in cartilage, all other values differed significantly between groups. The lower ration and protein ingestion by group 1 receiving a 5% case in diet provoked a reduced secretion of the anabolic hormones insulin and IGF-1 and a lower synthesis of proteoglycans, RNA and protein, impairing total body growth and growth of the tissues analyzed.

Asunto(s)

Desarrollo Óseo/fisiología , Factor I del Crecimiento Similar a la Insulina/fisiología , Desarrollo de Músculos , Animales , Peso Corporal/fisiología , Desarrollo Óseo/efectos de los fármacos , Cartílago/química , Cartílago/metabolismo , Caseínas/administración & dosificación , Ingestión de Alimentos/efectos de los fármacos , Músculo Esquelético/química , Músculo Esquelético/metabolismo , Músculos/efectos de los fármacos , Deficiencia de Proteína/fisiopatología , Proteínas/farmacología , Ratas , Ratas Wistar

RESUMEN

There is overwhelming evidence to indicate that free radicals cause oxidative damage to lipids, proteins and nucleic acids and are involved in the pathogenesis of several degenerative diseases. Therefore, antioxidants, which can neutralize free radicals, may be of central importance in the prevention of these disease states. The protection that fruits and vegetables provide against disease has been attributed to the various antioxidants contained in them. Recently, an anti-inflammatory and analgesic activity of a water-soluble fraction from shark cartilage has been described. Using electrophoretical assays, bacteria survival and transformation and the Salmonella/mammalian-microsome assay, we investigated the putative role of shark cartilage-containing preparation in protecting cells against reactive oxygen species induced DNA damage and mutagenesis. If antimutagens are to have any impact on human disease, it is essential that they are specifically directed against the most common mutagens in daily life. Our data suggest that shark cartilage-containing preparation can play a scavenger role for reactive oxygen species and protects cells against inactivation and mutagenesis.

Asunto(s)

Cartílago/química , Depuradores de Radicales Libres/farmacología , Mutágenos/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Tiburones/metabolismo , 4-Nitroquinolina-1-Óxido/toxicidad , Animales , Electroforesis en Gel de Agar , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Radicales Libres/metabolismo , Peróxido de Hidrógeno/toxicidad , Pruebas de Mutagenicidad , Plásmidos/efectos de los fármacos , Plásmidos/genética , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Compuestos de Estaño/toxicidad

RESUMEN

The present work shows an antinociceptive and dose-dependent effect of shark cartilage hydrosoluble fraction (HF) on writhing and formalin tests in mice. The effect was not altered by thalidomide, a known inhibitor of tumor necrosis factor-alfa (TNF-alfa) synthesis. Similarly, the antinociceptive effect did not change in the presence of naloxone, indicating that the opioid system is not involved. However, the effect observed was blocked by L-arginine, a NO synthesis substrate, and it was potentiated by L-NAME, suggesting a role of the NO system in the shark cartilage antinociceptive effect. Effects similar to those seen with the HF were detected with peak II from gel filtration chromatography. The increase in vascular permeability induced by serotonin in rats was significantly abolished by the HF at the dose of 2 mg/kg, p.o., and again it was not potentiated by thalidomide. The observed blockade in the vascular permeability increase induced by histamine was detected only with a higher dose (10 mg/kg, p.o.).

Asunto(s)

Analgésicos/farmacología , Antiinflamatorios no Esteroideos/farmacología , Cartílago/química , Óxido Nítrico/metabolismo , Analgésicos/química , Analgésicos/metabolismo , Animales , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/metabolismo , Permeabilidad Capilar/efectos de los fármacos , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Interacciones Farmacológicas , Inhibidores Enzimáticos/farmacología , Histamina/farmacología , Masculino , Ratones , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Ratas , Ratas Wistar , Serotonina/farmacología , Tiburones

RESUMEN

Natural products from flora and fauna are frequently used as nutritional supplements and medicaments. Two short-term assays were carried out and negative results were obtained for shark-cartilage containing preparation. The tests employed were the Salmonella/mammalian microsome assay using tester strains TA97, TA98, TA100, TA102 and TA1535 with or without S9 mix and the SOS-Chromotest with Escherichia coli strain PQ37. Evidence for shark-cartilage containing preparation functioning as an antimutagen was detected. Using bacterial survival assays with Escherichia coli fpg (BH20) and xthA (BW9091), we investigated the putative role of shark-cartilage containing preparation in protecting cells against lesions induced by hydrogen peroxide in normal and low iron level conditions. Our data suggest that shark-cartilage containing preparation can play a scavenger role for reactive oxygen species and protect against DNA lesions in both conditions.

Asunto(s)

Antimutagênicos/farmacología , Cartílago/química , Peróxido de Hidrógeno/toxicidad , 2,2'-Dipiridil/farmacología , 4-Nitroquinolina-1-Óxido/metabolismo , 4-Nitroquinolina-1-Óxido/toxicidad , Animales , Antioxidantes/farmacología , Biotransformación , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Depuradores de Radicales Libres/farmacología , Peróxido de Hidrógeno/metabolismo , Hierro/metabolismo , Mutagénesis , Pruebas de Mutagenicidad , Mutágenos/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Respuesta SOS en Genética , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Tiburones

RESUMEN

1. The effect of biomechanical forces on large proteoglycans and collagen of cartilage has deserved intensive study, enhancing the importance of these molecules to support a better distribution of compressive forces especially in articular cartilage. In the present study, other extracellular matrix components, non-collagenous proteins and small proteoglycans, have been evaluated in terms of biomechanical tension. 2. Different parts of chicken xiphoid cartilage, lateral (R and L) and central (C) portions, which bear different biomechanical tensions, were analyzed. DEAE-cellulose chromatography profiles were similar for R and L portions. SDS-PAGE analyses revealed proteins of 29, 60 and 70 kDa for R and L. The 20- and 70-kDa proteins were not detected in the C portion while the 60-kDa protein was present at a high level. 3. The differences found between lateral (R and L) and central portions of the xiphoid cartilage may be related to the structure of the cartilage which bears higher tension forces than the lateral parts.

Asunto(s)

Cartílago/química , Proteoglicanos/aislamiento & purificación , Animales , Fenómenos Biomecánicos , Pollos , Matriz Extracelular/química , Matriz Extracelular/fisiología , Proteoglicanos/fisiología , Apófisis Xifoides

RESUMEN

1. Two proteoglycans, PG1 and PG2, have been isolated from shark cartilage. Both are highly polydisperse and large (molecular mass: 1-10 x 10(6) Daltons) and contain chondroitin sulfate and keratan sulfate side chains, but PG2 is somewhat smaller than PG1 and contains less keratan sulfate. 2. Monoclonal antibodies were raised against PG1. Many antibodies were obtained and one of them, MST1, was subcloned and further characterized. This monoclonal antibody reacts with PG1 and PG2 from shark cartilage and also with aggrecan from bovine trachea cartilage. Chondroitinase AC-treated proteoglycans react with MST1, indicating that the antibody does not recognize chondroitin sulfate. MST1 also recognizes aggrecan from human cartilage and a proteoglycan from bovine brain (neurocan) but it does not recognize proteoglycans from rat Walker tumor, fetal calf muscle and decorin from human myoma. 3. Using MST1 we were able to demonstrate that both PG1 and PG2 aggregate with hyaluronic acid.

Asunto(s)

Anticuerpos Monoclonales/aislamiento & purificación , Cartílago/química , Proteoglicanos/química , Animales , Bovinos , Sulfatos de Condroitina/química , Sulfatos de Condroitina/inmunología , Sulfatos de Condroitina/aislamiento & purificación , Epítopos , Humanos , Sulfato de Queratano/química , Sulfato de Queratano/inmunología , Sulfato de Queratano/aislamiento & purificación , Masculino , Ratones , Ratones Endogámicos BALB C , Proteoglicanos/inmunología , Proteoglicanos/aislamiento & purificación , Conejos , Ratas , Tiburones

RESUMEN

1. Two proteoglycans, PG1 and PG2, have been isolated from shark cartilage. Both are highly polydisperse and large (molecular mass: 1-10 x 10**6 Daltons) and contain chondroitin sulfate and keratan sulfate side chains, but PG2 is somewhat smaller tham PG1 and contains less keratan sulfate. 2. Monoclonal antibodies were raised against PG1. Many antibodies were obtained and one of them, MST1, was subcloned and furter characterized. This monoclonal antibody reacts with PG1 and PG2 from shark cartilage and also with aggrecan from bovine trachea cartilage. Chondroitinase AC-treated proteglycans react MST1, indicating that the antibody does not reconize chondroitin sulfate. MST1 also recognizes aggrecan from human cartilage and a proteoglycan from bovine brain (neurocan) but it does reconize proteoglycans from rat Walker tumor, fetal calf muscle and decorin from human myoma. 3. Using MST1 we were able to demonstrate that both PG1 aggregate with hyaluronic acid

Asunto(s)

Bovinos , Ratones , Conejos , Ratas , Humanos , Animales , Masculino , Anticuerpos Monoclonales/aislamiento & purificación , Cartílago/química , Proteoglicanos/química , Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/inmunología , Condroitinsulfatasas/química , Condroitinsulfatasas/inmunología , Condroitinsulfatasas/aislamiento & purificación , Epítopos , Sulfato de Queratano/química , Sulfato de Queratano/inmunología , Sulfato de Queratano/aislamiento & purificación , Proteoglicanos/inmunología , Proteoglicanos/aislamiento & purificación

RESUMEN

1. The effect of biomechanical forces on larges proteglycans and collagen of cartilage has deserved intensive study, enhacing the importance of these molecules to support a better distribution of compressive forces especially in articular cartilage. In the present study, other extracellular matrix components, non-collagenous proteins and small proteglycans, have been evaluated in terms of biomechanical tension. 2. Different parts of chicken xiphoid cartilage, lateral (R and L) and central (C) portions, which bear different biomechanical tensions, were analyzed. DEAE-cellulose chromatography profiles were similar for R and L portions. SDS-PAGE analyses revealed proteins of 29, 60 and 70 KDa for R and L. The 20-and 70-KDa proteins were not detected in the C portion while the 60-KDaprotein was presented at a high level. 3. The differences found between lateral (R and L) and central portions of the xiphoid cartilage may be related to the struycture of the cartilage which bears higher tension forces than the lateral parts

Asunto(s)

Animales , Cartílago/química , Proteoglicanos/aislamiento & purificación , Fenómenos Biomecánicos , Pollos , Cromatografía DEAE-Celulosa , Electroforesis en Gel de Poliacrilamida , Matriz Extracelular/química , Matriz Extracelular/fisiología , Proteoglicanos/fisiología

RESUMEN

In order to evaluate the effect of chaotropic agents on proteoglycan and non-collagenous proteins, chicken xiphoid cartilage was treated with guanidine-HCI and MgCl2 in different concentrations (1M to 5M), and different periods of time (12, 24, 48 and 72hr). The maximum yield of uronic acid was obtained with 3M MgCl2 (73.3 per cent). Concentrations of 4M and 5M of MgCl2 showed that much less uronic acid was removed, 55.3 per cent and 38.1 respectively. Extraction with 3M MgCl2 and 3M guanidine-HCl resulted better efficiency when performed for 48 hr. Analysis by SDS-PAGE of the extracts obtained with guanidine-HCl and MgCl, in different concentrations pointed out that most components are equally removed with the two solvents, showing that the extraction with MgCl2 is an alternative assay to remove non-collagenous proteins from extracellular matrix

Asunto(s)

Animales , Cartílago/química , Cloruro de Magnesio/farmacología , Guanidinas/farmacología , Proteínas de la Matriz Extracelular/aislamiento & purificación , Pollos , Electroforesis en Gel de Poliacrilamida

RESUMEN

In order to evaluate the effect of chaotropic agents on proteoglycan and non-collagenous proteins, chicken xiphoid cartilage was treated with guanidine-HCl and MgCl2 in different concentrations (1M to 5M), and different periods of time (12, 24, 48 and 72 hr). The maximum yield of uronic acid was obtained with 3M MgCl2 (73.3%). Concentrations of 4M and 5M of MgCl2 showed that much less uronic acid was removed, 55.3% and 38.1% respectively. Extraction with 3M MgCl2 and 3M guanidine-HCl resulted better efficiency when performed for 48 hr. Analysis by SDS-PAGE of the extracts obtained with guanidine-HCl and MgCl2 in different concentrations pointed out that most components are equally removed with the two solvents, showing that the extraction with MgCl2 is an alternative assay to remove non-collagenous proteins from extracellular matrix.

Asunto(s)

Cartílago/química , Proteínas de la Matriz Extracelular/aislamiento & purificación , Guanidinas/farmacología , Cloruro de Magnesio/farmacología , Animales , Pollos , Electroforesis en Gel de Poliacrilamida , Guanidina

RESUMEN

We investigated the immunohistochemical presence of various collagen types in bone and cartilage tissue from an infant Peruvian mummy dating between 500 and 1000 A.D. which had been excavated at the necropolis of Las Trancas in the Nazca region in Peru. Following careful rehydration and decalcification of the tissue, the mummy tissue showed morphologically good preservation of the matrix, which could be shown to be composed of various collagen types in a typical pattern. Bone consisted of a collagen I matrix with a small rim of collagen III and V at the endosteal lining and a pericellular collagen V staining around osteocytic holes. In the hypertrophic cartilage of the epiphyseal growth plate, a typical pattern of collagen types II and X could be found. These observations provide evidence that in well-preserved mummy tissue the antigenic determinants of major matrix components are still adequately preserved for an immunohistochemical analysis. This technique may thus be a very helpful tool for the analysis of pathologic processes of historic bone tissue. It may also allow in certain circumstances a distinction between pseudopathologic tissue destruction and pathologic tissue alteration.

Asunto(s)

Huesos/química , Cartílago/química , Colágeno/análisis , Momias , Femenino , Historia Antigua , Humanos , Inmunohistoquímica , Lactante , Perú