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1.
Med Mol Morphol ; 57(1): 76-81, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38071257

RESUMO

In this study, we investigated specific and characteristic findings of the surface layer of surgical resected disc specimens in human temporomandibular joint osteoarthritis cases by transmission electron microscopy (TEM).Specimens were surgically removed from the TMJ of 5 cases (4 female patients: 5 cases) clinically osteoarthritis. Following findings were observed by TEM. Images were photographed on a JEM1400-Flash Electron microscope (JEOL, Japan) equipped with an EM-14661FLASH high-sensitivity digital complementary metal-oxide-semiconductor camera.Following findings were observed by TEM. 1) The surface is covered with plump fibroblastic and histiocytoid cells. 2) Collagen fiber bundles and collagenous matrix are exposed onto the eroded disc surface. 3) Fibrinous dense material is observed on the eroded disc surface. 4) Bundles of collagen fibers are densely observed. 5) Collagen bundles are rich around capillary vessels. 6) Synovial surface cells reveal features of activated macrophages with vacuole formation. Especially, plump fibroblastic and histiocytoid cells, and activated macrophages with vacuole, which were significant findings of the surface layer. These findings might have a significant effect on the regulation of synovial fluid.


Assuntos
Osteoartrite , Transtornos da Articulação Temporomandibular , Humanos , Feminino , Elétrons , Membrana Sinovial/ultraestrutura , Articulação Temporomandibular/cirurgia , Microscopia Eletrônica de Transmissão , Colágeno/ultraestrutura
2.
J Equine Vet Sci ; 133: 104988, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38157949

RESUMO

The purpose of the study was to determine the histo-morphological effects on villous synovium after synovectomy using two different motorized synovial resectors and two different intensities ex-vivo. Thirty-three (n = 33) equine metacarpophalangeal/metatarsophalangeal joints were used. Synovectomy was performed along the dorsomedial/dorsolateral synovium (n = 66) using two motorized synovial resectors (aggressive full radius resector, AFRR, used at two intensities: single treatment, n = 24 vs. triple treatment, n = 21 vs. aggressive meniscus side cutter, AMSC, n = 21). Arthroscopic images were evaluated blindly for resector type and intensity. Histological images were evaluated descriptive for synovial morphology and the extent of tissue loss using a microscopic scale. Scanning electron microscopy described the synovial morphology. The synovectomized areas were specific for each resector used and distinguishable from arthroscopic images. The AFRR demonstrated a clear demarcation between treated and non-treated areas and removed the stratum synoviale completely including parts of the underlying stratum fibrosum. In contrast, the AMSC showed less clear demarcation, villous scaffolds and no involvement of the stratum fibrosum. Triple intense treated AFFR samples resulted in significantly deeper lesions compared to single treatments (p = 0.037) but could not be distinguished on arthroscopic images. The morphological effects on villous synovium differ according to the resector type used. The extent of synovial tissue loss cannot be estimated from arthroscopic images but histologically. The type and use of motorized synovial resector determines the morphological alterations of the treated synovium. Arthroscopic control is considered unsuitable to control synovectomy depth.


Assuntos
Artroscopia , Sinovectomia , Cavalos , Animais , Sinovectomia/veterinária , Artroscopia/métodos , Artroscopia/veterinária , Membrana Sinovial/patologia , Membrana Sinovial/ultraestrutura
3.
Nat Immunol ; 22(12): 1551-1562, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34811544

RESUMO

Misdirected immunity gives rise to the autoimmune tissue inflammation of rheumatoid arthritis, in which excess production of the cytokine tumor necrosis factor (TNF) is a central pathogenic event. Mechanisms underlying the breakdown of self-tolerance are unclear, but T cells in the arthritic joint have a distinctive metabolic signature of ATPlo acetyl-CoAhi proinflammatory effector cells. Here we show that a deficiency in the production of mitochondrial aspartate is an important abnormality in these autoimmune T cells. Shortage of mitochondrial aspartate disrupted the regeneration of the metabolic cofactor nicotinamide adenine dinucleotide, causing ADP deribosylation of the endoplasmic reticulum (ER) sensor GRP78/BiP. As a result, ribosome-rich ER membranes expanded, promoting co-translational translocation and enhanced biogenesis of transmembrane TNF. ERrich T cells were the predominant TNF producers in the arthritic joint. Transfer of intact mitochondria into T cells, as well as supplementation of exogenous aspartate, rescued the mitochondria-instructed expansion of ER membranes and suppressed TNF release and rheumatoid tissue inflammation.


Assuntos
Artrite Reumatoide/metabolismo , Ácido Aspártico/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Mitocôndrias/metabolismo , Membrana Sinovial/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , ADP-Ribosilação , Transferência Adotiva , Animais , Artrite Reumatoide/genética , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Autoimunidade , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/transplante , Linfócitos T CD4-Positivos/ultraestrutura , Estudos de Casos e Controles , Células Cultivadas , Retículo Endoplasmático/imunologia , Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/ultraestrutura , Chaperona BiP do Retículo Endoplasmático/metabolismo , Feminino , Humanos , Masculino , Camundongos , Mitocôndrias/imunologia , Mitocôndrias/transplante , Mitocôndrias/ultraestrutura , Membrana Sinovial/imunologia , Membrana Sinovial/ultraestrutura , Fator de Necrose Tumoral alfa/genética
4.
Lasers Med Sci ; 36(3): 529-540, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32519204

RESUMO

The presence of intra-articular crystals is detected in different articular pathologies of acute or chronic nature. The aim of this work was to analyze the action of the indium gallium aluminum and phosphorus (InGaAlP) (λ = 670 nm) laser on the synovial membrane present in the knee joint in experimentally induced microcrystalline arthritis in male adult Wistar rats. The animals were divided into three experimental groups (n = 24): control (A), experimentally induced arthritis (B), experimentally induced arthritis+InGaAlP laser therapy (C). The laser treatment was made daily in the patellar region of the right knee after 48 h of the experimental induction. After 7, 14, and 21 days of therapy, the rats were euthanized and the right knees were removed and processed for histomorphometric, immunohistochemical, ultrastructural, and biochemical investigation of the synovium. The number of granulocytes on the 14th and 21st days was higher in B and lower in C and, lastly, in A. The number of fibroblasts on the 14th and 21st days was similar between A and C and below B. The number of blood vessels on the 21st day was higher in B than in the other groups. The positive number of cells for the TUNEL test was higher on the 14th and 21st days in B compared to the others. The percentage of tissue area occupied by birefringent collagen fibers was higher in B on the 21st day than in the others. The ultrastructure of cells showed fibroblast-like morphology in all groups and periods evaluated. The quantification of glycosaminoglycans did not present significant differences between the groups in all the experimental periods. The amount of hydroxyproline was higher in B compared to the other groups on the 14th and 21st days. The content of non-collagen proteins was higher in B on the 21st day in relation to the other groups. Quantification of TNF-α on the 21st day was higher in A and B than in C. For TGF-ß on the 21st day, groups B and C presented similar and higher values than A. For MMP-13, groups A and B presented data similar to and above C. In relation to ADAMT-S4, on the 21st day, groups B and C presented data similar to and lower than A. InGaAlP-670 nm therapy reduced the inflammatory process and tissue injuries of the synovial membrane in comparison to the untreated group, indicating its potential utilization in clinical studies aiming in the recovery of acute arthritis in patients.


Assuntos
Artrite Experimental/cirurgia , Terapia a Laser , Membrana Sinovial/patologia , Membrana Sinovial/efeitos da radiação , Proteína ADAMTS4/metabolismo , Animais , Apoptose/efeitos da radiação , Vasos Sanguíneos/patologia , Vasos Sanguíneos/efeitos da radiação , Cristalização , Articulação do Joelho/patologia , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Ratos Wistar , Membrana Sinovial/ultraestrutura , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
5.
Microscopy (Oxf) ; 70(2): 186-191, 2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-32744639

RESUMO

Since the majority of a tendon's dry weight is collagen fibers, tendon healing consists mainly of collagen repair and observing three-dimensional networks of collagen fibers with scanning electron microscopy (SEM) is optimal for investigating this process. In this report, a cell-maceration/SEM method was used to investigate extrasynovial tendon (unwrapped tendon in synovial tissue such as the tendon sheath) healing of an injured Achilles tendon in a rat model. In addition, since mechanical stimulation is important for tendon healing, a novel, tensionless, rat lower leg tendon injury model was established and verified by visualizing the structural change of collagen fibers under tensionless conditions by SEM. This new model was created by transplanting the leg of a rat with a tendon laceration to the back, removing mechanical stimulation. We then compared the process of tendon healing with and without tension using SEM. Under tension, collagen at the tendon stump shows axial alignment and repair that subsequently demarcates the paratenon (connective tissue on the surface of an extrasynovial tendon) border. In contrast, under tensionless conditions, the collagen remains randomly arranged. Our findings demonstrate that mechanical stimulation contributes to axial arrangement and reinforces the importance of tendon tension in wound healing.


Assuntos
Tendão do Calcâneo/lesões , Tendão do Calcâneo/ultraestrutura , Microscopia Eletrônica de Varredura/métodos , Estresse Mecânico , Traumatismos dos Tendões/diagnóstico por imagem , Animais , Colágeno/metabolismo , Colágeno/ultraestrutura , Imageamento Tridimensional/métodos , Modelos Animais , Ratos , Membrana Sinovial/ultraestrutura , Cicatrização/fisiologia
6.
J Orthop Surg Res ; 15(1): 97, 2020 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-32151262

RESUMO

OBJECTIVE: The aim of this study is to compare the efficiency of different separation techniques for extracting synovial tissue-derived exosomes. METHODS: The synovial tissue discarded during knee arthroscopy or total knee arthroplasty surgery was collected from the Third Affiliated Hospital of Beijing University of Chinese Medicine. Ultracentrifugation (UC), filtration combined with size exclusion chromatography (SECF), and 8% polyethylene glycol (PEG) were used to extract synovial tissue-derived exosomes. Transmission electron microscopy (TEM), nanoparticle tracer analysis (NTA), and Western Blot (WB) were used to detect the morphology, particle size, and biomarker proteins (CD9, CD63, Flotillin-1, and calnexin) of exosomes. RESULTS: The extracts of enriched round and discoid vesicles were successfully extracted with UC, SECF, and PEG. The results of TEM have shown that all three extraction methods can extract circular or elliptical vesicles with disc- and cup-shaped structures from the synovial tissue, with the diameter is about 30-150 nm. NTA suggested the main peaks of three groups of exosomes are around 100-120 nm, and the concentration of the three groups of exosomes was greater than 1 × 1010/ml. The results of WB showed that three positive protein markers (CD9, CD63, and Flotillin-1) were highly expressed in the suspension extracted by the three methods and low in the synovial tissue. However, the negative protein (calnexin) was highly expressed in synovial tissues and PEG group, while low in UC and SECF group. CONCLUSION: Morphology, particle size, and labeled protein marker detection confirmed that UC, SECF, and PEG can extract exosomes derived from synovial tissue; UC and SECF are more recommended for the extraction of synovial tissue-derived exosomes, which provides a methodological basis for studying the function and mechanism of synovial tissue exosomes in the future.


Assuntos
Western Blotting/métodos , Cromatografia em Gel/métodos , Exossomos/ultraestrutura , Microscopia Eletrônica de Transmissão/métodos , Membrana Sinovial/ultraestrutura , Ultracentrifugação/métodos , Humanos , Nanopartículas/análise , Nanopartículas/ultraestrutura , Tamanho da Partícula , Membrana Sinovial/química , Membrana Sinovial/citologia
7.
BMC Anesthesiol ; 18(1): 51, 2018 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-29743011

RESUMO

BACKGROUND: Intra-articular local anaesthetics are widely used for providing postoperative analgesia and decreasing the need for opioids. Procaine has proven positive effects in carpal tunnel syndrome and chondromalacia patella. However, the effect of procaine on articular cartilage has not yet been studied. The aim of this study was to evaluate the effects of intra-articular procaine injection on the articular cartilage and the synovium. METHODS: Twenty adult Sprague-Dawley rats were enrolled in the study. After providing anaesthesia and aseptic conditions, 0.25 ml of 10% procaine was injected to the right knee joint, and 0.25 ml of normal saline (as control group) was injected to the left knee joint. Knee joint samples were obtained from four rats in each group after appropriate euthanasia on days 1, 2, 7, 14 and 21. The histological sections of the articular and periarticular regions and the synovium were evaluated by two histologists, and inflammatory changes were graded according to a five-point scale in a blinded manner. The apoptosis of chondrocytes was determined by the caspase-3 indirect immunoperoxidase method. RESULTS: There were no significant differences in inflammation between procaine and saline groups at any of the time intervals. Slight inflammatory infiltration due to injection was seen in both groups on the 1st day. Haemorrhage was observed in both groups at days 1 and 2, and the difference between groups was not found to be significant. No significant difference was detected in the percentage of apoptotic chondrocytes between groups at any of the time intervals. CONCLUSIONS: Injection of procaine seems safe to use intra-articularly based on this in vivo study on rat knee cartilage. However, further studies investigating both the analgesic and histopathological effects of procaine on damaged articular cartilage and synovium models are needed.


Assuntos
Anestésicos Locais/farmacologia , Cartilagem Articular/efeitos dos fármacos , Membro Posterior/efeitos dos fármacos , Procaína/farmacologia , Solução Salina/farmacologia , Membrana Sinovial/efeitos dos fármacos , Animais , Cartilagem Articular/ultraestrutura , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Masculino , Ratos , Ratos Sprague-Dawley , Membrana Sinovial/ultraestrutura
8.
Sci Rep ; 8(1): 3581, 2018 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-29483562

RESUMO

A new cell type named telocyte (i.e. cell with distinctive prolongations called telopodes) has recently been identified in the stroma of various organs in humans. However, no study has yet reported the existence of telocytes in the synovial membrane of diarthrodial joints. This work was therefore undertaken to search for telocytes in the normal human synovium using transmission electron microscopy, immunohistochemistry and immunofluorescence. Ultrastructural analyses demonstrated the presence of numerous spindle-shaped telocytes in the whole synovial sublining layer. Synovial telocytes exhibited very long and thin moniliform telopodes and were particularly concentrated at the boundary between the lining and sublining layers and around blood vessels. Light microscopy confirmed the presence of CD34-positive telocytes in the aforementioned locations. Moreover, synovial telocytes coexpressed CD34 and platelet-derived growth factor receptor α. Double immunostaining further allowed to unequivocally differentiate synovial telocytes (CD34-positive/CD31-negative) from vascular endothelial cells (CD34-positive/CD31-positive). The in vitro examination of fibroblast-like synoviocyte primary cultures revealed the coexistence of different cell types, including CD34-positive telocytes projecting typical moniliform telopodes. In conclusion, our work provides the first evidence that telocytes do exist in the human synovium and lays the groundwork for future studies on synovial telocytes in a variety of degenerative and destructive joint diseases.


Assuntos
Membrana Sinovial/citologia , Membrana Sinovial/ultraestrutura , Telócitos/metabolismo , Telócitos/ultraestrutura , Adulto , Antígenos CD34/metabolismo , Células Cultivadas , Células Endoteliais/metabolismo , Imunofluorescência , Hospitais Universitários , Humanos , Imuno-Histoquímica , Itália , Masculino , Microscopia Eletrônica de Transmissão , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/metabolismo , Sinoviócitos/ultraestrutura , Telopódios
9.
Cytokine ; 96: 1-7, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28267648

RESUMO

Rheumatoid arthritis (RA) is an autoimmune disease which may lead to severe disabilities due to structural joint damage and extraarticular manifestations The dendritic cell marker CD83 belongs to the immunoglobulin superfamily and has previously been associated with autoimmune diseases. In RA the levels of soluble CD83 (sCD83) are elevated in synovial fluid, however little is known about CD83 expression and regulation in RA. Therefore, we studied how CD83 is expressed in RA and further evaluated the effect of anti-TNF-α therapy hereon. Early RA patients were randomized to conventional disease modifying anti-rheumatic drugs with or without additional anti-TNF-α therapy. Rheumatoid arthritis patients had increased levels of sCD83 in plasma compared with healthy volunteers. The increase in sCD83 plasma levels were unaffected by anti-TNF-α therapy. In chronic RA patients the levels of sCD83 were higher in synovial fluid than in plasma, and only a limited amount of membrane bound CD83 expression was detected on the surface of cells from peripheral blood and synovial fluid. Finally, confocal microscopy of RA synovial membranes revealed that CD83 was mainly localized intracellularly in a group of cells with diverse morphology including both antigen-presenting cells and non-antigen-presenting cells. Our findings demonstrate that early-stage RA patients have elevated levels of sCD83 in plasma and that anti-TNF-α treatment has no effect on the sCD83 plasma level. This suggest that in RA patients sCD83 regulation is beyond control of TNF-α.


Assuntos
Adalimumab/uso terapêutico , Antígenos CD/sangue , Antígenos CD/genética , Artrite Reumatoide/sangue , Imunoglobulinas/sangue , Imunoglobulinas/genética , Glicoproteínas de Membrana/sangue , Glicoproteínas de Membrana/genética , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adalimumab/administração & dosagem , Adalimumab/efeitos adversos , Biomarcadores/sangue , Células Dendríticas/imunologia , Feminino , Humanos , Imunoterapia , Inflamação/terapia , Masculino , Metotrexato/administração & dosagem , Metotrexato/uso terapêutico , Microscopia Confocal , Pessoa de Meia-Idade , Líquido Sinovial/química , Líquido Sinovial/imunologia , Membrana Sinovial/ultraestrutura
10.
Am J Vet Res ; 77(12): 1332-1339, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27901386

RESUMO

OBJECTIVE To evaluate 4 methods for generating decellularized equine synovial extracellular matrix. SAMPLE Villous synovium harvested from the femoropatellar and medial femorotibial joints of 4 healthy adult horses < 7 years of age. Synovial samples were frozen (-80°C) until used. PROCEDURES Synovial samples were thawed and left untreated (control) or decellularized with 1 of 4 methods (15 samples/horse/method): incubation in 0.1% peracetic acid (PAA), incubation in 0.1% PAA twice, incubation in 1% Triton X-100 followed by incubation in DNase, and incubation in 2M NaCl followed by incubation in DNase. Control and decellularized samples were examined for residual cells, villous integrity, and collagen structure and integrity by means of histologic examination and scanning electron microscopy; cell viability was evaluated by means of culture and exclusion staining. Decellularization efficiency was assessed by testing for DNA content and DNA fragment size. RESULTS Incubation in PAA once preserved the synovial villous architecture, but resulted in high DNA content and retention of large (> 25,000 base pair) DNA fragments. Incubation in Triton and incubation in NaCl resulted in low DNA content and short (< 200 base pair) DNA fragments, but destroyed the synovial villous architecture. Incubation in PAA twice resulted in low DNA content and short DNA fragments while retaining the synovial villous architecture. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that of the methods evaluated, incubation in 0.1% PAA twice was the best method for generating decellularized equine synovial extracellular matrix.


Assuntos
Matriz Extracelular , Membrana Sinovial/citologia , Tecidos Suporte/veterinária , Animais , Bioengenharia , Colágeno , Cavalos , Microscopia Eletrônica de Varredura/veterinária , Joelho de Quadrúpedes , Membrana Sinovial/ultraestrutura , Engenharia Tecidual
11.
Eur Cell Mater ; 31: 407-24, 2016 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-27232667

RESUMO

Many researchers world over are currently investigating the suitability of stromal cells harvested from foetal tissues for allogeneic cell transplantation therapies or for tissue engineering purposes. In this study, we have investigated the chondrogenic potential of mesenchymal stromal cells (MSCs) isolated from whole sections of human umbilical cord or mixed cord (UCSCs-MC), and compared them with cells isolated from synovial membrane (SMSCs), Hoffa's fat pad (HFPSCs) and cartilage. All MSCs were positive for surface markers including CD73, CD90, CD105, CD44, CD146 and CD166, but negative for CD11b, CD19, CD34, CD45 and HLA-DR in addition to CD106 and CD271. Chondrogenic potential of all cell sources was studied using 3D pellet cultures incubated in the presence of different combinations of anabolic substances such as dexamethasone, IGF-1, TGF-ß1, TGF-ß3, BMP-2 and BMP-7. BMP-2 and dexamethasone in combination with TGF-ß1 or TGF-ß3 excelled at inducing chondrogenesis on SMSCs, HFPSCs and chondrocytes, as measured by glycosaminoglycans and collagen type II staining of pellets, quantitative glycosaminoglycan expression, quantitative PCR of cartilage signature genes and electron microscopy. In contrast, none of the tested growth factor combinations was sufficient to induce chondrogenesis on UCSCs-MC. Moreover, incubation of UCSCs-MC spheroids in the presence of cartilage pieces or synovial cells in co-cultures did not aid chondrogenic induction. In summary, we show that in comparison with MSCs harvested from adult joint tissues, UCSCs-MC display poor chondrogenic abilities. This observation should alert researchers at the time of considering UCSCs-MC as cartilage forming cells in tissue engineering or repair strategies.


Assuntos
Técnicas de Cultura de Células/métodos , Condrogênese , Células-Tronco Mesenquimais/citologia , Tecidos Suporte/química , Cordão Umbilical/citologia , Tecido Adiposo/citologia , Tecido Adiposo/ultraestrutura , Cartilagem/citologia , Proliferação de Células , Separação Celular , Técnicas de Cocultura , DNA/metabolismo , Citometria de Fluxo , Regulação da Expressão Gênica , Glicosaminoglicanos/metabolismo , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/ultraestrutura , Pessoa de Meia-Idade , Fenótipo , Esferoides Celulares/citologia , Esferoides Celulares/metabolismo , Células Estromais/citologia , Membrana Sinovial/citologia , Membrana Sinovial/ultraestrutura
12.
Eur J Pharm Biopharm ; 96: 388-95, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26369477

RESUMO

Viscosupplementation (VS) is a therapy for osteoarthrosis (OA) consisting of repetitive intra-articular injections of hyaluronic acid (HA). It is known to be clinically effective in relieving pain and increasing joint mobility by restoring joint homeostasis. In this study, the effects of two novel HA-based VS hydrogel formulations were assessed and challenged against a pure HA commercial formulation for the first time and this in a rabbit model of early OA induced by anterior cruciate ligament transection (ACLT). The first formulation tested was a hybrid hydrogel composed of HA and reacetylated chitosan, a biopolymer considered to be chondroprotective, assembled thanks to an ionic shielding. The second formulation consisted of a novel HA polymer grafted with antioxidant molecules (HA-4AR) aiming at decreasing OA oxidative stress and increasing HA retention time in the articulation. ACLT was performed on rabbits in order to cause structural changes comparable to traumatic osteoarthrosis. The protective effects of the different formulations were observed on the early phase of the pathology in a full randomized and blinded manner. The cartilage, synovial membrane, and subchondral bone were evaluated by complementary investigation techniques such as gross morphological scoring, scanning electron microscopy, histological scoring, and micro-computed tomography were used. In this study, ACLT was proven to successfully reproduce early OA articular characteristics found in humans. HA and HA-4AR hydrogels were found to be moderately protective for cartilage as highlighted by µCT. The HA-4AR was the only formulation able to decrease synovial membrane hypertrophy occurring in OA. Finally, the hybrid HA-reacetylated chitosan hydrogel surprisingly led to increased subchondral bone remodeling and cartilage defect formation. This study shows significant effects of two innovative HA modification strategies in an OA rabbit model, which warrant further studies toward more effective viscosupplementation formulations.


Assuntos
Antioxidantes/uso terapêutico , Modelos Animais de Doenças , Ácido Hialurônico/uso terapêutico , Articulação do Joelho/efeitos dos fármacos , Osteoartrite do Joelho/tratamento farmacológico , Resorcinóis/uso terapêutico , Viscossuplementos/uso terapêutico , Acetilação , Animais , Antioxidantes/efeitos adversos , Antioxidantes/química , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Osso e Ossos/ultraestrutura , Cartilagem/efeitos dos fármacos , Cartilagem/metabolismo , Cartilagem/patologia , Cartilagem/ultraestrutura , Química Farmacêutica , Quitosana/efeitos adversos , Quitosana/química , Quitosana/farmacocinética , Quitosana/uso terapêutico , Preparações de Ação Retardada/efeitos adversos , Preparações de Ação Retardada/química , Preparações de Ação Retardada/uso terapêutico , Liberação Controlada de Fármacos , Ácido Hialurônico/efeitos adversos , Ácido Hialurônico/química , Hidrogéis , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Articulação do Joelho/ultraestrutura , Masculino , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Coelhos , Distribuição Aleatória , Resorcinóis/efeitos adversos , Resorcinóis/química , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia , Membrana Sinovial/ultraestrutura , Viscossuplementos/efeitos adversos , Viscossuplementos/química
13.
Microsc Res Tech ; 78(6): 479-84, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25807922

RESUMO

To illustrate the mechanism of lymphatic reabsorption in knee joint effusion. The current investigation employed transmission electron microscopy (TEM) and scanning electron microscopy (SEM) techniques to reveal the ultrastructure of the knee synovial membrane in New Zealand rabbits and human. Ultrastructural changes of the synovial lymphatic stomata were observed by using trypan blue absorption and sodium hydroxide (NaOH) digestion methods, and the animal models of synovitis. New Zealand rabbits and human synovial membranes were composed of two types of synovial cells: type A and type B. No lymphatic stomata were found among type A synovial cells, whereas lymphatic stomata with the diameters ranging 0.74-3.26 µm were found in type B synovial cells, and some stomata were closed. After the NaOH digestion, a number of sieve pores, similar to lymphatic stomata in size and shape, were observed in the dense fibrous connective tissue underneath the type B synovial cells. After injecting trypan blue into the rabbit knee joint cavity, absorption of trypan blue through the lymphatic stomata was observed, suggesting the absorption function of the synovial lymphatic stomata. In the rabbit knee joint synovitis models, the synovial lymphatic stomata diameter enlarged. Some macrophages migrated from the lymphatic stomata, indicating that the synovial lymphatic stomata were involved in the joint effusion absorption and inflammatory response. Our study is the first to report the existence of synovial lymphatic stomata in the New Zealand rabbits and human knee joints. Lymphatic stomata may have an important role in the reabsorption of joint effusion.


Assuntos
Líquido Ascítico/metabolismo , Articulação do Joelho/ultraestrutura , Estomas Peritoneais/ultraestrutura , Membrana Sinovial/ultraestrutura , Sinovite/patologia , Animais , Transporte Biológico , Humanos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Coelhos
14.
J Forensic Sci ; 59(6): 1575-82, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25228058

RESUMO

Articular cartilage was examined to determine its decomposition sequence and its potential for assessing the postmortem interval. Scanning electron microscopy of articular cartilage from buried porcine trotters showed the presence of microcrystals on the synovial surface. These orthorhombic pyramidal or "coffin"-shaped crystals, appeared at 3 weeks (22 days) after interment and disappeared after 6 weeks. The disappearance of these crystals was linked to decompositional changes to the integrity of the synovial joint. The formation and disappearance of these crystals was associated with a pH change at the cartilage surface. Scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) analysis showed that the five main elements contained within these crystals were carbon, nitrogen, oxygen, magnesium, and phosphorous. Such elemental analysis suggested the crystals may be struvite (MgNH4 PO4 6(H2 O)). Bacteria cultured from the cartilage synovial surface produced struvite crystals when grown in suitable media and were identified by DNA analysis to be Comamonas sp.


Assuntos
Cartilagem Articular/química , Cartilagem Articular/ultraestrutura , Cristalização , Mudanças Depois da Morte , Animais , Carbono/análise , Comamonas/isolamento & purificação , Meios de Cultura , Patologia Legal , Concentração de Íons de Hidrogênio , Magnésio/análise , Compostos de Magnésio , Microscopia Eletrônica de Varredura , Modelos Animais , Nitrogênio/análise , Oxigênio/análise , Fosfatos , Fósforo/análise , Espectrometria por Raios X , Estruvita , Suínos , Membrana Sinovial/ultraestrutura
15.
Pathol Res Pract ; 210(10): 694-7, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25023880

RESUMO

We report a morphologically rare type of tenosynovial giant cell tumor (TSGCT), localized type, occurring in a 49-year-old man. Imaging examination revealed multiple nodular lesions around the right knee joint. The largest one extended to both intra- and extra-osseous region of the right distal femur. Histologically, the tumor consisted of epithelioid mononuclear cells and they looked like to have abundant eosinophilic cytoplasm reminiscent of hepatic tissues. In some areas, however, typical histologic features of TSGCT were observed. Electron microscopy revealed that the eosinophilic cytoplasm-like substance was intercellular fibrinous material surrounding the mononuclear tumor cells. Immunohistochemically, mononuclear tumor cells and multinucleate giant cells were positive for CD68 (Kp1) and some of the mononuclear tumor cells were also positive for desmin. Finally, we made the diagnosis of hepatoid TSGCT.


Assuntos
Tumores de Células Gigantes/patologia , Neoplasias de Tecidos Moles/patologia , Membrana Sinovial/ultraestrutura , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Biópsia/métodos , Tumores de Células Gigantes/diagnóstico , Humanos , Masculino , Microscopia Eletrônica/métodos , Pessoa de Meia-Idade , Neoplasias de Tecidos Moles/diagnóstico
16.
J Comput Assist Tomogr ; 38(4): 503-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24733006

RESUMO

OBJECTIVE: The purposes of our study were to analyze magnetic resonance imaging (MRI) and cadaveric findings concerning the medial synovial fold of the posterior cruciate ligament (PCL) and to classify the types of fold according to anatomic location. METHODS: Two musculoskeletal radiologists reviewed MR images of 17 cadaveric knees to classify the types of medial fold of the PCL by consensus. The MRI types were divided into 3 groups. In type A, there was no definitive medial fold; and in type B, inferior-short type, there was a small protrusion of the medial border. Type C, inferior-long type, had a long enough fold to exceed the imaginary line, which is connecting between the medial tibial condyle and posterolateral aspect of the medial femoral condyle. Correlations were sought between the findings derived from the MRI studies and cadaveric dissections. Histologic analyses of the medial fold were also performed. RESULTS: On MRI, the most common type of medial fold was type B (76.4%), followed by type C (11.8%) and type A (11.8%). In the cadaveric investigation, the medial folds of both types B and C were found to project into the medial femorotibial joint. Moreover, there was also a protruding medial fold at the superior aspect of the PCL in the A. Histologic examination of the medial folds revealed collagenous tissue surrounded by synovial cells. CONCLUSIONS: Medial folds of the PCL are normal synovial structures that can be seen by MRI and in cadaveric studies in a large proportion of the population.


Assuntos
Articulação do Joelho/anatomia & histologia , Imageamento por Ressonância Magnética/métodos , Ligamento Cruzado Posterior/anatomia & histologia , Membrana Sinovial/anatomia & histologia , Idoso , Idoso de 80 Anos ou mais , Cadáver , Feminino , Humanos , Articulação do Joelho/ultraestrutura , Masculino , Microscopia de Polarização/métodos , Pessoa de Meia-Idade , Variações Dependentes do Observador , Ligamento Cruzado Posterior/ultraestrutura , Membrana Sinovial/ultraestrutura
17.
Int J Immunopathol Pharmacol ; 26(4): 917-30, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24355227

RESUMO

The association between microfracture of the subchondral plate and a coverage scaffold has emerged as a promising strategy to treat cartilage lesions in a one-step procedure. Between different types of scaffolds (e.g. collagen, hyaluronic acid, polyglycolic acid) currently studied, type I collagen scaffold is the most used for this purpose, and is currently adopted for humans. The aim of this study was to test a novel scaffold made of mixed type I and II collagen (I-IICS) in order to define the immunological reaction of the synovial tissue and the repair capabilities induced by the collagen membrane when associated with microfracture. Eight New Zealand White rabbits, aged 180 days, were operated on bilaterally on the medial femoral condyle. A circular cartilage lesion was performed up to the calcified layer of the medial femoral condyle, and the centre of the lesion was microfractured. Randomly, one of the two lesions was covered with the I-IICS (treated), and the other was left uncovered (control). The synovial membrane reaction and the quality of the cartilage tissue repair were investigated at 2, 90, 180 and 270 days macroscopically, histomorphologically and ultrastructurally. Expression of tumor necrosis factor-alpha (TNF-alpha) in synovial tissue by immunocytochemistry analyses was also investigated. In the control group, at 2 days gold particles were localized mainly on synoviocyte type A, less on synoviocytes type B and on collagen bundles; in the treated group the reaction is more intense in cells in the matrix, but at 180 days controls and treated joints were very similar. The synovial membranes of the joints receiving the I-IICS did not reveal significant changes compared to the age-matched controls. Signs of inflammation were present at the 90-day time-point, and became less evident at afterwards. The degradation of the scaffolds was already evident at the 90-day time-point. The quality of the cartilage repair of the rabbits treated with the I-IICS was slightly better in 5 cases out of 6 in comparison to the controls. However, a statistically significant difference was not detected (p=0.06). Scaffolds made of mixed type I and II collagen exhibited good biocompatibility properties in vivo and favoured cartilage restoration when associated with microfracture, as shown in this pilot study.


Assuntos
Cartilagem/cirurgia , Colágeno Tipo II/farmacologia , Colágeno Tipo I/farmacologia , Membrana Sinovial/ultraestrutura , Tecidos Suporte , Animais , Projetos Piloto , Coelhos , Cicatrização
18.
Morfologiia ; 144(5): 50-3, 2013.
Artigo em Russo | MEDLINE | ID: mdl-24592718

RESUMO

The structural and functional reorganization of the synovial membrane of the knee joint capsule was studied in 22 individuals of mature age (30-65 years) of both genders with the use of light and electron microscopy, histochemistry, immunohistochemistry and morphometry. The morpho-functional peculiarities of the synovial membrane remodeling in the II period of mature age included: increase the in the synovial intima thickness; significant accumulation of type IV collagen at the border of synovial intima with the subintimal fibrovascular layer of synovial membrane; activation of matrix metalloproteinases MMP-2 and MMP-9; increased cell proapoptotic activity in synovial intima. Collectively, these changes may initiate the disturbances in fluid production and the development of the degenerative-dystrophic processes in the articular cartilage at this stage of ontogenesis.


Assuntos
Imuno-Histoquímica , Articulação do Joelho/ultraestrutura , Microscopia Eletrônica , Membrana Sinovial/ultraestrutura , Adulto , Idoso , Envelhecimento , Autopsia , Colágeno/metabolismo , Feminino , Humanos , Articulação do Joelho/metabolismo , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , Membrana Sinovial/metabolismo , Adulto Jovem
19.
Microscopy (Oxf) ; 62(2): 317-20, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23081991

RESUMO

To evaluate the advantages of combination of two advanced electron microscopic technologies such as serial block-face scanning electron microscopy and double-axis electron beam tomography, we analyzed the three-dimensional morphology of cellular relationships between dendritic and plasma cells in the synovial membrane from patients with rheumatoid arthritis, using the combined approach.


Assuntos
Artrite Reumatoide/patologia , Células Dendríticas/ultraestrutura , Plasmócitos/ultraestrutura , Membrana Sinovial/ultraestrutura , Comunicação Celular , Células Dendríticas/fisiologia , Humanos , Processamento de Imagem Assistida por Computador , Imageamento Tridimensional , Microscopia Eletrônica de Varredura , Plasmócitos/fisiologia , Tomografia Computadorizada por Raios X
20.
Ultrastruct Pathol ; 36(4): 207-18, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22849522

RESUMO

INTRODUCTION: Few ultrastructural studies have been reported in psoriatic arthritis (PsA). The authors report a series of synovial biopsies with emphasis on patients with early disease to look for distinctive light (LM) and electron microscopic (EM) features of possible importance. METHODS: The authors examined synovial biopsies obtained primarily by needle biopsy from 13 PsA patients using LM and/or EM. Sections from 12 patients were evaluated by LM for vascularity, synovial lining thickness, fibrin deposition, and inflammation via a semi-quantitative scale. Nine EM specimens were descriptively analyzed. Clinical, synovial fluid (SF), and radiographic characteristics were recorded. RESULTS: Patients were mostly male, with mean disease duration before biopsy of 2.19 ± 2.60 years; 7 patients had arthritis for less than 1 year. All patients had peripheral arthritis, 2 had axial involvement. SFs disclosed predominance of polymorphonuclear leukocytes. LM demonstrated proliferation of synovial lining cells, lymphocyte and plasma cell infiltration, as well as dramatic clusters of small vessels in the superficial synovium. EMs showed more detailed vascular changes, including small, subendothelial, electron-dense deposits and scattered microparticles in vessel lumens and walls. CONCLUSIONS: Prominent vascularity is confirmed as an important feature of some PsA. Vascular changes and other features, including the first EM demonstration of microparticles in PsA (identified as potent factors in other inflammatory joint diseases), are potential targets for therapy.


Assuntos
Artrite Psoriásica/patologia , Membrana Sinovial/patologia , Sinovite/patologia , Adulto , Idoso , Biópsia , Feminino , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Neutrófilos/patologia , Neutrófilos/ultraestrutura , Membrana Sinovial/ultraestrutura , Adulto Jovem
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