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1.
Arch Orthop Trauma Surg ; 139(11): 1607-1615, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31363834

RESUMEN

PURPOSE: To clinically evaluate an arthroscopic autologous chondrocyte implantation (ACI) technique with an in situ crosslinking matrix for the treatment of full thickness cartilage defects of the knee and to present histological results of a graft cartilage biopsy obtained after 1.5 years. METHODS: Fifteen cases of arthroscopic autologous chondrocyte implantation in the knee performed between November 2011 and October 2012 were included in the study. Medical charts and operational reports were screened and the patients were contacted after 0.8 ± 0.3 years (0.4-1.3) and 4.3 ± 0.3 years (4.0-4.8) to asses subjective IKDC and re-operation. The Tegner activity scale was collected at the second follow-up time point. Subjective IKDC response rates were assessed at both follow-up time points. RESULTS: The first and second follow-up was completed by all 15 patients (100%). The subjective IKDC scores showed a significant improvement (pre-operative 44.5 ± 15.9, first follow-up 71.1 ± 15.9, p < 0.001, second follow-up 72.6 ± 17.3, p < 0.001). The overall response rate was 66.7% (n = 10) at follow-up one and two. There were no significant differences in pre-injury (4, range 1-9) and follow-up two (4, range 2-7) Tegner activity scales (p = n.s.). Two patients required re-operation in the index knee, not related to the ACI procedure. No complication related to the ACI or the implantation technique occurred. The histological results showed excellent cartilage regeneration. CONCLUSION: Arthroscopic ACI using an in situ crosslinking matrix is a safe and reliable treatment option for full-thickness cartilage defects of the knee.


Asunto(s)
Artroscopía/métodos , Condrocitos/trasplante , Articulación de la Rodilla , Trasplante Autólogo/métodos , Enfermedades de los Cartílagos/cirugía , Humanos , Articulación de la Rodilla/citología , Articulación de la Rodilla/cirugía
2.
Artif Cells Nanomed Biotechnol ; 47(1): 3188-3193, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31366242

RESUMEN

Objective: To investigate the effects of miR-15a on proliferation and apoptosis of human knee articular chondrocytes and explore its underlying mechanism. Methods: qRT-PCR was used to detect the expression of miR-15a in normal chondrocytes and knee arthritic chondrocytes; miR-con (transfected miR-con), miR-15a (transfected miR-15a mimics), anti-miR-con group (transfected anti-miR-con), anti-miR-15a group (transfected anti-miR-15a mimics), pcDNA group (transfected pcDNA), pcDNA-SMAD2 group (transfected pcDNA-SMAD2), the miR-15a + pcDNA group (co-transfected miR-15a and pcDNA), miR-15a + pcDNA-SMAD2 group (co-transfected miR-15a mimics and pcDNA-SMAD2), were transfected into knee articular chondrocytes by liposome method, respectively. The cell proliferation and apoptosis of each group were detected by MTT assay and flow cytometry. The protein expression of SMAD2 was detected by Western blot. The fluorescence activity of each group was detected by dual luciferase reporter gene assay. Results: The expression of miR-15a in knee arthritis chondrocytes was significantly increased (p < .05) compared with that in normal chondrocytes. Moreover, overexpression of miR-15a and silencing of SMAD2 inhibited proliferation and promoted apoptosis in knee arthritis chondrocyte. MiR-15a targeted SMAD2. Overexpression of SMAD2 reversed the inhibitory effects on proliferation and promotion effects on apoptosis induced by miR-15a in knee arthritis chondrocytes. Conclusion: miR-15a can inhibit the proliferation and promote apoptosis of knee arthritis chondrocytes. The mechanism may be related to SMAD2, which will provide a new target for the treatment of knee arthritis.


Asunto(s)
Apoptosis/genética , Condrocitos/citología , Articulación de la Rodilla/citología , MicroARNs/genética , Proteína Smad2/genética , Artritis/genética , Artritis/patología , Secuencia de Bases , Proliferación Celular/genética , Condrocitos/patología , Regulación de la Expresión Génica/genética , Humanos , Articulación de la Rodilla/patología
3.
BMC Musculoskelet Disord ; 20(1): 316, 2019 Jul 06.
Artículo en Inglés | MEDLINE | ID: mdl-31279341

RESUMEN

BACKGROUND: Synovial mesenchymal stem cells (MSCs) are an attractive cell source for cartilage and meniscus regeneration. The optimum cryopreservation medium has not been determined, but dimethylsulfoxide (DMSO) should be excluded, if possible, because of its toxicity. The purposes of our study were to examine the possible benefits of higher concentrations of serum and the effectiveness of 100% serum (without DMSO) for the cryopreservation of synovial MSCs. METHODS: Human synovium was harvested from the knees of four donors with osteoarthritis during total knee arthroplasty. Synovial MSCs (8 × 105 cells) were suspended in 400 µL medium and used as a Time 0 control. The same number of synovial MSCs was also suspended in 400 µL α-MEM medium containing 10% fetal bovine serum (FBS) (5% DMSO, and 1% antibiotic), 95% FBS (and 5% DMSO), or 100% FBS (no DMSO) and cryopreserved at - 80 °C for 7 days. After thawing, the cell suspensions (1.5 µL; 3 × 103 cells) were cultured in 60 cm2 dishes for 14 days for colony formation assays. Additional 62.5 µL samples of cell suspensions (1.25 × 105 cells) were added to tubes and cultured for 21 days for chondrogenesis assays. RESULTS: Colony numbers were significantly higher in the Time 0 and 95% FBS groups than in the 10% FBS group (n = 24). Colony numbers were much lower in the 100% FBS group than in the other three groups. The cell numbers per dish reflected the colony numbers. Cartilage pellet weights were significantly heavier in the 95% FBS group than in the 10% FBS group, whereas no difference was observed between the Time 0 and the 95% FBS groups (n = 24). No cartilage pellets formed at all in the 100% FBS group. CONCLUSION: Synovial MSCs cryopreserved in 95% FBS with 5% DMSO maintained their colony formation and chondrogenic abilities to the same levels as observed in the cells before cryopreservation. Synovial MSCs cryopreserved in 100% FBS lost their colony formation and chondrogenic abilities.


Asunto(s)
Condrogénesis/efectos de los fármacos , Criopreservación/métodos , Crioprotectores/farmacología , Células Madre Mesenquimatosas , Membrana Sinovial/citología , Anciano , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Crioprotectores/química , Dimetilsulfóxido/química , Dimetilsulfóxido/farmacología , Femenino , Humanos , Articulación de la Rodilla/citología , Trasplante de Células Madre Mesenquimatosas , Osteoartritis/terapia , Suero/química
4.
BMC Musculoskelet Disord ; 20(1): 232, 2019 May 18.
Artículo en Inglés | MEDLINE | ID: mdl-31103042

RESUMEN

BACKGROUND: In osteoarthritis (OA), cartilage matrix is lost despite vigorous chondrocyte anabolism. In this study, we attempted to determine whether altered matrix synthesis is involved in this paradox in disease progression through gene expression analysis and ultrastructural analysis of collagen fibrils within the cartilage matrix. METHODS: Cartilage tissues were obtained from 29 end-stage OA knees and 11 control knees. First, cDNA microarray analysis was performed and the expression of 9 genes involved in collagen fibrillogenesis was compared between OA and control cartilages. Then their expression was investigated in further detail by a quantitative polymerase chain reaction (qPCR) analysis combined with laser capture microdissection. Finally, collagen fibril formation was compared between OA and control cartilage by transmission electron microscopy. RESULTS: The result of the microarray analysis suggested that the expression of type IX and type XI collagens and fibrillogenesis-related small leucine-rich proteoglycans (SLRPs) may be reduced in OA cartilage relative to the type II collagen expression. The qPCR analysis confirmed these results and further indicated that the relative reduction in the minor collagen and SLRP expression may be more obvious in degenerated areas of OA cartilage. An ultrastructural analysis suggested that thicker collagen fibrils may be formed by OA chondrocytes possibly through reduction in the minor collagen and SLRP expression. CONCLUSIONS: This may be the first study to report the possibility of altered collagen fibrillogenesis in OA cartilage. Disturbance in collagen fibril formation may be a previously unidentified mechanism underlying the loss of cartilage matrix in OA.


Asunto(s)
Cartílago Articular/patología , Colágeno Tipo IX/metabolismo , Colágeno Tipo XI/metabolismo , Osteoartritis de la Rodilla/patología , Proteoglicanos Pequeños Ricos en Leucina/metabolismo , Anciano , Anciano de 80 o más Años , Cartílago Articular/citología , Cartílago Articular/ultraestructura , Colágeno Tipo IX/ultraestructura , Colágeno Tipo XI/ultraestructura , Matriz Extracelular/patología , Matriz Extracelular/ultraestructura , Perfilación de la Expresión Génica , Humanos , Articulación de la Rodilla/citología , Articulación de la Rodilla/patología , Captura por Microdisección con Láser , Microscopía Electrónica de Transmisión
5.
FEBS Open Bio ; 9(6): 1144-1152, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-31037830

RESUMEN

Osteoarthritis (OA) is a chronic degenerative disease that commonly affects the elderly. Current drug therapies for treating OA may cause adverse side effects, and so there remains a need to develop alternative treatments. Bergapten (BG) is a coumarin phytohormone that is widely found in fruits and has antioxidative and anti-inflammatory effects. Here, we tested the hypothesis that BG may restrict the progression of OA by examining its effect on OA chondrocytes. We observed that BG significantly ameliorated interleukin (IL)-1ß-induced expression of inflammatory cytokines and mediators, including interleukin 1 (Il-1), interleukin 6 (Il-6), tumor necrosis factor α (Tnf-α), cyclooxygenase 2 (Cox-2) and matrix metalloproteinase 13 (Mmp-13), maintained chondrocyte phenotype, and promoted the secretion of cartilage-specific extracellular matrix. We provide evidence that BG exerts its anti-inflammatory effect by activating the ANP32A/ATM signaling pathway, which was recently verified to be associated with OA. In conclusion, these findings indicate that BG may be a potential candidate for treatment of OA.


Asunto(s)
5-Metoxipsoraleno/farmacología , Antiinflamatorios/farmacología , Proteínas de la Ataxia Telangiectasia Mutada/metabolismo , Proteínas Nucleares/metabolismo , Osteoartritis/metabolismo , Extractos Vegetales/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Condrocitos/efectos de los fármacos , Ciclooxigenasa 2/metabolismo , Citocinas/metabolismo , Ficus/química , Inflamación/tratamiento farmacológico , Articulación de la Rodilla/citología , Metaloproteinasa 13 de la Matriz/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos
6.
Artif Cells Nanomed Biotechnol ; 47(1): 1321-1325, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31007061

RESUMEN

In the process of natural precursors examining for the carbon dots (CDs) synthesis, a bio-friendly and highly luminescent CDs synthesis is being reported herein. For the first time, we are reporting CDs synthesis using Selenicereus grandiflorus plant materials without any use of additional oxidizing agents like ethanol which was used in the earlier mentioned reports. Ultraviolet-visible spectroscopy (UV-Vis), X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, high-resolution transmission electron (HR-TEM) microscopy and Raman spectroscopy. This method is entirely safe to use in all biological practices because no toxic chemicals were used in this method. Further, we studied the toxicity of CDs against chondrocytes obtained from the knee joint. In order to inquire about the feasible harmful effects of exposing CD on the knee cells, we carried out CD treatment in SW-1353 chondrocytes. Cytotoxicity outcomes displayed a dosage-dependent reduction of cell viability. Therefore, we studied the toxic effects of CDs on the knee, indicating the important prospectives of CDs in future knee therapy.


Asunto(s)
Carbono/química , Carbono/farmacología , Condrocitos/efectos de los fármacos , Articulación de la Rodilla/citología , Sustancias Luminiscentes/química , Sustancias Luminiscentes/farmacología , Puntos Cuánticos/química , Animales , Carbono/uso terapéutico , Supervivencia Celular/efectos de los fármacos , Técnicas de Química Sintética , Articulación de la Rodilla/efectos de los fármacos , Sustancias Luminiscentes/síntesis química , Sustancias Luminiscentes/uso terapéutico , Masculino , Ratas
7.
Rheumatology (Oxford) ; 58(5): 897-907, 2019 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-30085131

RESUMEN

OBJECTIVE: In this work, we aimed to elucidate the molecular mechanisms driving primary OA. By studying the dynamics of protein expression in two different types of OA joints we searched for similarities and disparities to identify key molecular mechanisms driving OA. METHODS: For this purpose, human SF samples were obtained from CMC-I OA and knee joint of OA patients. SF samples were analysed by label-free quantitative liquid chromatography mass spectrometry. Disease-relevant proteins identified in proteomics studies, such as clusterin, paraoxonase/arylesterase 1 (PON1) and transthyretin were validated by enzyme-linked immunosorbent assays, and on the mRNA level by droplet digital PCR. Functional studies were performed in vitro using primary chondrocytes. RESULTS: Differential proteomic changes were observed in the concentration of 40 proteins including clusterin, PON1 and transthyretin. Immunoassay analyses of clusterin, PON1, transthyretin and other inflammatory cytokines confirmed significant differences in protein concentration in SF of CMC-I and knee OA patients, with primarily lower protein expression levels in CMC-I. Functional studies on chondrocytes unequivocally demonstrated that stimulation with SF obtained from knee OA, in contrast to CMC-I OA joint, caused a significant upregulation in pro-inflammatory response, cell death and hypertrophy. CONCLUSION: This study demonstrates that differential expression of molecular players in SF from different OA joints evokes diverse effects on primary chondrocytes. The pathomolecular mechanisms of OA may significantly differ in various joints, a finding that brings a new dimension into the pathogenesis of primary OA.


Asunto(s)
Articulaciones Carpometacarpianas/metabolismo , Articulación de la Rodilla/metabolismo , Osteoartritis de la Rodilla/metabolismo , Líquido Sinovial/metabolismo , Articulaciones Carpometacarpianas/citología , Condrocitos/metabolismo , Ensayo de Inmunoadsorción Enzimática , Humanos , Articulación de la Rodilla/citología , Espectrometría de Masas , Proteómica , ARN Mensajero/metabolismo
8.
Cartilage ; 10(1): 36-42, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-29322876

RESUMEN

DESIGN: In the process of cell division, the extremes of the eukaryotic chromosomes are progressively shortening, and this phenomenon is related to cell degeneration and senescence. The treatment of cartilage lesions with autologous chondrocytes implies that cells proliferate in an artificial environment. We have studied the viability of cultured chondrocytes after measurement of their telomere length before implantation. METHODS: Articular cartilage biopsies (B1, B2, and B3) were obtained from 3 patients (2 males and 1 female) with knee cartilage defects, who were going to be treated with chondrocyte implantation. Chondrocytes were cultured in DMEM with autologous serum. After the third passage, an aliquot of 1 million cells was removed to estimate the telomere length and the remaining cells were implanted. Telomere length was measured by quantitative fluorescent in situ hybridization (Q-FISH). Patients' clinical outcome was determined preoperatively, and 12 and 24 months postimplantation with the International Knee Documentation Committee (IKDC) questionnaire. RESULTS: After chondrocyte implantation, IKDC score doubled at 12 and 24 months with regard to the basal value. After 3 passages, chondrocytes were cultured for a mean of 45.67 days, the mean duplication time being 4.53 days and the mean number of cell divisions being 10.04 during the culture period. The 20th percentile of telomere lengths were 6.84, 6.96, and 7.06 kbp and the median telomere lengths 10.30, 10.47, and 10.73 kbp, respectively. No significant correlation was found between IKDC score and telomere length. CONCLUSION: Culturing autologous chondrocytes for implantation is not related to cell senescence in terms of telomere length.


Asunto(s)
Enfermedades de los Cartílagos/patología , Cartílago Articular/citología , Condrocitos/patología , Trasplante de Células Madre , Telómero/patología , Adulto , Enfermedades de los Cartílagos/terapia , Cartílago Articular/patología , Células Cultivadas , Femenino , Humanos , Hibridación Fluorescente in Situ , Articulación de la Rodilla/citología , Articulación de la Rodilla/patología , Masculino , Trasplante Autólogo
9.
Cartilage ; 10(1): 61-69, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-28486813

RESUMEN

OBJECTIVE: The aim of this study was to evaluate an intraarticular injection of different doses of autologous mesenchymal stem cells (MSCs) for improving repair of midterm osteochondral defect. DESIGN: At 4 weeks postoperative marrow stimulation model bilaterally (3 mm diameter; 4 mm depth) in the medial femoral condyle, autologous MSCs were injected into knee joint. Twenty-four Japanese rabbits aged 6 months were divided randomly into 4 groups ( n = 6 per group): the control group and and MSC groups including 0.125, 1.25, and 6.25 million MSCs. Repaired tissue was assessed macroscopically and histologically at 4 and 12 weeks after intraarticular injection of MSCs. RESULTS: At 12 weeks, there was no repair tissue in the control group. The gross appearance of the 1.25 and 6.25 million MSC groups revealed complete repair of the defect with white to pink tissue at 12 weeks. An osteochondral repair was histologically significantly better in the 1.25 and 6.25 million MSC groups than in the control and 0.125 million MSC groups at 4 and 12 weeks, due to presence of hyaline-like tissue in the deep layer at 4 weeks, and at 12 weeks hyaline cartilage formation at the periphery and fibrous tissue containing some chondrocytes in the deep layer of the center of the defect. Subchondral bone was restructured in the 1.25 and 6.25 million MSC groups, although it did not resemble the normal bone. CONCLUSION: An intraarticular injection of 1.25 or 6.25 million MSCs could promote the repair of subchondral bone, even in the case of midterm osteochondral defect.


Asunto(s)
Cartílago Articular/citología , Condrocitos/fisiología , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/fisiología , Osteocondritis/terapia , Animales , Recuento de Células , Condrogénesis , Inyecciones Intraarticulares , Articulación de la Rodilla/citología , Articulación de la Rodilla/patología , Osteocondritis/patología , Osteocondritis/fisiopatología , Conejos , Distribución Aleatoria , Trasplante Autólogo
10.
Cartilage ; 10(1): 111-119, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-28715962

RESUMEN

OBJECTIVE: In the present in vitro study, we analyzed the chondrogenic differentiation capacity of human chondrocytes postmortally isolated from unaffected knee cartilage by the addition of transforming growth factor-ß1 (TGF-ß1) and/or insulin-like growth factor-1 (IGF-1) and different oxygen levels. DESIGN: After 14 and 35 days, DNA concentrations and protein contents of Col1, Col2, aggrecan as well as glycosaminoglycans (GAGs) of chondrocytes cultivated as pellet cultures were analyzed. Additionally, expression rates of mesenchymal stem cell (MSC)-associated differentiation markers were assessed in monolayer cultures. RESULTS: All cultivated chondrocytes were found to be CD29+/CD44+/CD105+/CD166+. Chondrocytic pellets stimulated with TGF-ß1 showed enhanced synthesis rates of hyaline cartilage markers and reduced expression of the non-hyaline cartilage marker Col1 under hypoxic culture conditions. CONCLUSIONS: Our results underline the substantial chondrogenic potential of human chondrocytes postmortally isolated from unaffected articular knee cartilage especially in case of TGF-ß1 administration.


Asunto(s)
Diferenciación Celular/fisiología , Condrocitos/fisiología , Factor I del Crecimiento Similar a la Insulina/metabolismo , Oxígeno/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Agrecanos/metabolismo , Cartílago Articular/citología , Técnicas de Cultivo de Célula , Células Cultivadas , Condrogénesis , Glicosaminoglicanos/metabolismo , Humanos , Articulación de la Rodilla/citología , Células Madre Mesenquimatosas
11.
J Biomech ; 83: 65-75, 2019 01 23.
Artículo en Inglés | MEDLINE | ID: mdl-30501912

RESUMEN

Site-specific and depth-dependent properties of cartilage were implemented within a finite element (FE) model to determine if compositional or structural changes in the tissue could explain site-specific alterations of chondrocyte deformations due to cartilage loading in rabbit knee joints 3 days after a partial meniscectomy (PM). Depth-dependent proteoglycan (PG) content, collagen content and collagen orientation in the cartilage extracellular matrix (ECM), and PG content in the pericellular matrix (PCM) were assessed with microscopic and spectroscopic methods. Patellar, femoral groove and samples from both the lateral and medial compartments of the femoral condyle and tibial plateau were extracted from healthy controls and from the partial meniscectomy group. For both groups and each knee joint site, axisymmetric FE models with measured properties were generated. Experimental cartilage loading was applied in the simulations and chondrocyte volumes were compared to the experimental values. ECM and PCM PG loss occurred within the superficial cartilage layer in the PM group at all locations, except in the lateral tibial plateau. Collagen content and orientation were not significantly altered due to the PM. The FE simulations predicted similar chondrocyte volume changes and group differences as obtained experimentally. Loss of PCM fixed charge density (FCD) decreased cell volume loss, as observed in the medial femur and medial tibia, whereas loss of ECM FCD increased cell volume loss, as seen in the patella, femoral groove and lateral femur. The model outcome, cell volume change, was also sensitive to applied tissue geometry, collagen fibril orientation and loading conditions.


Asunto(s)
Cartílago Articular/citología , Condrocitos/citología , Análisis de Elementos Finitos , Articulación de la Rodilla/citología , Articulación de la Rodilla/cirugía , Fenómenos Mecánicos , Meniscectomía , Animales , Tamaño de la Célula , Colágeno/metabolismo , Matriz Extracelular/metabolismo , Proteoglicanos/metabolismo , Conejos
12.
Theranostics ; 8(20): 5519-5528, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30555561

RESUMEN

Rationale: Recent studies confirmed that osteoarthritis (OA) is associated with systemic inflammation. Adipose-derived stromal cells (ASCs) could become the most promising cell-based therapy in OA, based not only on their differentiation capacities and trophic and paracrine effects on the existing cartilage, but also on their immunomodulatory properties. Here, we wanted to determine the biological effect of autologous ASC intra-articular (IA) injection. Method: To this aim, we monitored the profile of immune cells in fresh peripheral blood after IA injection of autologous ASCs in the knee of 18 patients with severe OA (ADIPOA phase I study). Specifically, we used 8-color flow cytometry antibody panels to characterize the frequencies of innate and adaptive immune cell subsets (monocytes, dendritic cells, regulatory T cells and B cells) in blood samples at baseline (before injection) and one week, one month and three months after ASC injection. Results: We found that the percentage of CD4+CD25highCD127lowFOXP3+ regulatory T cells was significantly increased at 1 month after ASC injection, and this effect persisted for at least 3 months. Moreover, CD24highCD38high transitional B cells also were increased, whereas the percentage of classical CD14+ monocytes was decreased, at 3 months after ASC injection. These results suggest a global switch toward regulatory immune cells following IA injection of ASCs, underscoring the safety of ASC-based therapy. We did not find any correlation between the scores for the Visual Analogic Scale for pain, the Western Ontario and McMaster Universities Osteoarthritis Index (pain subscale and total score) at baseline and the immune cell profile changes, but this could be due to the small number of analyzed patients. Conclusion: ASCs may drive an immediate local response by releasing paracrine factors and cytokines, and our results suggest that ASCs could also initiate a cascade resulting in a long-lasting systemic immune modulation.


Asunto(s)
Tejido Adiposo/citología , Células Madre Mesenquimatosas/citología , Osteoartritis de la Rodilla/inmunología , Osteoartritis de la Rodilla/terapia , Células del Estroma/citología , Citometría de Flujo , Humanos , Articulación de la Rodilla/citología , Articulación de la Rodilla/inmunología , Estudios Prospectivos
13.
Acta Chir Orthop Traumatol Cech ; 85(5): 366-369, 2018.
Artículo en Checo | MEDLINE | ID: mdl-30383534

RESUMEN

The authors present an overview of the commonly used techniques and new trends of the cartilage imaging, especially postoperatively, and also discuss the potential of MRI imaging of the cartilage from the perspective of an experienced orthopaedic surgeon. In conclusion, the authors propose possible explanations for the potential discrepancies between the MRI and the arthroscopic findings. Hyaline cartilage damage and subsequent reparation of this tissue is one of the topical issues of orthopaedics and traumatology. Due to the expanding possibilities of treatment of this tissue and a relatively good effect of the surgery, the number of patients indicated for magnetic resonance imaging prior to the surgery has been on an increase. To make a decision concerning the subsequent type of treatment, it is necessary to get an idea of the cartilage cover condition, articular surfaces and also of the associated pathologies. The degree of cartilage damage can be assessed by arthroscopy or magnetic resonance imaging, which provides also the possibility of the subchondral lesion detection. Thanks to the noninvasive nature of the MRI examination, it has become the most important method in full imaging of the articular cartilage. The MRI of the cartilage has many options and at present the evaluation of the hyaline cartilage should be an integral part of each MRI examination of joints. For a more accurate assessment of the cartilage there are several advanced techniques available that can be used not only for preoperative diagnostics, but also for monitoring after the surgery. Key words: hyaline cartilage, magnetic resonance, arthroscopy.


Asunto(s)
Cartílago Articular/diagnóstico por imagen , Cartílago Hialino/diagnóstico por imagen , Articulación de la Rodilla/citología , Artroscopía/métodos , Cartílago Articular/patología , Toma de Decisiones/ética , Humanos , Cartílago Hialino/patología , Articulación de la Rodilla/cirugía , Imagen por Resonancia Magnética/métodos , Imagen por Resonancia Magnética/normas , Cirujanos Ortopédicos , Periodo Posoperatorio , Cuidados Preoperatorios/normas , Radiólogos
14.
ACS Appl Mater Interfaces ; 10(45): 38715-38728, 2018 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-30360061

RESUMEN

In situ tissue regeneration by homing endogenous reparative cells to the injury site has been extensively researched as a promising alternative strategy to facilitate tissue repair. In this study, a promising scaffolding system DCM-RAD/SKP, which integrated a decellularized cartilage matrix (DCM)-derived scaffold with a functionalized self-assembly Ac-(RADA)4-CONH2/Ac-(RADA)4GGSKPPGTSS-CONH2 (RAD/SKP) peptide nanofiber hydrogel, was designed for repairing rabbit osteochondral defect. In vitro experiments showed that rabbit bone marrow stem cells migrated into and have higher affinity toward the functional scaffolding system DCM-RAD/SKP than the control scaffolds. One week after in vivo implantation, the functional scaffolding system DCM-RAD/SKP facilitated the recruitment of endogenous mesenchymal stem cells within the defect site. Moreover, gene expression analysis indicated that the DCM-RAD/SKP promoted chondrogenesis of the recruited cells. In vivo results showed that the DCM-RAD/SKP achieved superior hyaline-like cartilage repair and successful subchondral bone reconstruction. By contrast, the control groups mostly led to fibrous tissue repair. These findings indicate that the DCM-RAD/SKP can recruit endogenous stem cells into the site of cartilage injury and promote differentiation of the infiltrating cells into the chondrogenic lineage, holding great potential as a one-step surgery strategy for cartilage repair.


Asunto(s)
Células de la Médula Ósea/citología , Cartílago Articular/fisiología , Hidrogeles/administración & dosificación , Células Madre Mesenquimatosas/citología , Oligopéptidos/administración & dosificación , Regeneración/fisiología , Andamios del Tejido , Animales , Cartílago Articular/citología , Articulación de la Rodilla/citología , Articulación de la Rodilla/cirugía , Conejos , Porcinos , Microtomografía por Rayos X
15.
PLoS One ; 13(8): e0201839, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30071108

RESUMEN

Sclerostin is a potent inhibitor of the canonical Wnt signaling pathway. Wnt signaling pathways have multiple roles in the regulation of cartilage development, growth, and maintenance. This study focused on the role of sclerostin in the process of chondrogenic differentiation. We hypothesized that sclerostin is essential to induce chondrogenic differentiation and regulate endochondral ossification. ATDC5 cells were used to investigate chondrogenic differentiation and terminal calcification. During chondrogenic differentiation, intrinsic sclerostin was upregulated in the early stage, but downregulated in the late stage. Addition of sclerostin elevated expressions of Sox9 and Col2a1 (P<0.05) and reduced expressions of Runx2, Col10a1, MMP-3, MMP-13, and ADAMTS5 (P<0.05) through inhibition of the Wnt-ß-catenin signaling pathway (P<0.05). Terminal calcification was significantly inhibited by sclerostin (P<0.05). In contrast, deletion of sclerostin decreased expressions of Sox9 and Col2a1 (P<0.05), increased expressions of Runx2, Col10a1, MMP-3, and MMP-13 (P<0.05), and promoted terminal calcification (P<0.05). This study provides insights into the possible role of sclerostin in the regulation of chondrogenic differentiation. Sclerostin is upregulated in the early stage of chondrogenic differentiation, but is not required in endochondral ossification. Sclerostin is a candidate modulator for chondrogenic differentiation.


Asunto(s)
Condrocitos/metabolismo , Condrogénesis/fisiología , Glicoproteínas/metabolismo , Articulación de la Rodilla/crecimiento & desarrollo , Articulación de la Rodilla/metabolismo , Osteogénesis/fisiología , Animales , Diferenciación Celular/fisiología , Línea Celular Tumoral , Condrocitos/citología , Silenciador del Gen , Glicoproteínas/genética , Péptidos y Proteínas de Señalización Intercelular , Articulación de la Rodilla/citología , Ratones Endogámicos C57BL , ARN Interferente Pequeño , Regulación hacia Arriba
16.
J Biomech ; 77: 91-98, 2018 08 22.
Artículo en Inglés | MEDLINE | ID: mdl-30049448

RESUMEN

In order to pre-clinically evaluate the performance and efficacy of novel osteochondral interventions, physiological and clinically relevant whole joint simulation models, capable of reproducing the complex loading and motions experienced in the natural knee environment are required. The aim of this study was to develop a method for the assessment of tribological performance of osteochondral grafts within an in vitro whole natural joint simulation model. The study assessed the effects of osteochondral allograft implantation (existing surgical intervention for the repair of osteochondral defects) on the wear, deformation and damage of the opposing articular surfaces. Tribological performance of osteochondral grafts was compared to the natural joint (negative control), an injury model (focal cartilage defects) and stainless steel pins (positive controls). A recently developed method using an optical profiler (Alicona Infinite Focus G5, Alicona Imaging GmbH, Austria) was used to quantify and characterise the wear, deformation and damage occurring on the opposing articular surfaces. Allografts inserted flush with the cartilage surface had the lowest levels of wear, deformation and damage following the 2 h test; increased levels of wear, deformation and damage were observed when allografts and stainless steel pins were inserted proud of the articular surface. The method developed will be applied in future studies to assess the tribological performance of novel early stage osteochondral interventions prior to in vivo studies, investigate variation in surgical precision and aid in the development of stratified interventions for the patient population.


Asunto(s)
Aloinjertos , Articulación de la Rodilla/citología , Fenómenos Mecánicos , Modelos Biológicos , Animales , Fenómenos Biomecánicos , Humanos , Porcinos , Trasplante Homólogo
17.
Biomed Eng Online ; 17(1): 42, 2018 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-29665801

RESUMEN

BACKGROUND: OpenSim musculoskeletal models provide an accurate simulation environment that eases limitations of in vivo and in vitro studies. In this work, a biomechanical knee model was formulated with femoral articular cartilages and menisci along with 25 connective tissue bundles representing ligaments and capsules. The strain patterns of the connective tissues in the presence of femoral articular cartilage and menisci in the OpenSim knee model was probed in a first of its kind study. METHODS: The effect of knee flexion (0°-120°), knee rotation (- 40° to 30°) and knee adduction (- 15° to 15°) on the anterior cruciate, posterior cruciate, medial collateral, lateral collateral ligaments and other connective tissues were studied by passive simulation. Further, a new parameter for assessment of strain namely, the differential inter-bundle strain of the connective tissues were analyzed to provide new insights for injury kinematics. RESULTS: ACL, PCL, LCL and PL was observed to follow a parabolic strain pattern during flexion while MCL represented linear strain patterns. All connective tissues showed non-symmetric parabolic strain variation during rotation. During adduction, the strain variation was linear for the knee bundles except for FL, PFL and TL. CONCLUSIONS: Strains higher than 0.1 were observed in most of the bundles during lateral rotation followed by abduction, medial rotation and adduction. In the case of flexion, highest strains were observed in aACL and aPCL. A combination of strains at a flexion of 0° with medial rotation of 30° or a flexion of 80° with rotation of 30° are evaluated as rupture-prone kinematics.


Asunto(s)
Tejido Conectivo , Articulación de la Rodilla/citología , Modelos Biológicos , Estrés Mecánico , Fenómenos Biomecánicos , Femenino , Humanos , Articulación de la Rodilla/fisiología , Rango del Movimiento Articular
18.
Exp Anim ; 67(3): 349-359, 2018 Jul 30.
Artículo en Inglés | MEDLINE | ID: mdl-29515059

RESUMEN

Rabbit mesenchymal stem cells (MSCs) are important seed cells in regenerative medicine research, particularly in translational research. In the current study, we showed that rabbit subchondral bone is a reliable source of MSCs. First, we harvested subchondral bone (SCB) from the rabbit knee-joint and initiated the MSC culture by cultivating enzyme-treated SCB. Adherent fibroblast-like cells that outgrew from SCB fulfill the common immuno-phenotypic criteria for defining MSCs, but with low contamination of CD45+ hematopoietic cells. Interestingly, differentiated SCB-MSCs expressed osteogenic and chondrogenic markers at significantly higher levels than those in bone marrow cell suspension-derived MSCs (BMS-MSCs) (P<0.05). No differences in the expression of adipogenic markers between SCB-MSC and BMS-MSC (P>0.05) were observed. Moreover, the results of the colony forming unit-fibroblast assay and sphere formation assay demonstrated that the SCB-MSCs had increased self-renewal potential. SCB-MSCs expressed higher levels of the stemness markers Nanog, OCT4, and Sox-2 compared to in BMS-MSCs (P<0.05). Furthermore, the results of both the CCK-8-based assay and CFSE dilution assay showed that SCB-MSCs exhibited enhanced proliferative capacity. In addition, SCB-MSCs exhibited higher phosphorylation of extracellular signal-related kinase/mitogen-activated protein kinase signaling, which is closely related to MSC proliferation. In conclusion, we identified SCB-MSCs as a novel stem cell population that met the requirements of MSCs; the unique properties of SCB-MSC are important for the potential treatment of tissue damage resulting from disease and trauma.


Asunto(s)
Huesos/citología , Cartílago/citología , Diferenciación Celular , Proliferación Celular , Autorrenovación de las Células , Condrogénesis , Articulación de la Rodilla/citología , Células Madre Mesenquimatosas/citología , Osteogénesis , Animales , Técnicas de Cultivo de Célula , Células Cultivadas , Masculino , Conejos
19.
Cell Tissue Bank ; 19(3): 399-404, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29453700

RESUMEN

The purpose of this study is to evaluate the reliability of cartilage digestion and fluorescein diacetate-ethidium bromide (FDA-EB) fluorescence staining for the detection of chondrocyte viability in osteochondral grafts. Sixteen fresh osteochondral grafts were harvested from pig knee condyles, and the articular cartilage tissue was preserved. Each cartilage graft was cut into two 70-µm thick pieces and randomly allocated to Group A or Group B. The cell viability of Group A was detected using FDA-EB fluorescence staining of the digested cartilage, and the viability of Group B was detected with FDA-EB fluorescence staining of cartilage sections. Comparisons of chondrocyte viability and correlation analyses of the two groups were performed using the paired sample t test and Pearson correlation test, respectively. No significant difference was found in the chondrocyte viability between Groups A and B (p > 0.05), and a strong correlation was observed (r = 0.70, p < 0.05). Therefore, cartilage digestion with FDA-EB fluorescence staining is a reliable method for detecting chondrocyte viability in osteochondral grafts.


Asunto(s)
Cartílago Articular/citología , Condrocitos/citología , Etidio/análisis , Fluoresceínas/análisis , Colorantes Fluorescentes/análisis , Animales , Supervivencia Celular , Fluorescencia , Articulación de la Rodilla/citología , Masculino , Microtomía/métodos , Coloración y Etiquetado/métodos , Porcinos
20.
Int Immunopharmacol ; 55: 282-289, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29291543

RESUMEN

OBJECTIVE: Platelet-rich plasma (PRP) has been reported to alleviate degenerative pathological damage to joint cartilage. This study aimed to investigate the effect of PRP on Wnt/ß-catenin signaling in rabbit chondrocytes. METHODS: Using 3-month-old New Zealand white rabbits, PRP was prepared from venous blood, and chondrocytes were cultured from knee joint cartilage and identified by staining for type II collagen and proteoglycan. The effects of PRP on chondrocyte viability were measured. The chondrocytes were divided into 5 groups: control, IL-1ß, PRP (100-fold dilution), Dkk-1 (100ng/mL) and Dkk-1+PRP. The IL-1ß, PRP, Dkk-1 and Dkk-1+PRP groups were treated with interleukin (IL)-1ß (50µL, 10µg/mL) for24h. Chondrocyte morphology was observed by electron microscopy. Levels of carboxy terminal peptide (CTX-II) and cartilage oligomeric matrix protein (COMP) in culture media were measured by ELISA. Wnt-1, ß-catenin and GSK-3ß mRNA and protein expression were determined by RT-PCR and western blot respectively. RESULTS: PRP enhanced chondrocyte proliferation. Chondrocytes in the IL-1ß group showed ultrastructural abnormalities that were less pronounced in the PRP, Dkk-1 and Dkk-1+PRP groups. CTX-II and COMP concentrations were higher in the IL-1ß group than in the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). The IL-1ß group had higher mRNA and protein Wnt1 and ß-catenin levels and lower GSK-3ß levels than the control, PRP, Dkk-1 and Dkk-1+PRP groups (P<0.05). CONCLUSION: PRP may protect chondrocytes activated by IL-1ß via inhibiting Wnt/ß-catenin signaling.


Asunto(s)
Condrocitos/fisiología , Articulación de la Rodilla/citología , Plasma Rico en Plaquetas/metabolismo , Animales , Proteína de la Matriz Oligomérica del Cartílago/metabolismo , Supervivencia Celular , Células Cultivadas , Colágeno Tipo II/metabolismo , Glucógeno Sintasa Quinasa 3 beta/genética , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Interleucina-1beta/inmunología , Osteoartritis , Conejos , Vía de Señalización Wnt , Proteína Wnt1/genética , Proteína Wnt1/metabolismo , beta Catenina/genética , beta Catenina/metabolismo
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