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Rev. bras. ortop ; 56(4): 470-477, July-Aug. 2021. tab, graf
Article in English | LILACS | ID: biblio-1341162


Abstract Objective The aim of our study is to analyze the clinical and functional results obtained using autologous chondrocytes embedded in a fibrin scaffold in knee joint injuries. Methods We included 56 patients, 36 men and 20 women, with a mean age 36 years. Six of the patients were professional athletes, with single knee injuries that were either chondral or osteochondral (43 chondral, 9 osteochondral, 2 cases of osteochondritis dissecans and 2 osteochondral fractures), 2 to 10 cm2 in size and ≤ 10 mm deep, with no signs of osteoarthritis. The location of the injury was in the patella (8), the medial femoral condyle (40) and lateral femoral condyle (7) and one in the trochlea. The mean follow-up was 3 (range: 1-6) years. The clinical course was assessed using the Cincinnati and Knee Injury and Osteoarthritis Outcome (KOOS) scores, 6 and 12 months after surgery. The paired Student t-test was used to compare pre-and postoperative results. Results Six months after the implant, patients resumed their everyday activities. On the assessment scores, their condition was improving in comparison with their presurgical state (p < 0.05). They were also able to carry out their sporting activities more easily than prior to surgery (p < 0.05). Conclusion The seeding of chondrocytes in fibrin may provide a favorable microenvironment for the synthesis of extracellular matrix and improved the clinical condition and activity of the patients 1 year after surgery.

Resumo Objetivo O objetivo do nosso estudo é analisar os resultados clínicos e funcionais do tratamento de lesões nas articulações do joelho com condrócitos autólogos embebidos em arcabouço de fibrina. Métodos O estudo foi realizado com 56 pacientes (36 homens e 20 mulheres) com idade média de 36 anos; 6 indivíduos eram atletas profissionais. Os pacientes apresentavam lesões únicas, condrais ou osteocondrais (43 condrais, nove osteocondrais, 2 casos de osteocondrite dissecante e duas fraturas osteocondrais) no joelho, com 2 a 10 cm2 de tamanho e ≤ 10 mm de profundidade, sem sinais de osteoartrite. As lesões estavam localizadas na patela (8), no côndilo femoral medial (40), no côndilo femoral lateral (7) e na tróclea (1). O período médio de acompanhamento foi de 3 anos (faixa de 1-6 anos). A evolução clínica foi avaliada pelos escores de Cincinnati e Knee Injury and Osteoarthritis Outcome (KOOS), 6 e 12 meses após a cirurgia. O teste t de Student pareado foi utilizado para comparação dos achados pré e pós-operatórios. Resultados Os pacientes retomaram suas atividades diárias 6 meses após o implante. Os escores avaliados demonstraram a melhora em comparação ao estado pré-cirúrgico (p < 0,05). Além disso, os pacientes conseguiram realizar suas atividades esportivas com mais facilidade do que antes da cirurgia (p < 0,05). Conclusão A cultura de condrócitos em fibrina pode proporcionar um microambiente favorável para a síntese de matriz extracelular e melhorar a condição clínica e a atividade dos pacientes 1 ano após a cirurgia

Humans , Male , Female , Fibrin , Cartilage , Chondrocytes , Scaphoid Bone , Knee
Article in Chinese | WPRIM | ID: wpr-921944


OBJECTIVE@#To investigate the effect of the exosomes from bone marrow mesenchymal stem cells (BMSCs) transfected with silence plasmid of Piezol small interference RNA (siRNA)on osteoarthritis (OA) animal model.@*METHODS@#Twenty male SD rats with specific pathogen free (SPF) were selected, ranging in age from 5.46 to 6.96 months, with a mean of (6.21± 0.75) months;and ranging in weight from 385.76 g to 428.66 g, with a mean of (407.21±21.45) g. BMSCs were extracted. The siRNA silencing plasmid of piezo1 was constructed by siRNA technology. After lentivirus was transfected into BMSCs, the exosomes were extracted. At the cellular level, BMSCs were divided into blank plasmid group and siRNA silencing plasmid group according to whether siRNA-Piezo1 was transfected or not. The osteogenic induction ability of siRNA-Piezo1 on BMSCs was detected by RT-PCR and Western blot. At the animal model level, the OA model was established by surgical resection of cruciate ligament of knee joint.According to different treatment schemes, SD rats were divided into 4 groups:blank control group, model group, BMSCs group and exosome group. SD rats in the blank control group were not treated. In the model group, the cruciate ligaments of rats were excised and OA animal model was established. In BMSCs group, BMSCs were injected into knee joint under CT guidance after OA model establishment, and the cell volume was 5×10@*RESULTS@#The lentivirus transfection efficiency was(92.11±4.22)%. RT-PCR showed that the relative expression of Piezo1 mRNA in blank plasmid group was 1.07±0.06, which was significantly different from that of 0.31±0.01 in siRNA silencing plasmid group (@*CONCLUSION@#Piezo1 siRNA silencing vector can promote the differentiation of BMSCs into chondrocytes and effectively inhibit the progression of OA, so as to delay the disease of OA.

Animals , Chondrocytes , Disease Models, Animal , Exosomes/genetics , Male , Mesenchymal Stem Cells , Osteoarthritis/therapy , RNA, Small Interfering/genetics , Rats , Rats, Sprague-Dawley , Tomography, X-Ray Computed
Article in Chinese | WPRIM | ID: wpr-921929


Osteoarthritis(OA) is one of the most common joint diseases. As Chinese society enters the age of aging, the incidence of OA has been soar year by year, and research on its pathogenesis has been continuously valued by researchers. Studies have found that inflammatory cytokines, mainly interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), were responsible for the construction of OA inflammatory networks. It was also found that the overexpression of proteases, mainly matrix metalloproteinases(MMPs) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), was the direct cause of OA cartilage deficiency. What's more, signaling pathways such as stromal cell derived factor-1 (SDF-1) and Wnt, chondrocytic senescence and the senescence-associated secretory phenotype (SASP), chondrocyte apoptosis and autophagy, and estrogen all play significant roles in OA pathogenesis. This paper extensively reviews the research literature relevant to the pathogenesis of OA in recent years, and systematically expounds the pathogenesis of OA from two aspects:molecular level and cell level. At the end of the paper, we discussed and predicted some potential directions in the future clinical diagnosis and treatment of OA.

Cartilage , Cartilage, Articular , Chondrocytes , Humans , Interleukin-1beta , Osteoarthritis/genetics , Signal Transduction , Tumor Necrosis Factor-alpha
Article in English | WPRIM | ID: wpr-921387


OBJECTIVES@#To study the effects of 17β-estradiol (E2) on the regulation of the proliferation of condylar chondrocytes and provide a preliminary discussion on the role of phosphorylate-mammalian target of rapamycin (p-mTOR) in this regulatory process.@*METHODS@#Condylar chondrocytes were isolated from 6-week-old female rats for primary culture. Drug treatment with different concentrations of E2 and/or rapamycin (RAPA) was carried out on second-generation cells. Cell Counting Kit 8 was used to measure the cell viability of condylar chondrocytes after culture for 24, 48, or 72 h, and reverse transcription-polymerase chain reaction (RT-PCR) was applied to detect the relative gene expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), collagen type Ⅱ (COLⅡ), autophagy-related gene 6 (Beclin-1), and autophagy-related gene 5 (ATG-5). Western blot was employed to determine the relative protein expression of ERα, ERβ, Beclin-1, lipid-modified light chain 3B (LC3-Ⅱ), and p-mTOR.@*RESULTS@#E2 could significantly promote the proliferation of chondrocytes cultured @*CONCLUSIONS@#At a concentration of 10

Animals , Autophagy , Cell Proliferation , Chondrocytes , Estradiol , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta , Female , Phosphorylation , Rats
Article in English | WPRIM | ID: wpr-878418


Cartilage stem cells (CSCs) are cells that self-proliferate, have surface antigen expression, and have multidirectional differentiation potential in the articular cartilage. CSCs, as an ideal source of stem cells, has a good application prospect in stem cell therapy. This article reviews the CSCs markers, cartilage differentiation signaling pathway, and clinical treatment of osteoarthritis.

Cartilage, Articular , Cell Differentiation , Chondrocytes , Humans , Osteoarthritis , Stem Cells
Chinese Medical Journal ; (24): 963-970, 2021.
Article in English | WPRIM | ID: wpr-878129


BACKGROUND@#Histone deacetylase 4 (HDAC4) regulates chondrocyte hypertrophy and bone formation. The aim of the present study was to explore the effects of HDAC4 on Interleukin 1 beta (IL-1β)-induced chondrocyte extracellular matrix degradation and whether it is regulated through the WNT family member 3A (WNT3A)/β-catenin signaling pathway.@*METHODS@#Primary chondrocytes (CC) and human chondrosarcoma cells (SW1353 cells) were treated with IL-1β and the level of HDAC4 was assayed using Western blotting. Then, HDAC4 expression in the SW1353 cells was silenced using small interfering RNA to detect the effect of HDAC4 knockdown on the levels of matrix metalloproteinase 3 (MMP3) and MMP13 induced by IL-1β. After transfection with HDAC4 plasmids, the overexpression efficiency was examined using Real-time quantitative polymerase chain reaction (qRT-PCR) and the levels of MMP3 and MMP13 were assayed using Western blotting. After incubation with IL-1β, the translocation of β-catenin into the nucleus was observed using immunofluorescence staining in SW1353 cells to investigate the activation of the WNT3A/β-catenin signaling pathway. Finally, treatment with WNT3A and transfection with glycogen synthase kinase 3 beta (GSK3β) plasmids were assessed for their effects on HDAC4 levels using Western blotting.@*RESULTS@#IL-1β downregulated HDAC4 levels in chondrocytes and SW1353 cells. Furthermore, HDAC4 knockdown increased the levels of MMP3 and MMP13, which contributed to the degradation of the extracellular matrix. Overexpression of HDAC4 inhibited IL-1β-induced increases in MMP3 and MMP13. IL-1β upregulated the levels of WNT3A, and WNT3A reduced HDAC4 levels in SW1353 cells. GSK-3β rescued IL-1β-induced downregulation of HDAC4 in SW1353 cells.@*CONCLUSION@#HDAC4 exerted an inhibitory effect on IL-1β-induced extracellular matrix degradation and was regulated partially by the WNT3A/β-catenin signaling pathway.

Cell Line, Tumor , Cells, Cultured , Chondrocytes/metabolism , Glycogen Synthase Kinase 3 beta/genetics , Histone Deacetylases/genetics , Humans , Interleukin-1beta/pharmacology , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 3 , Repressor Proteins , Wnt Signaling Pathway , Wnt3A Protein/genetics , beta Catenin/metabolism
Article in English | WPRIM | ID: wpr-880618


OBJECTIVES@#Chondrocyte apoptosis is an important process in the pathogenesis of osteoarthritis. Mangiferin exerts multiple pharmacological effects such as anti-inflammatory and anti-apoptosis. However, the role of mangiferin in chondrocyte apoptosis is not clear. In this study, we aimed to explore the role of mangiferin in IL-1β-induced chondrocyte apoptosis.@*METHODS@#ATDC5 cells were randomly divided into a control group, a IL-1β group, a MFN-L group, a MFN-M group, a MFN-H group and a MFN+LY294002 group. Cells in the control group were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN-L group, the MFN-M group and the MFN-H group were pretreated with 5, 10 and 20 μmol/L mangiferin for 1 h respectively, and then they were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN+LY294002 group were treated with LY294002 (25 μmol/L) for 1 h, then mangiferin (20 μmol/L) and IL-1β (10 ng/mL) for 1 h and 24 h, respectively. Cell viability was detected by CCK-8 assay and cell apoptosis was measured by flow cytometry. Colorimetric assay was conducted to measure the caspase-3 activity. The protein levels of Bcl-2, Bax, and phosphoinositide 3-kinase (PI3K)/Akt signaling pathway related proteins were detected by Western blotting.@*RESULTS@#Compared to the control group, cell viability was significantly decreased; cell apoptosis, caspase-3 activity and Bax protein expression were significantly increased; the protein levels of Bcl-2, p-PI3K, and p-Akt were significantly decreased in the IL-1β group (all @*CONCLUSIONS@#Mangiferin could attenuate IL-1β-induced apoptosis of the mice chondrocytes, which is mediated by the activation of PI3K/Akt signaling pathway.

Animals , Apoptosis , Chondrocytes , Interleukin-1beta , Mice , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Xanthones
Arq. bras. med. vet. zootec. (Online) ; 72(3): 647-654, May-June, 2020. ilus, tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1128504


The elastic cartilage is composed by chondroblasts and chondrocytes, extracellular matrix and surrounded by perichondrium. It has a low regeneration capacity and is a challenge in surgical repair. One of obstacles in engineering a structurally sound and long-lasting tissue is selecting the most appropriate scaffold material. One of the techniques for obtaining biomaterials from animal tissues is the decellularization that decreases antigenicity. In this work, alkaline solution was used in bovine ear elastic cartilages to evaluate the decellularization and the architecture of the extracellular matrix. The cartilages were treated in alkaline solution (pH13) for 72 hours and lyophilized to be compared with untreated cartilages by histological analysis (hematoxylin-eosin, Masson's trichrome and Verhoeff slides). Areas of interest for cell counting and elastic fiber quantification were delineated, and the distribution of collagen and elastic fibers and the presence of non-fibrous proteins were observed. The results demonstrated that the alkaline solution caused 90% decellularization in the middle and 13% in the peripheral region, and maintenance of the histological characteristics of the collagen and elastic fibers and non-fibrous protein removal. It was concluded that the alkaline solution was efficient in the decellularization and removal of non-fibrous proteins from the elastic cartilages of the bovine ear.(AU)

A cartilagem elástica é composta por condroblastos e condrócitos, matriz extracelular e envolta por pericôndrio. Possui uma baixa capacidade de regeneração e é um desafio em reparos cirúrgicos. Um dos obstáculos na engenharia de tecido estruturalmente sólido e de longa duração é a seleção do material de arcabouço mais adequado. Uma das técnicas para obtenção de biomateriais oriundos de tecidos animais é a descelularização, que diminui a antigenicidade. Neste trabalho, foi utilizada solução alcalina em cartilagem elástica auricular bovina para avaliar a descelularização e a arquitetura da matriz extracelular. As cartilagens foram tratadas em solução alcalina (pH13) durante 72 horas e liofilizadas, e comparadas com cartilagens não tratadas por análise histológica (hematoxilina-eosina, tricrômio de Masson e Verhoeff). Foram determinadas as áreas de interesse para contagem celular e quantificação de fibras elásticas, observada a distribuição de colágeno e fibras elásticas e a presença de proteínas não fibrosas. Os resultados demonstraram que a solução alcalina causou 90% de descelularização na região central e 13% na região periférica, manutenção das características histológicas do colágeno e fibras elásticas e remoção das proteínas não fibrosas. Concluiu-se que a solução alcalina foi eficiente na descelularização e retirada de proteínas não fibrosas de cartilagens elásticas da orelha de bovinos.(AU)

Biocompatible Materials , Chondrocytes , Tissue Engineering/veterinary , Elastic Cartilage , Extracellular Matrix , Cattle , Cartilage , Eosine Yellowish-(YS) , Alkalies
Article in Chinese | WPRIM | ID: wpr-879358


Osteoarthritis(OA) is a common clinical disease. The incidence of OA increases significantly with age, and the quality of life of patients is seriously affected. In the pathogenesis of OA, cartilage degeneration is the main cause. There are many long non-coding RNA (lncRNA) specifically expressed in osteoarthritis, which is closely related to the occurrence and development of osteoarthritis. Based on the latest research from 2014 to 2019, this paper summarizes the differential expression of lncRNA in osteoarthritis, the mechanism of lncRNA regulating chondrocyte function, and the mechanism of lncRNA regulating cartilage matrix metabolism. The fact that the expression of lncRNA is altered at different stages of OA development indicates that lncRNA can be developed forlife. The biomarkers and therapeutic targets can provide reference for the prevention, treatment and research of osteoarthritis.

Chondrocytes , Humans , Osteoarthritis/genetics , Quality of Life , RNA, Long Noncoding/genetics , Research
Article in Chinese | WPRIM | ID: wpr-828285


Knee osteoarthritis is the most common type of arthritis, which is manifested by the deformation and degeneration of articular cartilage and the discomfort of patients with joint pain, which affects the quality of life of patients and aggravates the medical burden of society. The pathogenesis of knee osteoarthritis is very complex. This paper reviews the inflammatory factors and signal pathways involved in knee osteoarthritis. It is found that most of the inflammatory factors involved are interleukin, such as IL-1 β, IL-6, IL-15, IL-17, IL-18, and tumor necrosis factors, such as TNF-α. These inflammatory factors aggravate knee osteoarthritisby activating corresponding pathways and promoting the release of inflammatory mediators. The inflammatory signaling pathways involved in knee osteoarthritis are complex. Notch pathway, Wnt pathway, SDF-1 / CXCR4 pathway, TLRs pathway, MAPKs pathway, hippo Yap pathway, OPG-RANK-RANKL pathway and TGF-β pathway are all involved in the regulation of knee osteoarthritis, and the pathways related to inflammatory mechanism are mainly MAPKs pathway and TLRs pathway. Different signaling pathways can cause the destruction of articular cartilage, promote the apoptosis of chondrocytes, and finally lead to the further imbalance of homeostasis in the knee joint. At the same time, the activation of signal pathway can promote the release of inflammatory factors, so under the cascade reaction of inflammatory factors and signal pathway, knee osteoarthritis is aggravating.

Cartilage, Articular , Chondrocytes , Humans , Interleukin-1beta , Osteoarthritis, Knee , Quality of Life , Signal Transduction
Article in Chinese | WPRIM | ID: wpr-827552


OBJECTIVE@#This study aimed to compare the cartilage regeneration of the stromal vascular fraction (SVF) cells and adipose-derived mesenchymal stem cells (ASCs) cocultured with chondrocytes seeded on the scaffolds.@*METHODS@#The cellular morphologies and proliferation capabilities on the scaffolds were evaluated. The scaffolds with the cocul-ture of ASCs/SVF and chondrocytes were implanted into the full thickness cartilage defective rabbit joints for 10 weeks.@*RESULTS@#The cells seeded into the scaffolds showed good adhesion and proliferation. Implantation with SVF and chondrocytes revealed desirable in vitro healing outcomes.@*CONCLUSIONS@#The SVF cells were better than ASCs in terms of the formation of cartilage matrix in a coimplantation model. Without in vitro expansion, the SVF cells are good cell sources for cartilage repair.

Adipose Tissue , Animals , Cartilage , Chondrocytes , Coculture Techniques , Rabbits , Regeneration
Journal of Biomedical Engineering ; (6): 1101-1108, 2020.
Article in Chinese | WPRIM | ID: wpr-879242


As a kind of mechanical effector cells, chondrocytes can produce a variety of physical and chemical signals under the stimulation of multiaxial load

Apoptosis , Cells, Cultured , Chondrocytes , Stress, Mechanical
Article in English | WPRIM | ID: wpr-881038


Due to the poor repair ability of cartilage tissue, regenerative medicine still faces great challenges in the repair of large articular cartilage defects. Quercetin is widely applied as a traditional Chinese medicine in tissue regeneration including liver, bone and skin tissues. However, the evidence for its effects and internal mechanisms for cartilage regeneration are limited. In the present study, the effects of quercetin on chondrocyte function were systematically evaluated by CCK8 assay, PCR assay, cartilaginous matrix staining assays, immunofluorescence assay, and western blotting. The results showed that quercetin significantly up-regulated the expression of chondrogenesis genes and stimulated the secretion of GAG (glycosaminoglycan) through activating the ERK, P38 and AKT signalling pathways in a dose-dependent manner. Furthermore, in vivo experiments revealed that quercetin-loaded silk protein scaffolds dramatically stimulated the formation of new cartilage-like tissue with higher histological scores in rat femoral cartilage defects. These data suggest that quercetin can effectively stimulate chondrogenesis in vitro and in vivo, demonstrating the potential application of quercetin in the regeneration of cartilage defects.

Animals , Cartilage/cytology , Chondrocytes/drug effects , Chondrogenesis/drug effects , Extracellular Matrix/metabolism , Quercetin/pharmacology , Rats , Signal Transduction/drug effects , Tissue Scaffolds
Rev. bras. ortop ; 54(6): 679-684, Nov.-Dec. 2019. tab, graf
Article in English | LILACS | ID: biblio-1057950


Abstract Objective To describe a postarthroscopic treatment classification system for acetabular chondral damage in the hip and to report the intraobserver and interobserver reliability of such classification. Methods This is a retrospective review of ninety-nine digital video recordings made during arthroscopic surgery. Patients who underwent arthroscopic treatment for femoroacetabular impingement and evaluated at the hip arthroscopy outpatient clinic between March 2015 and March 2016 were included in the study. Patients with a history of previous hip surgery, radiologic evidence of advanced osteoarthritis (Tönnis grade > 2), who underwent labral resection, or whose digital recordings were incomplete or of insufficient quality for adequate review were excluded. Two orthopedic surgeons, who did not participate in the surgery, independently reviewed the video recordings and classified the remaining acetabular cartilage using the post-treatment classification system. Intraobserver and interobserver analysis was then conducted using intraclass correlation coefficient (ICC). Results Excellent intraobserver reliability (ICC = 0.790; p < 0.001) and interobserver reliability (ICC = 0.882; p < 0.001) were observed. Both ICC values were statistically significant. Conclusion The posttreatment classification of the remaining acetabular cartilage has excellent intra and interobserver reliability.

Resumo Objetivo Descrever um sistema de classificação de tratamento pós-artroscópico para as lesões condrais acetabulares no quadril e relatar as confiabilidades intra e interobservador deste sistema. Métodos Esta é uma revisão retrospectiva de 99 gravações de vídeo digital realizadas durante artroscopia. Os pacientes submetidos a tratamento artroscópico para impacto femoroacetabular e avaliados no ambulatório de quadril entre março de 2015 e março de 2016 foram incluídos no estudo. Os pacientes com histórico de cirurgia anterior do quadril, evidência radiológica de osteoartrose avançada (Tönnis > 2), pacientes submetidos à ressecção labral ou cujas gravações digitais estavam incompletas ou de qualidade insuficiente para avaliação adequada foram excluídos. Dois ortopedistas, que não participaram da cirurgia, revisaram de forma independente as gravações de vídeo e classificaram a cartilagem acetabular remanescente usando o sistema de classificação pós-tratamento. A análise intra e interobservador foi então realizada utilizando o coeficiente de correlação intraclasse (CCI). Resultados Excelente confiabilidade intraobservador (CCI = 0,790; p < 0,001) e confiabilidade interobservador (CCI = 0,882; p < 0,001). Ambos os valores de CCI foram estatisticamente significativos. Conclusão a classificação pós-tratamento da cartilagem acetabular remanescente possui excelente confiabilidade intra e interobservador.

Humans , Osteoarthritis , Arthroscopy , Cartilage , Treatment Outcome , Chondrocytes/classification , Hip Injuries , Orthopedic Surgeons
Int. j. morphol ; 37(4): 1450-1455, Dec. 2019. tab, graf
Article in English | LILACS | ID: biblio-1040152


SUMMARY: The normal sequential development of the hip joint (HJ) was considered for the evaluation of the morphological and ultrastructural aspects of the joint cartilage of the proximal femoral head epiphysis in human fetuses between 16 to 31 weeks of intra uterine life (IUL). Twenty human fetuses were fixed in 10 % formalin solution. Fetuses were divided into 4 groups (n=5): Group 1 (G1): 16-19 weeks IUL; Group 2 (G2): 20-23 weeks IUL; Group 3 (G3): 24-27 weeks IUL and Group 4 (G4): 28-31 weeks IUL. The right moieties of the HJ were subjected to light microscopy to determine the chondrocyte area, volume, and density and the extracellular matrix (ECM) density. The collagen component in ECM was qualitatively evaluated using Safranin-O and picrosirius techniques under polarized light. The left portions were analyzed using scanning electron microscopy (SEM). The advance of age revealed a gradual increase in chondrocyte area and volume and in ECM density, and a decrease in chondrocyte density. The apparent prevalence of type II collagen fibers in G1 and type III collagen fibers in G4, as well as a balance between type I and III collagen fibers in G2 and G3 suggest a process of cartilaginous evolution and repair. The pantographic organization of the collagen fiber meshes from the depth to the cartilage surface of the femoral head suggests that the arcade collagen network architecture starts at the fetal stage, regardless of the compressive forces applied. The morphological data may contribute not only to a better understanding of the maturation and cartilage organization in this area but also to serve as a theoretical basis for aspects related to diseases and joint malformations.

RESUMEN: El desarrollo secuencial normal de la articulación de la cadera (AC) se consideró para la evaluación de los aspectos morfológicos y ultraestructurales del cartílago articular de la epífisis proximal y de la cabeza femoral en fetos humanos entre 16 y 31 semanas de vida intrauterina (SVIU). Veinte fetos humanos fueron fijados en solución de formalina al 10 %. Los fetos se dividieron en 4 grupos (n = 5): Grupo 1 (G1), 1619 semanas de IUL; Grupo 2 (G2), 20-23 semanas SVIU; Grupo 3 (G3), 24-27 semanas SVIU y Grupo 4 (G4), 28-31 semanas SVIU. Las muestras derechas de la AC se sometieron a microscopía óptica para determinar el área, el volumen y la densidad de los condrocitos y la densidad de la matriz extracelular (MEC). El componente de colágeno en la MEC se evaluó cualitativamente utilizando técnicas de safranina-O y picrosirius bajo luz polarizada. Las muestras de la AC izquierda se analizaron utilizando microscopía electrónica de barrido (MEB). El avance de la edad reveló un aumento gradual en el área y el volumen de los condrocitos y en la densidad de la MEB, y una disminución en la densidad de los condrocitos. La aparente prevalencia de las fibras de colágeno tipo II en G1 y tipo III en G4, así como el equilibrio entre las fibras de colágeno tipo I y III en G2 y G3 sugieren un proceso de evolución y reparación cartilaginosa. La organización pantográfica de las mallas de fibra de colágeno desde la profundidad a la superficie del cartílago de la cabeza femoral sugiere que la arquitectura de la red de colágeno comienza en la etapa fetal, independientemente de las fuerzas compresivas aplicadas. Los datos morfológicos pueden contribuir no solo a una mejor comprensión de la organización de la maduración y el cartílago en esta área, sino también servir de base teórica para los aspectos relacionados con enfermedades y malformaciones articulares.

Humans , Fetus , Hip Joint/ultrastructure , Microscopy, Electron, Scanning , Collagen/ultrastructure , Chondrocytes/ultrastructure , Extracellular Matrix , Hip Joint/embryology
Rev. bras. ortop ; 54(5): 549-555, Sept.-Oct. 2019. tab, graf
Article in English | LILACS | ID: biblio-1057930


Abstract Objective To evaluate the clinical and radiological benefits of intra-articular exogenous hyaluronic acid for the treatment of chondral patellar injury. Method Randomized clinical trial with 70 patients divided into 2 groups: those submitted to physical therapy for 3 months, and those submitted to physical therapy associated with the intra-articular administration of 2 mL of hyaluronic acid for the same period, who had anterior knee pain and patellar cartilage injury of grades II or III with no significant bone abnormalities. The functional scores and the characteristics of the physical and imaging exams were evaluated before and 3 and 6 months after the treatment. Result The average age of the patients was 32 ± 7.6 years. Patients from the hyaluronic acid group had better Kujala et al and Lysholm scores, and lower pain scores after 3 and 6 months of treatment when compared to the control group. The incidence of positive Clarke maneuver was lower in the treated group, but there was no difference in the magnetic resonance imaging classification. Conclusion Patients with patellar chondropathy of grades II or III treated with hyaluronic acid and physical therapy had less pain (visual analogue scale, VAS), and better functional results in the Lysholm and Kujala et al questionnaires after 3 and 6 months of treatment compared to patients undergoing physical therapy alone. In addition, the number of cases with a negative Clarke maneuver was larger in the treated group after 6 months of treatment.

Resumo Objetivo Avaliar os benefícios clínicos e radiológicos do uso do ácido hialurônico exógeno intra-articular para o tratamento da lesão condral da patela. Método Ensaio clínico randomizado com 70 pacientes divididos em dois grupos: o de tratamento fisioterápico por 3 meses, e o de tratamento fisioterápico associado à aplicação de 2 ml de ácido hialurônico intra-articular pelo mesmo período, composto por pacientes com dor na região anterior do joelho e lesão de graus II ou III da cartilagem da patela, sem anormalidades ósseas significativas. Foram avaliados os escores funcionais e as características do exame físico e de imagem antes, e após 3 e 6 meses de tratamento. Resultado A idade média dos pacientes foi de 32 ± 7,6 anos. Os pacientes do grupo submetido à aplicação de acido hialurônico apresentaram melhores escores de Kujala et al e de Lysholm, e menor pontuação de dor após 3 e 6 meses de tratamento quando comparados ao grupo controle. A manobra de Clarke positiva foi menor no grupo em que foi feita a aplicação do ácido, mas não houve diferença na classificação da imagem obtida pela ressonância magnética. Conclusão Pacientes com condropatia patelar de graus II ou III do joelho tratados com ácido hialurônico e fisioterapia apresentaram menos dor (escala visual analógica, EVA), e melhores resultados funcionais nos questionários de Lysholm e de Kujala et al após 3 e 6 meses de tratamento quando comparados com os pacientes submetidos apenas à fisioterapia. Além disso, estes pacientes apresentaram manobra de Clarke negativa em maior número após 6 meses de tratamento.

Humans , Male , Female , Adolescent , Adult , Middle Aged , Pain , Patella , Cartilage Diseases , Extravehicular Activity , Chondrocytes , Hyaluronic Acid
Arq. bras. med. vet. zootec. (Online) ; 71(2): 509-520, mar.-abr. 2019. graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1011276


The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Animals , Female , Rats , Caffeine , Cartilage, Articular/drug effects , Chondrocytes/drug effects , Chondrogenesis/drug effects
Artrosc. (B. Aires) ; 26(3): 83-87, 2019.
Article in Spanish | LILACS, BINACIS | ID: biblio-1048246


Introducción: En la actualidad existen diferentes métodos y técnicas de preservación articular. La utilización de una matriz de atelocolágeno combinada con microperforaciones otorga un soporte adecuado para la inducción de la condrogénesis a partir de las células mesenquimales provenientes de la médula ósea. El objetivo de nuestro trabajo es describir la técnica quirúrgica y presentar los resultados de una serie de pacientes con lesiones condrales severas, tratados con microperforaciones asociado a una matriz de atelocolágeno. Material y Método: Se evaluaron los pacientes intervenidos quirúrgicamente por lesión de cartílago grado IV de más de 3 cm2 a los que se le aplicó matriz de atelocolágeno combinado con microperforaciones. El mínimo seguimiento fue de 24 meses. En pacientes con deseje o inestabilidad asociada se realizaron procedimientos combinados en el mismo acto quirúrgico. Describimos la técnica quirúrgica, resultados funcionales pre y postoperatorios con las escalas de Lysholm, IKDC y Escala Visual Análoga (EVA) del dolor fueron. Se realizó una evaluación radiográfica. Analizamos las complicaciones del procedimiento. Resultado: Fueron operados 12 pacientes. A uno se le realizó un reemplazo articular de su rodilla a los 10 meses de la cirugía y fue considerado falla con finalización del seguimiento. Once fueron evaluados clínicamente, nueve hombres y dos mujeres, con una edad promedio de 48 años y seguimiento promedio de 34 meses. Ocho procedimientos en cóndilo interno, 2 en cóndilo externo y 4 en tróclea. La mediana de la escala de IKDC pre/post operatorio fue 41/55 (p 0.016), Lysholm 35/82 (p 0.004) y EVA 9/3 (p 0.002). La evaluación radiológica no evidenció cambios degenerativos. Se registró 1 artrofibrosis post operatoria. Conclusión: En nuestra serie, el tratamiento con atelocolágeno combinado con microperforaciones mejoró la clínica de los pacientes con lesión severa del cartílago articular de rodilla. Tipo de trabajo: Serie de casos Nivel de Evidencia: IV

ntroduction: Different surgical approaches are currently available to treat knee chondral defects. The technique used in this article combines microfractures with the use of an injectable atelocollagen matrix (Cartifill). The matrix covers the defect and improves the mechanical stability of the blood clot and maintains the chondrogenic progenitor cells and growth factors in the defective area. The aim of our study is to evaluate and describe the results in a series of patients treated with atelocollagen matrix and microfractures. Material and Methods: All patients treated with atelocollagen matrix due to a cartilage lesion with a minimum follow-up of 24 months were evaluated. Patients undergoing associated surgeries (osteotomies, meniscectomies, mosaicplasty, ligament reconstruction) in the same surgical procedure were included in the study. Clinical function was assessed before and after surgery with the International Knee Documentation Committee (IKDC), the Lysholm score and the Visual Analogue Scale (VAS). Radiographic control was requested according to availability. Results: Twelve patients met the inclusion criteria. Three women. Average age of 50 years. Eight applications in medial condyle, 2 in lateral condyle and 4 in trochlea. One post-operative arthrofibrosis was recorded. One of the patients underwent an articular replacement of his knee 10 months after the surgery with finalization of follow-up. The pre / post-operative average was 39/52 (IKDC), 37/76 (Lysholm) and 8.5 / 3.5 (VAS). Conclusion: In our series, atelocollagen matrix combined with microfractures improved the clinical symptoms of patients with severe knee articular cartilage injury. However, a better selection of patients who require this procedure should be applied in future interventions. Type of Study: Case Series. Level of Evidence: IV

Adult , Middle Aged , Arthroscopy/methods , Cartilage, Articular/surgery , Cartilage, Articular/injuries , Collagen/therapeutic use , Chondrocytes/transplantation , Tissue Engineering/methods , Knee Injuries/surgery
Article in English | WPRIM | ID: wpr-763051


Osteoarthritis is a chronic degenerative articular disorder. Formation of bone spurs, synovial inflammation, loss of cartilage, and underlying bone restructuring have been reported to be the main pathologic characteristics of osteoarthritis symptoms. The onset and progression of osteoarthritis are attributed to various inflammatory cytokines in joint tissues and fluids that are produced by chondrocytes and/or interact with chondrocytes, as well as to low-grade inflammation in intra-articular tissues. Disruption of the equilibrium between the synthesis and degradation of the cartilage of the joint is the major cause of osteoarthritis. Hence, developing a promising pharmacological tool to restore the equilibrium between the synthesis and degradation of osteoarthritic joint cartilage can be a useful strategy for effectively managing osteoarthritis. In this review, we provide an overview of the research results pertaining to the search for a novel candidate agent for osteoarthritis management via restoration of the equilibrium between cartilage synthesis and degradation. We especially focused on investigations of medicinal plants and natural products derived from them to shed light on the potential pharmacotherapy of osteoarthritis.

Biological Products , Cartilage , Chondrocytes , Cytokines , Drug Therapy , Inflammation , Joints , Osteoarthritis , Osteophyte , Plants, Medicinal
Asian Spine Journal ; : 361-367, 2019.
Article in English | WPRIM | ID: wpr-762957


STUDY DESIGN: Experimental study. PURPOSE: To determine whether epidural fat (EF) tissue contains mesenchymal stem cells (MSC). OVERVIEW OF LITERATURE: Spine surgeons are unaware of the contents of EF tissue and the reason for its presence between the ligamentum flavum and the dura mater; therefore, EF tissues are routinely eliminated during surgical procedures. However, EF removal causes certain postoperative problems, such as post-laminectomy syndrome. We hypothesized that the EF tissue may play a significant supportive role for the neural structures and other nearby conditions. METHODS: EF tissues were obtained from consenting patients (n=3) during posterior decompression surgery of the lumbar spine. The primary cells were isolated and cultured as per previously described methods with some modifications, and the cell morphology and cumulation were examined. Thereafter, reverse transcription–polymerase chain reaction (RT-PCR), a fluorescence-activated cell sorting (FACS) analysis, and differentiation potency for differentiation into osteoblasts, chondroblasts, and adipocytes were investigated to identify whether the cells derived from EF are MSC. RESULTS: The cells from the EF tissue had a fibroblast or neuron-like morphology that persisted until the senescence at p18. MSC-specific genes, such as OCT4, SOX2, KLF4, MYC, and GAPDH were expressed in the RT-PCR study, while MSC-specific surface markers such as CD105, CD90, and CD73 were exhibited in the FACS analysis. The differentiation properties of EF-MSC for differentiation into the three types of cells (osteoblast, chondroblast, and adipocyte) were also confirmed. CONCLUSIONS: Based on the cell culture, FACS analysis, RT-PCR analysis, and differentiation potent outcomes, all the features of the cells corresponded to MSC. This is the first study to identify EF-MSC derived from the EF tissue.

Adipocytes , Aging , Cell Culture Techniques , Chondrocytes , Decompression , Dura Mater , Fibroblasts , Flow Cytometry , Humans , Ligamentum Flavum , Mesenchymal Stem Cells , Osteoblasts , Spine , Surgeons