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ABSTRACT Objective: To test the hypothesis that undenatured type II collagen (UC-II) relieves pain, quality of life, and joint function in women aged from 60 to 80 years with knee osteoarthritis. Methods: 53 patients in the UC-II treatment group (for 90 days) and 52 in the control group (without UC-II) were evaluated at 1, 30, and 90 days regarding health-related quality of life, pain, and function with questionnaires, anthropometric data, alignment, range of motion, and radiographic analysis. Results: Quality of life increased significantly in the Physical domain in the treatment vs control group. Also, there was a difference between the first and the last evaluation on the pain visual analog scale (−3.8 ± 1.8 versus −1.3 ± 2.0) and on the WOMAC score (−9.5 ± 11.9 versus −1.3 ± 11.1). No variation in the temporal evolution of the Mental domain was found. Conclusion: Pain, joint stiffness, and quality of life (Physical domain) improved with the inclusion of UC-II for 90 days to the therapeutic toolbox for knee osteoarthritis in individuals aged 60 to 80 years. Level of evidence II, Comparative Prospective Study.
RESUMO Objetivo: Testar a hipótese de que o colágeno não hidrolisado tipo II (UC-II) melhora a dor, qualidade de vida e função articular de indivíduos entre 60 e 80 anos com osteoartrite (OA) de joelho. Métodos: Cinquenta e três pacientes do grupo tratamento com UC-II (por 90 dias) e 52 do grupo controle (GC - sem UC-II) foram avaliados no tempo 0, 30 e 90 dias quanto à qualidade de vida em saúde, dor e função com os questionários, além de dados antropométricos, alinhamento, amplitude de movimento e análise radiográfica. Resultados: A qualidade de vida aumentou significantemente no domínio PCS no grupo tratamento versus controle. Houve ainda diferença entre a primeira e última avaliação na dor pela escala visual analógica (−3.8 ± 1.8 versus −1.3 ± 2.0) e no escore WOMAC (−9.5 ± 11.9 versus −1.3 ± 11.1). Não houve variação na evolução temporal do domínio MCS. Conclusão: Dor, rigidez articular e qualidade de vida (domínio físico) melhoram com a inclusão do UC-II por 90 dias ao arsenal terapêutico na OA do joelho em indivíduos de 60 a 80 anos. Nível de Evidência II, Estudo Prospectivo Comparativo.
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BACKGROUND: Preliminary studies have shown that numerous antioxidants exhibit antiarthritic effects due to their inhibition on inflammatory factors. With the antioxidant and anti-inflammatory abilities whether protocatechuic acid is effective in the treatment of osteoarthritis has never been reported. OBJECTIVE: To investigate the biological effect of protocatechuic acid on interleukin-1β induced chondrocyte including phenotype and cellular metabolism in vitro, thereby providing a potential agent in osteoarthritis treatment, METHODS: The chondrocytes of neonatal rat femur were collected, intervened by 10 mg/L interleukin-1β to establish the degenerative model, and treated by 10, 30 and 50 mg/L protocatechuic acid. The study was approved by the Laboratory Animal Ethical Committee of Guangxi Medical University in October 2017, with the approval No. 201710008. RESULTS AND CONCLUSION: Protocatechuic acid effectively promoted chondrocyte growth in the range of 10-50 mg/L, while the dose of 30 mg/L was the strongest. Protocatechuic acid also enhanced the synthesis of the extracellular matrix and the mRNA expression of aggrecan, collagen II and Sox9, and downregulating the expression of the matrix metalloproteinase 13 (a marker of inflammatory factor). To conclude, protocatechuic acid exerts a positive effect on the proliferation and phenotypic maintenance of articular chondrocytes, providing reference for its use in the treatment of osteoarthritis and repair of degenerative articular cartilage in vivo.
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BACKGROUND: Osteoporotic fracture combined with type 2 diabetic mellitus in female patients is often accompanied by dyslipidemia and hyperglycemia. In addition to insulin treatment, statins are often prescribed for combination therapy, but the combined effect of these two drugs on fracture healing has not been reported in such patients. OBJECTIVE; To investigate the effect of lovastatin combined with insulin on the fracture healing of bilateral ovariectomized rats suffering from type 2 diabetic mellitus. METHODS: The study was approved by the Ethical Committee of North China University of Science and Technology. Thirty-two female Sprague-Dawley rats were divided into control, diabetic ovariectomized, insulin and combined groups. A model of type 2 diabetes and osteoporosis fracture was established in all rats except for the control group. At 7 days after the type 2 diabetic model was successfully established by injection of streptozotocin, the rats in the insulin and combined groups received the subcutaneous injection of insulin (2-4 U in the morning, and 4-6 U in the evening) until the end of the experiment. The rats in the combined group were given 20 mg/kg lovastatin via gavage daily, and those in the other two groups were not treated. All rats were sacrificed at 3 weeks after fracture. Radiographic, clinical and histomorphometric detections of the callus were performed. The expression levels of bone morphogenetic protein 2, vascular endothelial growth factor and collagen type II were detected. All above results were used to analyze the fracture healing in each group. RESULTS AND CONCLUSION: (1)The radiographic score, micro-CT index, histologic score and the expression levels of vascular endothelial growth factor and collagen type II in the diabetic ovariectomized group were significantly poorer than those in the control group (P < 0.05). (2) All above indexes in the insulin group were significantly improved compared with the diabetic ovariectomized group (P < 0.05), which promoted the fracture healing of model rats. (3) After combined with lovastatin, although the expression levels of vascular endothelial growth factor and bone morphogenetic protein 2 in the callus were significantly increased (P < 0.05), there was no significant improvement in the radiographic appearance and microstructure of the callus.
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BACKGROUND: The establishment of coculture system combined with physical factors and scaffold materials and the Induction of cytokines have become the focus of chondrogenlc differentiation of bone marrow mesenchymal stem cells. OBJECTIVE: To observe the effect of bone morphogenetic protein 7 combined with porous tantalum on chondrogenlc differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats (provided by Beijing Huafukang Biology) were Isolated and cultured. Group Intervention: (1) in the experimental group, porous tantalum tablet was added, while In the control group, porous tantalum tablet was not added. At 5 days after culture, cell growth on the surface of porous tantalum tablet was observed by phalloidin staining. At 1, 3,5, and 7 days after culture, CCK-8 method was used to detect cell proliferation. (2) Group A was added with chondrocyte Inducer; group B with chondrocyte Inducer and bone morphogenetic protein 7; group C with domestic porous tantalum material and chondrocyte Inducer; group D with domestic porous tantalum material and chondrocyte Inducer and bone morphogenetic protein 7. At 7,14 and 21 days after culture, the levels of type II collagen, SRY type high mobility group protein and matrix metalloproteinase-13 secreted by cells In each group was detected by ELISA. Western blot assay was used to detect the expression of type II collagen, SRY type high mobility group protein and matrix metalloproteinase-13. This study was approved by the Animal Experimental Ethics Committee of North China University of Science and Technology. RESULTS AND CONCLUSION: (1) The phalloidin staining results showed that bone marrow mesenchymal stem cells grew well on and around the porous tantalum surface. (2) At 3 and 5 days after culture, the proliferation of bone marrow mesenchymal stem cells was slower In the experimental group than in the control group (P 0.05). (3) At 7,14 and 21 days, the expression of type II collagen and SRY high mobility group protein Increased gradually among groups A, B, C and D (P 0.05). At 21 days, there was no significant difference among groups A, B, C and D (P > 0.05). (4) Western blot assay showed that at 7,14 and 21 days after culture, the expression level of type II collagen and SRY high mobility group protein Increased gradually In groups A, B, C and D (P 0.05). (5) The results showed that bone morphogenetic proteln-7 combined with domestic porous tantalum could Induce cartilage differentiation of bone marrow mesenchymal stem cells, facilitate the expression of type II collagen and SRY high mobility group protein, and Inhibit the expression of matrix metalloproteinase-13.
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BACKGROUND/OBJECTIVES: We investigated the anti-osteoarthritic effects of deer bone extract on the gene expressions of matrix metalloproteinases (MMPs) and collagen type II (COL2) in interleukin-1β-induced osteoarthritis (OA) chondrocytes. MATERIALS/METHODS: Primary rabbit chondrocytes were treated as follows: CON (PBS treatment), NC (IL-1β treatment), PC (IL-1β + 100 µg/mL glucosamine sulphate/chondroitin sulphate mixture), and DB (IL-1β + 100 µg/mL deer bone extract). RESULTS: The results of the cell viability assay indicated that deer bone extract at doses ranging from 100 to 500 µg/mL inhibits cell death in chondrocytes induced by IL-1β. Deer bone extract was able to significantly recover the mRNA expression of COL2 that was down-regulated by IL-1β (NC: 0.79 vs. DB: 0.87, P < 0.05) and significantly decrease the mRNA expression of MMP-3 (NC: 2.24 vs. DB: 1.75) and -13 (NC: 1.28 vs. DB: 0.89) in OA chondrocytes (P < 0.05). CONCLUSIONS: We concluded that deer bone extract induces accumulation of COL2 through the down-regulation of MMPs in IL-1β-induced OA chondrocytes. Our results suggest that deer bone extract, which contains various components related to OA, including chondroitin sulphate, may possess anti-osteoarthritic properties and be of value in inhibiting the pathogenesis of OA.
Subject(s)
Cell Death , Cell Survival , Chondrocytes , Chondroitin , Collagen Type II , Collagen , Deer , Down-Regulation , Gangliosides , Gene Expression , Glucosamine , Matrix Metalloproteinases , Osteoarthritis , RNA, MessengerABSTRACT
Objective To investigate the effects of compound cornu cervi degelatinatum on the expression of related inflammatory cytokines and pathological changes of the synovium in mice with collagen-induced arthritis (CIA). MethodsThe CIA model of Balb/c mice was established. The mice were randomly divided into 5 groups: blank control group, model control group, low-dose group ( C1 ) , high-dose group ( C2 ) and glucosidorum tripterygll totorum group D. After the first immunization in mice, we observed the general condition and lesions of the fore and hind jaws. After the second immunization, the mice in groups C1 and C2 were orally administered with compound cornu cervi degelatinatum at the doses of 2. 5 and 5 g·kg-1 body weight per day for 4 weeks, and meanwhile those in group D were administrated with glucosidorum tripterygll totorum at the dose of 13. 6 mg·kg-1 body weight per day. The serum of the mice was collected to detect the levels of TNF-αand IL-4 by ELISA. Ankle joints were harvested, and the pathological changes of synovial tissues were observed under light microscope. Results As compared with the model control group, the level of TNF-αin the treatment groups was significantly decreased, while the level of IL-4 was elevated in group C1. Histological pathology of ankle joints demonstrated that the synovium of the CIA mice were hyperplastic and the synovial tissues were markedly ameliorated in treatment groups. Conclusion The compound cornu cervi degelatinatum can relieve redness and swelling in mice with CIA. The mechanism may be related to the inhibition of pro-inflammatory cytokine TNF-α and regulation of anti-inflammatory cytokine IL-4.
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OBJECTIVE: To investigate the feasibility of cultivated and wild total secoiridoid glucoside in treating osteoarthritis (OA) by observing the effects of total secoiridoid glucoside on interleukin 1β(IL-1β) induced chondrocytes of rat in vitro.
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BACKGROUND:Compared with other sources of mesenchymal stem cells, synovial-derived mesenchymal stem cells have significant characteristics of chondrogenesis and cloning. Therefore, synovial-derived mesenchymal stem cells are one of the most promising seed cells in cartilage tissue engineering. OBJECTIVE:To isolate and culture synovial-derived mesenchymal stem cells of Sprague-Dawley rats, identify the multipotential differentiation and the potential ability of chondrogenic differentiation in three-dimensional culture condition. METHODS:The synovium tissue was harvested from Sprague-Dawley rats. The synovial-derived mesenchymal stem cells were isolated with typeⅠcol agen enzyme digestion method and cultured in vitro. The passage 3 cells were detected with giemsa staining, the cellcycle, adipogenic and osteogenic differentiation were determined. The passage 3 cells were centrifuged as pel ets and cultured in the chondriogenic medium for 21 days. And the pel ets were examined by toluidine blue staining, typeⅡcol agen immunohistochemical staining and RT-PCR. RESULTS AND CONCLUSION:The mesenchymal stem cells isolated from the synovium tissue of rats have the characteristics of mesenchymal stem cells, and exhibit fibroblast-like morphology after cultured in vitro. The multilineage differentiation potentials were also revealed. After the cellwere cultured in chondrogenic medium for 21 days, chondroid tissue was found, type II col agen and aggrecan could be detected positively by toluidine blue staining, typeⅡcol agen immunohistochemical staining, and expressed by RT-PCR examination. Therefore, synovial mesenchymal stem cells have a chondrogenic differentiation potential.
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BACKGROUND:Nowadays, growth factors are commonly used to induce bone marrow mesenchymal stem cells. However, this is a high-cost method with a great amount of growth factors. In addition, the chondrogenic potential of bone marrow mesenchymal stem cells wil decrease significantly with increasing times of culture. OBJECTIVE:To observe the directed differentiation of bone marrow mesenchymal stem cells co-cultured with human synovial fluid. METHODS:Human bone marrow mesenchymal stem cells were isolated and cultured by adherence screening method. The synovial fluid of the knee was aspirated from healthy volunteers by aseptic operation. Passage 3 human bone marrow mesenchymal stem cells were co-cultured with the fol owing media:synovial fluid+complete medium;synovial fluid+bone marrow mesenchymal stem cells+complete medium;bone marrow mesenchymal stem cells+complete medium. The morphology and growth of the cells were observed under an inverted microscope every day. At days 7, 14 and 21 of induction, toluidine blue staining and immunocytochemical staining were performed. RESULTS AND CONCLUSION:After co-culture with human synovial fluid, human bone marrow mesenchymal stem cells proliferated slowly, and varied from fusiform to oval or polygonal;toluidine blue and col agen II staining were positive. These findings indicate that the synovial fluid has a positive role in the chondrogenic differentiation of bone marrow mesenchymal stem cells. The synovial fluid may contain substances that promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.
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Dentro de las enfermedades autoinmunes, la Artritis Reumatoide es una de las más estudiadas, dada su alta prevalencia. Numerosos estudios han reportado como posible mecanismo gatillante la existencia de una respuesta inmune contra el Colágeno tipo II. El presente trabajo busca contribuir al conocimiento y significación de este proceso, a través de la determinación de diferentes auto-anticuerpos en sujetos con Artritis Reumatoide (diagnosticados bajo criterios del American College of Rheumatology) y en sujetos con artralgias inespecíficas en comparación con sujetos normales. Para esto, se realizó un estudio descriptivo, de corte transversal, en el que se determinó la presencia, en sueros, de anticuerpos anti-colágeno tipo II y de anticuerpos anti-Péptidos Cíclicos Citrulinizados (CCP) por ELISA y de Anticuerpos anti-sinovial por Inmunofluorescencia Indirecta (IFI) con métodos previamente estandarizados de acuerdo a procedimientos convencionales. No se encontró diferencia significativa entre los promedios de los niveles de los anticuerpos anti-colágeno tipo II en los 3 grupos; sin embargo, la distribución de los títulos de anticuerpos presenta, en los tres grupos, curvas de frecuencia de tipo bi-modal cuyos perfiles son totalmente diferentes entre ellos. En este orden, se identificó un subgrupo conformado por el 20 por ciento del grupo con Artritis Reumatoide quienes presentaban anticuerpos anti-colágeno tipo II con niveles muy altos, por encima de las dos desviaciones estándar; dichos sujetos correspondían a adultos jóvenes (edad promedio 41 años) que no recibieron tratamiento inmunosupresor. No se encontró ninguna correlación entre los niveles de anticuerpos anti-colágeno tipo II, los anticuerpos anti-CCP, y los de anticuerpos anti-sinovial. Se discute la significación de estos hallazgos.
Between the Autoimmune Diseases, Rheumatoid Arthritis (RA) is one of the most studied, due to its high prevalence. Several studies have reported as possible shooting mechanism the existence of an immune response against Collagen type II. The present work look for contributing to the knowledge and meaning of the autoimmune process through determinations of different serum antibodies in subjects with Rheumatoid Arthritis (diagnosed under the American College of Rheumatology criteria) and in subjects with unspecific arthralgia in comparison with normal subjects. For this purpose, a transversal descriptive study was carried out, determining the presence of anti-collagen type II antibodies, anti-CCP (Citrulinated Cyclic Peptides) antibodies and anti-synovial antibodies by indirect immuno fluorescence (IFI) in sera, using previously standardized methods according to conventional procedures. No significant differences was found between the averages of the levels of anti-collagen type II in the 3 groups; however, the distribution of the antibody titles shows, in the three groups, curves of frequency of a bi-modal type, with profiles totally different between them. In this order, inside the group with Rheumatoid Arthritis it was identified a subgroup formed by the 20 percent of the subjects that presents very high levels of anti-collagen type II Antibodies (over the two standard deviations), made by young adults (average age 41 years) that did not receive inmunosuppresive treatment. It was not found relationship between the levels of anti-collagen type II antibodies, anti-CCP antibodies and anti-synovial antibodies. The significance of these findings is discussed.
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Antibodies/analysis , Arthritis, RheumatoidABSTRACT
2-deoxy-D-glucose (2DG) is known as a synthetic inhibitor of glucose. 2DG regulates various cellular responses including proliferation, apoptosis and differentiation by regulation of glucose metabolism in cancer cells. However, the effects of 2DG in normal cells, including chondrocytes, are not clear yet. We examined the effects of 2DG on dedifferentiation with a focus on the beta-catenin pathway in rabbit articular chondrocytes. The rabbit articular chondrocytes were treated with 5 mM 2DG for the indicated time periods or with various concentrations of 2DG for 24 h, and the expression of type II collagen, c-jun and beta-catenin was determined by Western blot, RT-PCR, immunofluorescence staining and immunohistochemical staining and reduction of sulfated proteoglycan synthesis detected by Alcain blue staining. Luciferase assay using a TCF (T cell factor)/LEF (lymphoid enhancer factor) reporter construct was used to demonstrate the transcriptional activity of beta-catenin. We found that 2DG treatment caused a decrease of type II collagen expression. 2DG induced dedifferentiation was dependent on activation of beta-catenin, as the 2DG stimulated accumulation of beta-catenin, which is characterized by translocation of beta-catenin into the nucleus determined by immunofluorescence staining and luciferase assay. Inhibition of beta-catenin degradation by inhibition of glycogen synthase kinase 3-beta with lithium chloride (LiCl) or inhibition of proteasome with z-Leu-Leu-Leu-CHO (MG132) accelerated the decrease of type II collagen expression in the chondrocytes. 2DG regulated the post-translational level of beta-catenin whereas the transcriptional level of beta-catenin was not altered. These results collectively showed that 2DG regulates dedifferentiation via beta-catenin pathway in rabbit articular chondrocytes.
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Animals , Rabbits , Cartilage, Articular/cytology , Cell Dedifferentiation/drug effects , Cell Nucleus/drug effects , Chondrocytes/cytology , Deoxyglucose/pharmacology , Endoplasmic Reticulum/drug effects , Glycogen Synthase Kinase 3/metabolism , Mutant Proteins/metabolism , Protein Transport/drug effects , Proteoglycans/metabolism , Signal Transduction/drug effects , beta Catenin/metabolismABSTRACT
Cyclooxygenase-2 (COX-2) is known to modulate bone metabolism, including bone formation and resorption. Because cartilage serves as a template for endochondral bone formation and because cartilage development is initiated by the differentiation of mesenchymal cells into chondrocytes (Ahrens et al., 1977; Sandell and Adler, 1999; Solursh, 1989), it is of interest to know whether COX-2 expression affect chondrocyte differentiation. Therefore, we investigated the effects of COX-2 protein on differentiation in rabbit articular chondrocyte and chick limb bud mesenchymal cells. Overexpression of COX-2 protein was induced by the COX-2 cDNA transfection. Ectopic expression of COX-2 was sufficient to causes dedifferentiation in articular chondrocytes as determined by the expression of type II collagen via Alcian blue staining and Western blot. Also, COX-2 overexpression caused suppression of SOX-9 expression, a major transcription factor that regulates type II collagen expression, as indicated by the Western blot and RT-PCR. We further examined ectopic expression of COX-2 in chondrifying mesenchymal cells. As expected, COX-2 cDNA transfection blocked cartilage nodule formation as determined by Alcian blue staining. Our results collectively suggest that COX-2 overexpression causes dedifferentiation in articular chondrocytes and inhibits chondrogenic differentiation of mesenchymal cells.
Subject(s)
Animals , Chick Embryo , Rabbits , Cartilage, Articular/cytology , Cell Differentiation , Cells, Cultured , Chondrocytes/cytology , Chondrogenesis , Collagen Type II/metabolism , Cyclooxygenase 2/biosynthesis , Interleukin-1beta/pharmacology , Mesenchymal Stem Cells/cytology , SOX9 Transcription Factor/metabolismABSTRACT
[Objective] To examine the specific gene and protein expression of primary and passaged chondrocytes in normal oxygen tension and hypoxia.[Methods]Articular chondrocytes were isolated from limb joint cartilage of C57BL/6 mice(3~5 days).Primary chondrocytes(P0)and passaled chondrocytes(P1,P2)were cultured in normal oxygen tension and hypoxia respectively for two days.The mRNA levels of collagen II,aggrecan,sox9,Indian hedgehog(ihh),parathyroid hormone-related protein(PTHrP),bone morphogenetic protein(BMP)-4 and wnt5a were examined with RT-PCR,and protein levels of collagen II,aggrecan were examined with immunohistochemistry and toluidine blue staining.[Results]During the culture of primary chondrocyte,the expression of collagen II and aggrecan increased under hypoxia,mRNA levels of collagen II,wnt5a increased and ihh decreased at the same time.The mRNA levels of special genes and differentiation related genes of passaged chondrocytes were not altered under hypoxia.[Conclusion]Protein and mRNA level of Collagen II of primary chondrocyte under hypoxia increased.It may be regulated by chondrocyte differentiation gene ihh,and wnt5a.The chondrocyte phenotype could not be resumed under hypoxia through short-term monolayer culture in vitro.
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BACKGROUND AND OBJECTIVES:Altered immunity against one's own tissue, especially inner ear, is in part responsible for the development of Meniere's disease (MD). Since immunologic screening test results are usually nonspecific, even if tested for those patients suspicious of autoimmune inner ear disease, they do not seem to provide valuable information. The aim of this study is to evaluate the validity of immunologic screening laboratory tests for MD patients with type II collagen (CII) autoimmunity and to discuss the diagnostic role in this localized immunologic disease. MATERIALS AND METHOD:Thirty-six patients of MD in which immunologic screening laboratory test result was available were included in this study and their clinical features were described according to the criteria of AAO-HNS (1995). They were divided into two groups, CII(+) (N=8) or CII(-) (N=28) according to the presence of anti-CII antibody determined by ELISA method as described earlier. Rheumatoid factor (RF), Fluorescent antinuclear antibody (FANA), immunoglobulin G, M and A, and complement 3 and 4 were included for immunologic screening test. Individual test and clinical features were compared between groups. RESULTS:Any single test did not show significant correlation between groups. But RF, total IgG and the proportion of patients more than at least one marker are higher in CII(+) group with borderline significance. CONCLUSION:Higher positive rate of immunologic screening test may support the immunologic involvement in CII(+) group. However the role of this screening test seems to be limited in a localized disease like MD compared systemic immunologic disorder, such as rheumatoid arthritis.
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Humans , Antibodies, Antinuclear , Arthritis, Rheumatoid , Autoimmunity , Collagen Type II , Complement C3 , Ear, Inner , Enzyme-Linked Immunosorbent Assay , Immune System Diseases , Immunoglobulin G , Labyrinth Diseases , Mass Screening , Meniere Disease , Rheumatoid FactorABSTRACT
The purpose of these studies was an establishment of human auricular chondrocyte cell line using retrovirus mediated v-myc transfer, characterizing the human auricular chondrocyte cell line by type II collagen mRNA expression and transplantation of human auricular cell line into immunological incompetent nude mice to establish neocartilage formation. Also, I evaluated the growth rate of chondrocyte cell line to measure the cellular proliferative potency. I have established the human auricular chondrocyte cell line integrated v-myc and confirmed by v-myc transduced Myc protein expression by immunohistochemistry and immunoblotting study. And, growth rate of established human auricular chondrocyte cell line increased 4 folds times faster than primarily cultured human auricular chondrocyte. The established human auricular chondrocyte had type II collagen mRNA upto 8 months in monolayer culture. And we observed formation of neocartilage on the back of nude mice using chondrocyte cell line/fibrin glue polymer at 12 weeks transplantation.
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Adult , Animals , Humans , Mice , Adhesives , Cell Line , Chondrocytes , Collagen Type II , Immunoblotting , Immunohistochemistry , Mice, Nude , Polymers , Retroviridae , RNA, MessengerABSTRACT
AIM: To investigate the effect of chicken collagen type II (C II) on collagen-induced arthritis (CIA) mice. METHODS: The model of mice CIA was prepared and the change of the mean of arthritis scores was observed. Meanwhile, ConA and LPS-induced, splenocytes proliferation and the production of interleukin (IL)-1 and IL-2 were measured. RESULT: CII [5, 10, 20 μg·kg-1ig × 7 d] significantly decreased the mean of arthritis scores, inhibited the increased ConA-induced splenocytes proliferation as well as the elevated production of IL-1 and IL- 2. Meanwhile, the pathological examination showed that articulus cartilage degeneration with synovial hyperplasia and inflammatory cells infiltration in CIA mice was suppressed by ig C II . CONCLUSION: C II has therapeutical effect on mice CIA which may be related to its immunoregulative function.
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Skeletogenesis occurs through either intramembranous or endochondral ossification. In addition, some parts of the skeletal components maintain their cartilaginous characteristics throughout life without mineralization. Runx2 is known to be a pivotal transcription factor for all skeletogenic processes. In this study, we examined the expression patterns of two major isoforms of Runx2 in early skeletogenesis. During intramembranous bone formation, Runx2-type I (Runx2-I) was widely expressed in osteoprogenitor cells and active osteoblasts, while Runx2-type II (Runx2-II) expression was stringently restricted to cells lining mineralized bones. Cells in permanent cartilage expressed collagen type II (Col-II) but never expressed Runx2 or Col-X. These permanent cartilages were well circumscribed by Runx2-I positive cells, in which Runx2-II was negative. In endochondral bone formation, Runx2 expression temporarily disappeared in Col-II-positive proliferating chondrocytes, but a secondary surge of Runx2-I expression occurred in the prehypertrophic zone before the mineralization of cartilage. Collectively, both Runx2 isoforms showed very similar expression patterns in active bone forming areas; however, Runx2-I has an exclusive role in the early commitment stage of intramembranous or endochondral bone forming processes or in cells surrounding permanent cartilage.