共查询到20条相似文献,搜索用时 10 毫秒
1.
《Journal of orthopaedic research》2017,35(4):829-836
2.
Melissa S. Ashwell Michael G. Gonda Kent Gray Christian Maltecca Audrey T. O'Nan Joseph P. Cassady Peter L. Mente 《Journal of orthopaedic research》2013,31(3):385-391
Our objective was to monitor chondrocyte gene expression at 0, 3, 7, and 14 days following in vitro impaction to the articular surface of porcine patellae. Patellar facets were either axially impacted with a cylindrical impactor (25 mm/s loading rate) to a load level of 2,000 N or not impacted to serve as controls. After being placed in organ culture for 0, 3, 7, or 14 days, total RNA was isolated from full thickness cartilage slices and gene expression measured for 17 genes by quantitative real‐time RT‐PCR. Targeted genes included those encoding proteins involved with biological stress, inflammation, or anabolism and catabolism of cartilage extracellular matrix. Some gene expression changes were detected on the day of impaction, but most significant changes occurred at 14 days in culture. At 14 days in culture, 10 of the 17 genes were differentially expressed with col1a1 most significantly up‐regulated in the impacted samples, suggesting impacted chondrocytes may have reverted to a fibroblast‐like phenotype. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 385–391, 2013 相似文献
3.
4.
[目的]研究降钙素(calcitonin, CT)对骨性关节炎关节软骨退变和软骨下骨骨代谢的影响.[方法]30只3个月龄雌性日本大耳白兔随机分为三组,其中两组行右膝关节前交叉韧带切断术(anterior cruciate ligament transaction,ACLT),分为ACLT+CT组和ACLT+NS组,第3组为Sham组.ACLT+CT给予每日1次皮下注射降钙素5 IU/(kg·d),持续8周,ACLT+NS组给予同样剂量生理盐水.术后8周后处死所有动物.取股骨髁制成切片行MMP-13和Ⅱ型胶原免疫组化染色.取胫骨近端制成硬组织切片行骨形态计量学检测.体外实验中,取兔膝关节软骨,经消化、培养,将第3代软骨细胞分三组:向IL-1β组加入人重组IL-1β(10 ng/ml). IL-1β+CT组加入人重组IL-1β (10 ng/ml)2 d后,再向培养液中加入CT(50 ng/ml).正常组不加任何诱导剂和干扰剂培养.然后行MMP-13、Ⅱ型胶原免疫组化检测和Realtime RT-PCR法检测.[结果]Sham组和ACLT+CT组软骨下骨骨小梁相对体积和厚度等均显著高于ACLT+NS组.Sham组和ACLT+CT组的Ⅱ型胶原的光密度值均显著高于ACLT+NS组,而MMP-13的光密度值显著低于ACLT+NS组(P<0.05).正常组和IL-1β+CT组的Ⅱ型胶原光密度值均显著高于IL-1β组而MMP-13的光密度值都显著低于IL-1β组(P<0.05).在正常组和IL-1β+CT组中Ⅱ型胶原的mRNA含量均显著高于IL-1β组而MMP-13的mRNA含量均显著低于IL-1β组(P<0.05).[结论]降钙素5 IU/(kg·d)皮下注射能够增加ACLT兔膝关节软骨Ⅱ型胶原的分泌和抑制MMP-13的表达,并可能通过调节软骨下骨的骨代谢和微结构来保护关节软骨; CT(50 ng/ml)能增加体外培养的含有IL-1β(10 ng/ml)的软骨细胞中Ⅱ型胶原的含量和抑制MMP-13分泌. 相似文献
5.
Alterations in structural macromolecules and chondrocyte deformations in lapine retropatellar cartilage 9 weeks after anterior cruciate ligament transection 下载免费PDF全文
Sang‐Kuy Han Ari P. Ronkainen Simo Saarakkala Lassi Rieppo Walter Herzog Rami K Korhonen 《Journal of orthopaedic research》2018,36(1):342-350
The structural integrity and mechanical environment of the articular cartilage matrix directly affect chondrocyte deformations. Rabbit models of early osteoarthritis at 9 weeks following anterior cruciate ligament transection (ACLT) have been shown to alter the deformation behavior of superficial zone chondrocytes in mechanically loaded articular cartilage. However, it is not fully understood whether these changes in cell mechanics are caused by changes in structural macromolecules in the extracellular matrix. Therefore, the purpose of this study was to characterize the proteoglycan content, collagen content, and collagen orientation at 9 weeks post ACLT using microscopic techniques, and relate these changes to the altered cell mechanics observed upon mechanical loading of cartilage. At 9 weeks following ACLT, collagen orientation was significantly (p < 0.05) altered and proteoglycan content was significantly (p < 0.05) reduced in the superficial zone cartilage matrix. These structural changes either in the extracellular or pericellular matrix (ECM and PCM) were also correlated significantly (p < 0.05) with chondrocyte width and height changes, thereby suggesting that chondrocyte deformation response to mechanical compression in early OA changes primarily because of alterations in matrix structure. However, compared to the normal group, proteoglycan content in the PCM from the ACLT group decreased less than that in the surrounding ECM. Therefore, PCM could play a key role to protect excessive chondrocyte deformations in the ACLT group. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:342–350, 2018. 相似文献
6.
目的观察膝关节原发性骨关节炎(osteoarthritis,OA)胫骨平台软骨和软骨下骨病理改变特点,对比内、外侧平台软骨和软骨下骨结构参数,探讨钙化层和软骨下骨在OA发病机制中的作用。方法取2009年10月-2011年5月行人工全膝关节置换术治疗的30例30膝原发性OA患者自愿捐赠的新鲜胫骨平台标本进行实验。其中男11例,女19例;年龄55~78岁,平均65.1岁。病程10~25年,平均16.6年;患膝内翻畸形1~23°,平均9.3°。大体观察胫骨平台后在内、外侧中央负重区取材,常规制备脱钙石蜡切片,行HE和番红O/固绿染色,观察关节软骨退变特点,参照Mankin评分标准评分并分期;观察钙化层及软骨下骨病理改变。应用Image Pro Plus 6.0图像分析软件测量软骨和软骨下骨结构参数,包括软骨全层(total articular cartilage,TAC)厚度、钙化层(articular calcified cartilage,ACC)厚度、ACC/TAC比值、软骨下骨板(subchondral bone plate,SCP)厚度以及骨小梁体积分数(trabecular bone volume,BV/TV)。结果大体观察内侧平台软骨退变较外侧严重,内侧平台软骨Mankin评分为(12.4±1.1)分,显著高于外侧平台的(8.3±1.6)分(t=12.173,P=0.000)。根据Mankin评分结果在60个标本中,14个为OA早期,可见软骨浅表层裂隙、潮线复制和软骨下骨增厚;19个为OA中期,可见软骨深层裂隙、多发软骨下骨吸收陷窝和明显增厚的软骨下骨;27个为OA晚期,可见软骨全层缺失、软骨内化骨和"象牙化"软骨下骨。软骨和软骨下骨结构参数测定示:内侧平台TAC厚度显著低于外侧平台,ACC/TAC比值、BV/TV及SCP厚度显著高于外侧平台,差异均有统计学意义(P<0.05)。内、外侧平台ACC厚度比较,差异无统计学意义(P>0.05)。结论钙化层和软骨下骨可能在OA发生与进展中发挥了重要作用。 相似文献
7.
Dong Zhang Lanny J Johnson Hu-Ping Hsu Myron Spector 《Journal of orthopaedic research》2007,25(7):873-883
The objective of this study was to identify and characterize cartilaginous deposits aggregates in the subchondral bone in areas of the human osteoarthritic knee with exposed bone. A specific aim was to determine the distribution of the joint lubrication molecule, lubricin/superficial zone protein [referred to by its gene, proteoglycan4 (PRG4)], in these cartilaginous deposits and in osteoarthritic cartilage. This work was carried out in the context of assessing the potential contribution of these chondrocyte aggregates for joint resurfacing in certain cartilage repair procedures. The discarded bone cuts of femoral condyles and tibial plateaus were collected from 11 patients with advanced osteoarthritis (OA) of the knee during total knee arthroplasty; 9 women and 2 men with a mean age of 68 years. Sections of paraffin-embedded tissue were stained with Safranin-O, and with antibodies to type II collagen, alpha-smooth muscle actin (SMA), and PRG4. Chondrocyte aggregates were found in the subchondral bone of regions of exposed bone in sections from five individuals. The average diameter of cartilaginous aggregates was 152 microm, and the average depth of the aggregates below the surface was about 475 microm. Most aggregates were fibrocartilaginous and stained positive for type II collagen. Of interest was the finding that the cartilaginous deposits and osteoarthritic cartilage contained PRG4. Only a small percentage of chondrocytes stained positive for SMA. Cartilaginous deposits containing chondrocyte aggregates exist in subchondral bone in regions of exposed bone in some patients with advanced OA of the knee. These cells may be able to contribute to the resurfacing of the joint in certain cartilage repair procedures. 相似文献
8.
《Journal of orthopaedic research》2017,35(4):858-867
9.
The effect of insulin-dependent diabetes mellitus (IDDM) on bone metabolism was evaluated using the streptozotocin (STZ)-induced diabetic rat 1 week after the induction of diabetes. The urinary excretion of cross-linked N-telopeptides of type I collagen (NTx) and deoxypyridinoline (Dpd) in diabetic rats increased to 3.6-fold and 1.2-fold the control level, respectively. The amount of hydroxyproline and calcium in the distal femur of diabetic rats significantly decreased to 76% and 90% of the control, respectively. The levels of serum osteocalcin and alkaline phosphatase (ALP) activity in the distal femur of the diabetic rats were significantly reduced to about 40% and 70% of the control levels, respectively. The decrease in the expression osteocalcin was observed in distal femur of the diabetic rats, although the level of ALP mRNA was unchanged. The activity and the mRNA level of tartrate-resistant acid phosphatase (TRAP) increased to 1.5- and 2.3-fold the control level, respectively, in distal femur of the diabetic rats. The activity, protein, and mRNA levels of cathepsin K of diabetic rats also elevated to about 2-, 2.3-, and 2-fold the control levels, respectively. These results suggest that IDDM contributes to bone loss through changes in gene expression of TRAP and cathepsin K in osteoclasts as well as osteocalcin in osteoblasts resulting in increased bone resorptive activity and decreased bone formation. 相似文献
10.
Noboru Yatsugi Tomoo Tsukazaki Makoto Osaki Takehiko Koji Shunichi Yamashita Hiroyuku Shindo 《Journal of orthopaedic science》2000,5(2):150-156
To investigate the relationship of chondrocyte apoptosis and cartilage destruction, we performed in situ nick end labeling
(ISNEL), electron microscopy, and im-munohistochemistry against apoptosis-related proteins, p53 and c-myc, in the articular
cartilages of patients with rheumatoid arthritis (RA; n = 12) and osteoarthritis (OA; n = 12), and in control articular cartilages from patients with femoral neck fracture (n = 8). The distribution of stained chondrocytes was evaluated semiquantitatively in relation to the degree of cartilage destruction.
ISNEL-positive chondrocytes with apoptotic morphological features were identified in a relatively early phase of cartilage
destruction, and correlated positively and significantly in a number with the degree of cartilage degeneration. Comparison
of RA and OA revealed a significantly greater number of ISNEL-positive chondrocytes in RA cartilage. In contrast, the specimens
of normal subjects contained few cells with apoptotic changes. Similarly to the distribution of ISNEL staining, the expression
of p53 and c-myc proteins was observed in chondrocytes within the degraded lesions, and showed a positive correlation with
the number of ISNEL-stained cells. These results suggest that the degree of chondrocyte apoptosis is closely related to cartilage
destruction and that chondrocytes in RA more readily undergo apoptosis than those in OA. The expression of p53 and c-myc proteins
in ISNEL-positive areas may reflect the involvement of these proteins in the apoptotic process in articular chondrocytes in
inflammatory arthritis.
Received for publication on April 28, 1999; accepted on Sept. 22, 1999 相似文献
11.
12.
[目的]用TUNEL标记法和透射电镜来观察软骨撞击伤后软骨细胞凋亡的演变情况。[方法]56只健康成年新西兰大白兔,随机分高、低能量两组,每组28只,随机选取一侧后肢为实验组,另一侧为对照组;不同重量的击锤撞击股骨内侧髁软骨面致不同的损伤。术后4 d,1、2、4、8、16、32周收集标本,每个时间组4只兔子,进行TUNEL标记法和透射电镜来观察软骨细胞的凋亡。[结果]对照组标本中在软骨的中间层和钙化层,出现凋亡细胞。低能量组中,伤后4 d,1、2周的标本中,软骨的移形层和过渡层出现凋亡细胞,细胞凋亡率与对照组之间无统计学意义;4周后的标本中凋亡细胞明显减少,细胞凋亡率与对照组之间无统计学意义。高能量组中,伤后4 d软骨浅表层和移形层中出现凋亡细胞,而且伤后时间越长凋亡细胞所占比率越大,细胞凋亡率与对照组之间有统计学意义。[结论]软骨高、低能量撞击伤后早期软骨细胞均出现凋亡,凋亡率高于对照组;但低能量伤后4周软骨细胞凋亡率恢复到对照组水平;高能量伤后软骨细胞凋亡率随时间逐渐增高。 相似文献
13.
Fibronectin splice variation in human knee cartilage,meniscus and synovial membrane: Observations in osteoarthritic knee 下载免费PDF全文
Carla R. Scanzello Dessislava Z. Markova Ana Chee Yan Xiu Sherrill L. Adams Greg Anderson Miltiadis Zgonis Ling Qin Howard S. An Yejia Zhang 《Journal of orthopaedic research》2015,33(4):556-562
14.
15.
Tiffany Cheng Nicole C. Maddox Andrew W. Wong Ruyan Rahnama Alfred C. Kuo 《Journal of orthopaedic research》2012,30(2):234-245
During monolayer culture, articular chondrocytes dedifferentiate into fibroblast‐like cells. The mechanisms underlying this process are poorly understood. We sought to further characterize dedifferentiation by identifying an extended panel of genes that distinguish articular cartilage from dedifferentiated chondrocytes. Thirty‐nine candidate marker‐genes were identified from previous studies on articular‐cartilage gene‐expression. Real‐time PCR was used to evaluate the mRNA levels for these candidates in calf articular cartilage and dedifferentiated articular chondrocytes. Twenty‐two of the candidate marker genes exhibited at least a two‐fold difference in gene expression in the two cell types. Twelve of these genes had at least a ten‐fold difference in gene expression. Tenascin C (TNC), type I collagen (COL1A1), and hypoxia‐inducible factor 1 alpha (HIF1α) showed the highest relative expression levels in dedifferentiated chonodrocytes. Type II collagen (COL2A1), type XI collagen (COL11A2), and superficial zone protein (SZP) showed the highest relative expression levels in articular cartilage. In contrast to previous findings, fibromodulin mRNA, and protein levels were higher in dedifferentiated chondrocytes. Compared to smaller subsets of markers, this panel of 12 highly differentially expressed genes may more precisely distinguish articular cartilage from dedifferentiated chondrocytes. Since many of the genes up‐regulated in dedifferentiated chondrocytes are also expressed during cartilage development, dedifferentiated chondrocytes may possess features of cartilage precursor cells. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 30:234–245, 2012 相似文献
16.
Subchondral bone fragility with meniscal tear accelerates and parathyroid hormone decelerates articular cartilage degeneration in rat osteoarthritis model 下载免费PDF全文
Yugo Morita Hiromu Ito Masahiro Ishikawa Takayuki Fujii Moritoshi Furu Masayuki Azukizawa Akinori Okahata Takuya Tomizawa Shinichi Kuriyama Shinichiro Nakamura Kohei Nishitani Hiroyuki Yoshitomi Shuichi Matsuda 《Journal of orthopaedic research》2018,36(7):1959-1968
17.
Yan Lu Mark D Markel Carol Swain Lee D Kaplan 《Journal of orthopaedic research》2006,24(10):1974-1982
The purpose of this study was to create a controlled partial thickness cartilage lesion in a sheep model, and to provide a foundation to study the natural history of the progression of this lesion. Twenty-eight sheep divided into four groups (1, 12, 24, and 52 weeks, n=7/group) were used in this study. In one stifle, a mechanical tool was used to create a 200 microm partial thickness lesion (1.5x1.5 cm2) on the medial femoral condyle via arthroscopy. Joint fluid was drawn presurgery and after euthanasia for analysis of collage II 3/4 C (long) (C2C). After euthanasia, the condyle was analyzed by gross appearance, confocal laser microscopy (CLM) for cell viability, scanning electronic microscopy (SEM) for surface roughness, Artscan for cartilage stiffness, and histology for cartilage morphology. The gross appearance of the treated area appeared rough, soft, and swollen compared to untreated control over time. CLM demonstrated that the depth of cell death increased to 590 microm at 52 weeks after surgery. SEM demonstrated that the treated area became more irregular over time. Stiffness of the treated area was significantly less than control by 12 weeks after surgery. Histologic analysis demonstrated that the 12, 24, and 52 week groups had significantly poorer histologic scores than the 1 week group. Joint fluid analysis demonstrated that the treatment group at 1 week had significant higher levels of C2C than the pretreatment baseline data. The results of this study demonstrated that partial thickness injury of cartilage continued to propagate and degenerate over time in this sheep model. Options for the prevention or treatment of this lesion may be tested using this model in the future. 相似文献
18.
人负重关节软骨细胞的体外分离与培养 总被引:2,自引:0,他引:2
目的 研究人负重关节软骨细胞的分离培养技术。方法 对 10例手术切除的负重关节软骨进行0 2 %Ⅱ型胶原酶分离、10 %DMEM中培养 ,观察软骨细胞获得率、贴壁率、增殖状况及形态学改变。结果 ① 0 2 %Ⅱ型胶原酶消化尽 2 0 0~ 70 0mg软骨标本需 4~ 14h ,细胞获得率 (1 82± 0 5 7)× 10 6/g ,存活率(79 4± 9 9) %。②细胞可传 6~ 12代 ,前 4代培养时间平均 (33± 10 )d ,生长倍数平均 198倍 ,倍增时间(4 13± 1 34)d。③第 3、4代细胞呈多角形、部分梭形 ,8、9代后细胞梭状铺平。结论 30 0~ 5 0 0mg软骨标本 ,经该方法培养 3~ 4周后可达第 4代 ,细胞数可达 (0 6~ 1 0 )× 10 8个 ,可满足临床上修复 15~ 2 5cm2软骨缺损的需要。 相似文献
19.
Simon Y. Tang Richard B. Souza Michael Ries Paul K. Hansma Tamara Alliston Xiaojuan Li 《Journal of orthopaedic research》2011,29(9):1312-1319
The objective of this study is to examine the local relationship between T1ρ relaxation times and the mechanical behavior of human osteoarthritic articular cartilage using high‐resolution magnetic resonance imaging (MRI) and local in situ microindentation. Seven human tibial plateaus were obtained from patients who underwent total knee arthroplasty due to severe osteoarthritis (OA). Three to six sites were selected from each sample for visual classification using the ICRS Outerbridge scale (a total of 36 sites). Samples were imaged by MR, and the local distribution of T1ρ relaxation times were obtained at these selected sites. The elastic and viscoelastic characteristics of the tissue were quantified nondestructively using dynamic microindentation to measure peak dynamic modulus, energy dissipation, and phase angle. Measured Outerbridge scores, MR T1ρ relaxation times, and mechanical properties were highly heterogeneous across each cartilage surface. Site‐specific measures of T1ρ relaxation times correlated significantly with the phase angle (p < 0.001; R = 0.908), a viscoelastic mechanical behavior of the cartilage. The novel combination of high‐resolution MR imaging and microindentation allows the investigation of the local relationship between quantitative MRI and biomechanical properties in highly heterogeneous OA cartilage. These findings suggest that MRI T1ρ can provide a functional assessment of articular cartilage. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1312–1319, 2011 相似文献
20.
The labeling of the articular surface with cationized ferritin (CF), an electron–dense marker, visualizes the anionic sites and may disclose abnormal penetration of the large CF molecule into the subsurface layers. Various areas of cartilage selected by unaided eye examination were taken from femoral heads excised in three cases of osteoarthritis and two cases of hip fracture. The fragments were examined by optical microscopy and by electron microscopy after labeling with CF. The labeling with and the penetration of CF were correlated with the morphological features of the surface. The surfaces belonging to the erosion border were disrupted and the CF penetrated approximately 2 μm into the matrix along the collagen fibers and in areas containing a patchy dense material. Prefixation with Karnovsky's fixative prevents CF penetration. The fragments taken at a distance from the erosion border showed at electron microscopical examination either an intact appearance of the surface that was labeled without penetration or a disrupted surface with penetration of the label. The osteophytes and the regeneration buds surface were labeled showing little or no penetration. The fragments from cartilage of hip fractures had either an intact surface regularly labeled or a slightly or moderately disrupted surface with moderate penetration of CF. The penetration of large molecules of CF in damaged cartilage demonstrates important permeability changes that may be significant for the pathogenetic mechanism of osteoarthritis. Similar permeability changes were previously shown in mice femoral heads treated in vitro with collagenase or trypsin and labeled with CF. 相似文献