Four children having the main features of limp, pain in the hip, limitation of motion and external rotation of the affected limb going through MRI assessment, surgical exploration of the affected hip and the responses to various methods of treatment. It was found that the impingement of synovium in between the femoral head and the acetabulum was the chief pathology. The nomenclature, classification and clinical importance, pathogenesis and the differential diagnosis were diseussed. This specific group of patients were given under the nomenclature as specific type of transient synovitis of hip in children-intraaticular synovial impingement type.
To study the method of isolating and culturing synovium-derived MSCs (SMSCs), and to investigate its multiple differentiation potential in vitro. Methods Three 2-month-old Changfeng hybrid swines weighing 8-10 kg (male and female) were used. SMSCs were harvested from the synovium of swine knee joints and cultured in vitro. When the SMSCs at passage 3 reached confluence, basic culture medium was removed, and the multi ple differentiationpotential of SMSCs was demonstrated in specific induction media (experimental group). The cells at passage 3 cultured with basic culture medium served as control group. After 21 days of chondrogenic differentiation, the cells underwent toluidine blue staining, immunohistochemistry staining and real-time fluorescence quantitative PCR detection. After 10 and 21 days of osteogenic differentiation, the cells underwent ALP staining and Al izarin red staining, respectively. After 21 days of adipogenic differentiation, the cells underwent Oil red O staining. Results SMSCs displayed long and thin or polygonal morphology 24 hours after culture. They prol iferated fast 48 hours after culture and presented large number of spindle-shaped cells with few globular cells 72 hours after culture. For the experimental group 21 days after chondrogenic induction, the cells were positive for toluidine blue staining with the formation of Aggrecan outside the cells; the immunohistochemistry staining revealed the expression of Col II; the real-time fluorescence quantitative PCR detection showed that the expressions of Col II A1, Aggrecan and SOX9 mRNA of the experimental group were greater than that of control group (P lt; 0.05). The cells were positive for ALP staining 10 days after osteogenic induction, and positive for Al izarin red staining 21 days after osteogenic induction, with the formation of calcium nodules. Oil red O staining displayed the formation of l i pid droplets inside the cells 21 days after adi pogenic induction. For the control group, the results of all the staining assays were negative except the ALP staining presenting with sl ight positive result. Conclusion SMSCs can be isolated from knee joint of swine and proliferate and differentiate into osteogenic, adi pogenic and chondrogenic cells in vitro. SMSCs may be a promising source of seed cells for tissue engineering.
ObjectiveTo explore the pathological role of high mobility group box chromosomal protein 1 (HMGB1) in osteoarthritis (OA) by comparing the difference of HMGB1 in the synoviocytes between OA and normal knees. MethodsSynoviocyte lines from OA and normal knees were collected and cultured. Immunohistochemistry and Western blot were applied to identify the difference of HMGB1 between the OA and normal synoviocyte lines. The eukaryotic expression vector containing human Pgenesil-1/HMGB1 small interfering RNA (siRNA) were constructed and identified. The synoviocyte lines were transfected with the eukaryotic expression vector of Pgenesil-1/HMGB1 siRNA (Pgenesil-1/HMGB1 siRNA group) and with Pgenesil-1 plasmid (Pgenesil-1 group) and were not transfected as a control (untransfected group). Western blot was applied to identify the difference of HMGB1 among groups, and the levels of interleukin 1β (IL-1β) and tumor necrosis factor α (TNF-α) protein synthesis in the supernatants were measured by ELISA. ResultsPrimary knee synoviocytes cultured in vitro were fibroblast-like cells with longspindle shape. The immunohistochemistry and immunofluorescence results showed positive staining for HMGB1 in cytoplasm and weak positive staining in the nucleus in the OA synoviocyte line, but positive staining for HMGB1 in the nucleus and weak positive staining in the cytoplasm in the synoviocyte line of normal knee. The level of HMGB1 in the OA synoviocytes (0.687±0.025) was significantly higher than that of normal synoviocytes (0.172±0.030) (t=32.159, P=0.000) by Western blot. The recombinant plasmid Pgenesil-1/HMGB1 siRNA was successfully constructed. The expression of HMGB1 protein in Pgenesil-1/HMGB1 siRNA group (0.134±0.048) was significantly lower than that of Pgenesil-1 group (0.581±0.032) and untransfected group (0.514±0.069) (P<0.05). ELISA results showed that IL-1β and TNF-α in supernatants of Pgenesil-1/HMGB1 siRNA group were significantly lower than those of Pgenesil-1 group and untransfected group (P<0.05). ConclusionThe up-regulated expression and expressed location (from nucleus to cytoplasm) of HMGB1 in the synoviocyte are closely related to OA. The siRNA targeting inhibition of HMGB1 gene expression can obviously inhibit IL-1β and TNF-α in supernatants of the OA synoviocyte line and delayed the inflammation of OA.
Objective To explore the method and outcome of knee resurfacing arthroplasty in treating late-staged diffuse pigmented villonodular synovitis (PVNS). Methods Between November 2002 and May 2009, 11 cases of late-staged diffuse PVNS were treated, including 3 males and 8 females with an average age of 51.2 years (range, 42-63 years). The diseaseduration was 2.5-10.0 years (mean, 5.2 years). Unilateral knee was involved in all patients, including 7 left knees and 4 right knees. Nine patients had a history of trauma and 2 cases had no obvious inducing factors. The range of motion was (90.1 ± 17.2)° and Hospital for Special Surgery Knee Score (HSS) was 68.9 ± 8.7. After synovectomy, knee resurfacing arthroplasty was performed in all patients. Results Superficial infection of the incision occurred in 1 case at 6 days postoperatively and was cured after debridement; other incisions healed by first intention. Limited flexion and extension, incomplete palsy of common peroneal nerve, and deep venous thrombosis occurred in 1 case respectively, and were cured or improved after symptomatic treatment. All the 11 cases were followed up 38 months on median (range, 13 to 102 months). Two cases developed chronic pain and were not given treatment. Recurrence occurred in 1 case 12 months postoperatively and recovered after synovectomy again. X-ray films showed no signs of loosening, sinking, and bone destruction. At last follow-up, the range of motion was (109.1 ± 18.6)° and HSS score was 86.7 ± 9.3, showing significant differences when compared with those before operation (P lt; 0.05). According to the HSS score system, the results were excellent in 6 cases, good in 3, fair in 1, bad in 1, and the excellent and good rate was 81.8%. Conclusion A combination of knee resurfacing arthroplasty and synovectomy for the treatment of late-staged diffuse PVNS is able to get a good cl inical results in restoration of function, improvememt of the l ife quality, and decrease of recurrence rate.
ObjectiveTo investigate the anti-apoptotic ability of synovium-derived mesenchymal stem cells (SMSCs) by comparing the apoptosis induced by tumor necrosis factor α (TNF-α) between SMSCs and bone marrow mesenchymal stem cells (BMSCs). MethodSMSCs and BMSCs were isolated with tissue adhering and density gradient centrifugation respectively, and cells at passages 3-5 were used in further experiments. After immunophenotype identification and differentiation induction, cells were divided into 4 groups. In the experimental groups, apoptosis of SMSCs and BMSCs were induced by 20 ng/mL TNF-α and 10 μg/mL cycloheximide, and cells were cultured in normal culture medium in the control groups. Cellular morphology were observed by inverted phase contrast microscope. After apoptosis induction for 24 hours, cell viability was determined by cell counting kit 8 assay and apoptotic index was detected by flow cytometer. Moreover, the level of Cleaved Caspase-8, 3 were determined by Western blot. ResultsBoth SMSCs and BMSCs accorded with the definition criteria of MSCs according to results of immunophenotype identification and differentiation induction. After apoptosis induction, cells became shrinking and partially floated and cellular morphologies became worse than those in the control groups. After apoptosis induction for 24 hours, cell viabilities of SMSCs and BMSCs in the control groups were both 100%, and no apoptotic cells were observed. However, cell viabilities of SMSCs and BMSCs in the experimental groups were 60.13%±8.63% and 46.55%±10.54% respectively, which were both significantly lower than those in the control groups (P<0.05) , and cell viability in the SMSCs experimental group was significantly higher than that in the BMSCs experimental group (t=3.152, P=0.006) . The apoptotic index was 36.54%±8.63% in the SMSCs experimental group and was 53.77%±11.52% in the BMSCs experimental group, both were significantly higher than the control groups (1.12%±0.24% and 1.35%±0.31%) (P<0.05) . What's more, it was significantly lower in SMSCs experimental group than that in BMSCs experimental group (t=3.785, P=0.001) . Moreover, no expression of Cleaved Caspase-8, 3 was detected in the control groups. But the levels of Cleaved Caspase-8, 3 were significantly enhanced in the experimental groups and they were lower in SMSCs than in BMSCs (t=13.870, P=0.000; t=7.309, P=0.000) . ConclusionsTNF-α induced apoptosis is lower in SMSCs than in BMSCs, which means that SMSCs may have stronger anti-apoptosis ability than BMSCs.
Objective To investigate the method and the effectiveness of arthroscopy and/or arthrotomy combinedwith postoperative radiotherapy for diffuse pigmented villonodular synovitis (PVNS) of the knee. Methods BetweenSeptember 2000 and August 2010, 97 patients with diffuse PVNS of the knee were treated. There were 38 males and 59 femaleswith a median age of 33 years (range, 8-75 years). The disease duration ranged from 1 week to 30 years, including 52 left kneesand 45 right knees. There were 10 recurrent cases. The extention and flexion of the knee joint were (1.9 ± 2.3)° and (122.9 ± 5.6)°,respectively; the Lysholm score was 43.2 ± 6.7; and the International Knee Documentation Committee (IKDC) score was53.2 ± 5.7, preoperatively. According to the scope and degree of the knee joint lesions, simultaneous anterior and posteriorsynovectomy was performed under arthroscopy in 82 cases, synovectomy under arthroscopy and removal of posterior extraarticularlesion by arthrotomy in 3 cases, synovectomy and the soft tissue lesions resection under arthroscopy in 9 cases, andstaging resection and bone graft in 3 cases. After operation, 76 patients received postoperative radiotherapy. Results Poplitealartery was injuryed in 1 case and the branch of popl iteal veins were injuryed in 3 cases during operation. Intra-articularhemorrhage occurred in 1 case at 3 days after operation. The other patients achieved heal ing of incision by first intentionwithout nerve damage and other complications. All patients were followed up 1 year and 3 months to 11 years and 2 months(median, 61 months) postoperatively. During follow-up, 89 cases had no relapse. At 15 months after operation, the extentionand flexion of the knee joint were (0.2 ± 1.3)° and (135.9 ± 6.6)°, respectively; the Lysholm score was 89.8 ± 5.8; and the IKDCscore was 87.8 ± 5.8. All indexes were significantly improved when compared with the preoperative ones (P lt; 0.05). At 6 monthsto 8 years postoperatively, 8 cases had occurrence, and they had sl ight limitation of the range of motion but had no pain andswelling of the knees after reoperation. Conclusion According to the scope and degree of the knee joint lesions, arthroscopyand/or arthrotomy combined with postoperative radiotherapy should be chosen for diffuse PVNS of the knee so as to obtain good effectiveness. Radiotherapy and enough total radiation dose are important factors to insure no recurrence.
Objective To investigate the feasibility of rabbit synovial-derived mesenchymal stem cells (SMSCs) differentiating into fibrocartilage cells by the recombinant adenovirus vector mediated by bone morphogenetic protein 2/7 (BMP-2/7) genes in vitro. Methods SMSCs were isolated and purified from 3-month-old New Zealand white rabbits [male or female, weighing (2.1 ± 0.3) kg]; the morphology was observed; the cells were identified with immunocytological fluorescent staining, flow cytometry, and cell cycles. The adipogenic, osteogenic, and chondrogenic differentiations were detected. The recombinant plasmid of pAdTrack-BMP-2-internal ribosome entry site (IRES)-BMP-7 was constructed and then was used to infect SMSCs. The cell DNA content and the oncogenicity were tested to determine the safety. Then infected SMSCs were cultured in incomplete chondrogenic medium in vitro. Chondrogenic differentiation of infected SMSCs was detected by RT-PCR, immunofluorescent staining, and toluidine blue staining. Results SMSCs expressed surface markers of stem cells, and had multi-directional potential. The transfection efficiency of SMSCs infected by recombinant plasmid of pAdTrack-BMP-2-IRES-BMP-7 was about 70%. The safety results showed that infected SMSCs had normal double time, normal chromosome number, and normal DNA content and had no oncogenicity. At 21 days after cultured in incomplete chondrocyte medium, RT-PCR results showed SMSCs had increased expressions of collegan type I and collegan type II, particularly collegan type II; the expressions of RhoA and Sox-9 increased obviously. Immunofluorescent staining and toluidine blue staining showed differentiation of SMSCs into fibrocartilage cells. Conclusion It is safe to use pAdTrack-BMP-2-IRES-BMP-7 for infecting SMSCs. SMSCs infected by pAdTrack-BMP-2-IRES-BMP-7 can differentiate into fibrocartilage cells spontaneously in vitro.
ObjectiveTo investigate the role of CXCL13 in the onset and development of knee osteoarthritis by observing and comparing the expression of CXCL13 between osteoarthritis and normal synovium. MethodsThe synovium samples were collected from 30 patients with osteoarthritis who received total knee replacement (osteoarthritis group), including 11 males and 19 females with an average age of 66.7 years (range, 62-76 years). The synovium samples were collected from 22 patients without osteoarthritis who underwent traumatic amputation (control group), including 15 males and 7 females with an average age of 51.3 years (range, 48-56 years). The NimbleGen microarray detection was used to defect differentially expressed genes; the immunohistochemistry staining, Western blot, and real-time quantitative PCR (qRT-PCR) were used to detect the expressions of CXCL13 mRNA and protein. ResultsThere were 451 up-regulated genes and 810 down-regulated genes in the 22 885 genes which contained by mRNA gene chip, and CXCL13 gene expression was down-regulated. Immunohistochemistry staining and Western blot assay showed that the expression of CXCL13 protein was significantly lower in osteoarthritis group (0.408 0±0.101 8) than in control group (0.785 9±0.057 9) (t=15.630, P=0.000). qRT-PCR results showed that the expression of CXCL13 mRNA was significantly lower in osteoarthritis group (0.011 7±0.003 2) than in control group (1.041 4±0.129 7) (t=43.634, P=0.000). ConclusionLow expression of CXCL13 in the knee osteoarthritis synovium tissue may be associated with the onset and development of knee osteoarthritis.
【摘要】 目的 探討關節鏡治療膝關節滑膜軟骨瘤病的療效。 方法 2005年1月—2009年10月，對23例（28膝）滑膜軟骨瘤病患者入院行X線片、關節活動度檢查、視覺模擬評分以及Lysholm膝關節功能評分。根據鏡下所見分為表淺型6例，游離體型17例。結合病理學檢查行Milgram 分期，Ⅱ期16例，Ⅲ期7例。所有患者均行關節鏡下病變滑膜切除及游離體取出治療。 結果 所有患者均隨訪13~57個月，平均（32.3±6.7）個月，術后傷口均甲級愈合。術后（5.05±2.43） d恢復正常生活或工作。癥狀明顯改善21例（91.30%），部分改善2例（8.70%），對療效滿意23例（100%）。膝關節關節活動度由術前的伸膝（14.29±16.34）°以及屈膝（106.07±35.83）°提高到術后的伸膝（1.79±2.79）°及屈膝（132.64±35.64）°，差異具有統計學意義（Plt;0.05）。負重行走時疼痛視覺模擬評分由術前的（3.81±2.02）分降低到術后的（0.37±0.65）分（Plt;0.05）。Lysholm評分由術前的（43.20±8.24）分升至術后6個月的（86.72±5.40）分（Plt;0.05）；術后1年復診并檢查膝關節正側位X線片，均未見滑膜軟骨瘤體，所有患者無復發。 結論 關節鏡下游離體取出術聯合病變滑膜切除術療效滿意，關節疼痛明顯減輕，功能恢復，是一種治療膝關節滑膜軟骨瘤病確切有效的方法。【Abstract】 Objective To investigate the therapeutic effect of arthroscopic treatment on synovial chondromatosis. Methods A total of 23 patients (28 knees) with synovial chondromatosis were diagnosed and treated in our hospital from January 2005 to October 2009. All of the patients underwent radiographic imaging examination, knee joint range of motion (ROM), visual analogue scale (VAS) and Lysholm score. According to distinct arthroscopic appearance, superficial pattern was found in 6 patients and loose body lesion pattern was in 17. Additionally, combined with pathological examination, according to the Milgram staging，Stage Ⅱ was in 6 patients and Stage Ⅲ was in 7. Arthroscopic limited synovectomy and removal of loose bodies were performed on all the patients. Results The patients were followed up for 13-57 months with the mean of (32.3±6.7) months. The wound of all patients healed up. The time of returning to normal work and life was (5.05±2.43) days for average. The postoperative symptom was markedly alleviated in 21 patietns and partly alleviated in 2. All patients were satisfied with the therapeutic effect. The mean activity of knee joint was significantly different befoe and after the surgery (Plt;0.05) preoperative extension and flexion degrees were (14.29±16.34) and (106.07±35.83) degrees, respectively; postoperative extension and flexion degrees were (1.79±2.79) and (132.64±35.64) degrees (flexion) , respectively. The mean VAS score of weight bearing walking was 0.37±0.65 after theoperation and 3.81±2.02 before the peration; the difference was significantly different (Plt;0.05). The preoperative Lysholm knee score was 34-67 with the mean of 43.20±8.24, and the post-operative score was 71-99 with the mean of 86.72±5.40. There were differences in preoperative and post-operative scores (Plt;0.05) . Radiographic imaging examination of knee joint was performed 1 year after the opertation, no loose bodies was seen and no patients recurred. Conclusion The therapeutic effect of arthroscopic limited synovectomy and removal of loose bodies is good on synovial chondromatosis.