From March. 1987 to March. 1989,we have treated 8cases of children with avascular necrosis of the femoral headby synovectomy of the hip and lateral circumflex femoralartery pedicled iliac bone graft to the femoral neck. Satisfac- tory therapeutic results were achived. The advantages of thisoperation are : 1. the microcirculation of the femoral headwas improved-by intraarticular decompression. 2. the venouspressure decreased by osteotomy at femoral head and neck.3. iliac bone graft can prevent femoral head coiiapsc.4.the blood supply of the femoral head was recstablished by vascularized iliac bone gredt.
Objective To study the effects of the periosteum，synovium andcartilage tissues on the gene expressions of proteoglycan, collagen Ⅱ, andnuclear factor kappa B (NF-κB) and to investigate the different effects of these tissues on cartilage regeneration. Methods In 20 New Zealand white rabbits, 20 cartilage explants were taken from the knee joints in each rabbit, the sizeof which was 4 mm×4 mm×4 mm. All the cartilages were divided into the following 4 groups and cultured for 7 days: Group A, with 5 pieces (2 mm×2 mm) of the synovium of theknee joints in each dish; Group B, with 5 pieces (2 mm×2 mm) of the periosteum ineach dish; Group C, with 5 pieces (2 mm×2 mm×2 mm) of the cartilage in each dish; and Group D, with no addition of other tissues (control group). RNA was extracted from the cells of the cartilage explants (4 mm×4 mm×4 mm) in all the dishes. Thegene expressions of proteoglycan, collagen Ⅱ and NF-κB were defected by a reversetranscription-polymerase chain reaction (RT-PCR).Results In group A, the gene expression of proteoglycan was significantly decreased. The relative density of this gene expression had a significant difference when compared with that in group D (1.09±0.21 vs. 1.25±0.25, Plt;0.05); the gene expressions of collagen Ⅱ and NF-κB were also decreased, but they had no significant differences when compared with those in group D (Pgt;0.05). In groupB, the gene expressions of proteoglycan, collagen Ⅱ, and NF-κB were significantly increased. The relative densities of these gene expressions were 1.60±0.26, 1.57±0.24, and 4.20±2.22, respectively, which had significant differences when compared with those in group D (Plt;0.05). In group C, the relative density of the gene expression of collagen Ⅱ was 1.43±0.28, which had a significant difference when compared with that in group D (Plt;0.05), but therelative densities of the gene expressions of proteoglycan and NF-κB had no significant differences when compared with those in group D (Pgt;0.05). Conclusion The results indicate that the periosteum can up-regulate the gene expressions of proteoglycan, collagen Ⅱ and NF-κB. The NF-κB is likely to be an important nuclear transcription factor related to cartilage regeneration. The results also suggest that the periosteum maybe better in facilitating the cartilage repair and regeneration in clinical practice.
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 explore the conditions of synovial derived mesenchymal stem cells (SMSCs) differentiating into the fibrocartilage cells by using the orthogonal experiment.
MethodsThe synovium was harvested from 5 adult New Zealand white rabbits, and SMSCs were separated by adherence method. The flow cytometry and multidirectional differentiation method were used to identify the SMSCs. The conditions were found from the preliminary experiment and literature review. The missing test was carried out to screen the conditions and then 12 conditions were used for the orthogonal experiment, including transforming growth factor β1 (TGF-β1), bone morphogenic protein 2 (BMP-2), dexamethasone (DEX), proline, ascorbic acid (ASA), pyruvic acid, insulin+transferrin+selenious acid pre-mixed solution (ITS), bovin serum albumin (BSA), basic fibroblast growth factor (bFGF), intermittent hydraulic pressure (IHP), bone morphogenic protein 7 (BMP-7), and insulin-like growth factor (IGF). The L60 (212) orthogonal experiment was designed using the SPSS 18.0 with 2 level conditions and the cells were induced to differentiate on the small intestinal submucosa (SIS)-3D scaffold. The CD151+/CD44+ cells were detected with the flow cytometry and then the differentiation rate was recorded. The immumohistochemical staining, cellular morphology, toluidine blue staining, and semi-quantitative RT-PCR examination for the gene expressions of sex determining region Y (SRY) -box 9 gene (Sox9), aggrecan gene (AGN), collagen type I gene (Col I), collagen type Ⅱ gene (Col Ⅱ), collagen type IX gene (Col IX) were used for result confirmation. The differentiation rate was calculated as the product of CD151/CD44+ cells and cells with Col I high expression. The grow curve was detected with the DNA abundance using the PicoGreen Assay. The visual observation and the variances analysis among the variable were used to evaluate the result of the orthogonal experiment, 1 level interaction was considered. The q-test and the least significant difference (LDS) were used for the variance analysis with a type Ⅲ calibration model. The test criteria (α) was 0.05.
ResultsThe cells were certified as SMSCs, the double-time of the cells was 28 hours. During the differentiation into the fibrocartilage, the volume of the SIS-3D scaffold enlarged double every 5 days. The scaffolds were positively stained by toluidine blue at 14 days. The visual observation showed that high levels of TGF-β1 and BMP-7 were optimum for the differentiation, and BMP-7 showed the interaction with BMP-2. The conditions of DEX, ASA, ITS, transferrin, bFGF showed decreasing promotional function by degrees, and the model showed the perfect relevance. P value was 0.000 according to the variance analysis. The intercept analysis showed different independent variables brought about variant contribution; the TGF-β1, ASA, bFGF, IGF, and BMP-7 were more remarkable, which were similar to the visual observation.
ConclusionIn the process of the SMSCs differentiation into the fibrocartilage, the concentrations of TGF-β1, ASA, bFGF, and IGF reasonably can improve the conversion rate of the fibrocartilage cells. The accurate conditions of the regulatory factor should be explored further.
【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.
ObjectiveTo study the effect of transforming growth factor β3 (TGF-β3), bone morphogenetic protein 2 (BMP-2), and dexamethasone (DEX) on the chondrogenic differentiation of rabbit synovial mesenchymal stem cells (SMSCs).
MethodsSMSCs were isolated from the knee joints of 5 rabbits (weighing, 1.8-2.5 kg), and were identified by morphogenetic observation, flow cytometry detection for cell surface antigen, and adipogenic and osteogenic differentiations. The SMSCs were cultured in the PELLET system for chondrogenic differentiation. The cell pellets were divided into 8 groups: TGF-β3 was added in group A, BMP-2 in group B, DEX in group C, TGF-β3+BMP-2 in group C, TGF-β3+DEX in group E, BMP-2+DEX in group F, and TGF-β3+BMP-2+DEX in group G; group H served as control group. The diameter, weight, collagen type II (immuohistochemistry staining), proteoglycan (toluidine blue staining), and expression of cartilage related genes [real time quantitative PCR (RT-qPCR) technique] were compared to evaluate the effect of cytokines on the chondrogenic differentiation of SMSCs. Meanwhile, the DNA content of cell pellets was tested to assess the relationship between the increase weight of cell pellets and the cell proliferation.
ResultsSMSCs were isolated from the knee joints of rabbits successfully and the findings indicated that the rabbit synovium-derived cells had characteristics of mesenchymal stem cells. The diameter, weight, collagen type II, proteoglycan, and expression of cartilage related genes of pellets in groups A-F were significantly lower than those of group G (P<0.05). RT-qPCR detection results showed that the relative expressions of cartilage related genes (SOX-9, Aggrecan, collagen type II, collagen type X, and BMP receptor II) in group G were significantly higher than those in the other groups (P<0.01). Meanwhile, with the increase of the volume of pellet, the DNA content reduced about 70% at 7 days, about 80% at 14 days, and about 88% at 21 days.
ConclusionThe combination of TGF-β3, BMP-2, and DEX can make the capacity of chondrogenesis of SMSCs maximized. The increase of the pellet volume is caused by the extracellular matrix rather than by cell proliferation.
Objective To explore the technique of arthroscopic treatment of synovial chondromatosis of the hip and to evaluate its effectiveness. Methods Between July 2009 and June 2011, 15 patients with synovial chondromatosis of the hip underwent arthroscopic synovectomy and removal of loose bodier. Of 15 patients, 11 were male and 4 were female, aged from 21 to 45 years with an average of 33.1 years. The location was the left side in 6 cases and the right side in 9 cases. The disease duration was 12-43 months (mean, 23 months) Pain and functional motion limitation were the main clinical symptoms. The visual analogue scale (VAS) score was 5.8 ± 1.1; the range of motion (ROM) of the hip was (149.8 ± 27.5)°; the Harris hip score was 54.5 ± 13.3. Results All incisions healed by first intention. All the patients were followed up 6 months to 2 years (mean, 17.4 months). At last follow-up, the VAS score was 2.0 ± 1.2; the ROM of the hip was (258.3 ± 35.4)°; the Harris hip score was 93.0 ± 18.7; and the above indexes were significantly improved when compared with preoperative values (P lt; 0.05). No recurrence was found on postoperative MRI. Conclusion Arthroscopic treatment of synovial chondromatosis of the hip has the advantages of minimal invasion, quick recovery, and best recovery of hip function and ROM.
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.
