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.
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.
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.
ObjectivesTo investigate the ultrasound findings of the synovial hemangioma of knee (SHK) and to evaluate its value in clinical diagnosis.MethodsThe ultrasonographic manifestations and clinical data of 10 patients with SHK confirmed by surgery and pathology were retrospectively analyzed and compared with MRI findings, surgery and pathological results.ResultsSeven cases of SHK (6 cases of diffuse type, 1 case of limited type) were assessed by ultrasound, including 1 case of vascular origin, 1 case of supraorbital sac origin, 1 case with pigmented villonodular synovitis, 1 case with thrombosis, 2 cases accompanied with bone erosion and osteophyte formation, and 3 cases with joint cavity effusion. Ultrasonic findings of SHK were as followed: 7 cases of SHK were manifestate as diffuse mass with unclear boundary, irregular shape and uneven echo mass; 5 cases had mixed-echo mass with reticular structures inside, an increased volume in erect position and positive CDFI compression test; 1 case had heterogeneous hypoechoic mass with a nodular appearance and the positive compression test; 1 case as poorly-demarcated, irregular shape, heterogeneous hyperechoic mass without obvious blood flow signals under the compression test. There were no characteristic ultrasonic findings from other 3 cases of SHK.ConclusionsDiffuse SHKs have characteristic ultrasonograms. SHKs with localized and significant synovial hyperplasia have no specific ultrasonic manifestation and are easily misdiagnosed. Ultrasound is convenient, noninvasive and inexpensive. It can accurately evaluate the involvement of knee joint capsule and surrounding soft tissues. It can be used as the first line diagnostic modality for routine scanning of SHKs.
Objective To evaluate the effectiveness of arthroscopy for synovial chondromatosis of hip joint. Methods Between April 2012 and September 2015, 32 patients with synovial chondromatosis of hip joint were treated by arthroscopy. There were 19 males and 13 females, with an average age of 42.1 years (range, 22-64 years). The synovial chondromatosis located at right hip in 15 cases and left hip in 17 cases. The main clinical symptoms were pain and swelling of hip joint. Of all patients, 6 cases were hip hinge, 2 cases were lower limb weakness, and 1 case was snapping hip. The " 4” sign was positive in 9 cases, Thomas’ sign positive in 4 cases, and rolling test positive in 2 cases. Results All incisions healed by first intention, and no complication occurred. All patients were followed up 16-48 months (mean, 33.8 months). The visual analogue scale (VAS) was 1.4±0.8 at last follow-up, which was significantly lower than that before operation (4.8±1.2) (t=6.382, P=0.013). The hip Harris score was 92.6±6.7 at last follow-up, which was significantly higher than that before operation (63.2±8.3) (t=9.761, P=0.006). At last follow-up, the " 4” sign and Thomas’ sign were positive in 3 cases and 1 case, respectively. The others had no positive sign. X-ray film showed no recrudescence in all cases. Conclusion Treating synovial chondromatosis of hip joint under arthroscopy has advantages of less trauma, complete debridement, quick postoperative recovery, and the satisfactory short-term effectiveness.
ObjectiveTo study the expressions of Renin, angiotensin converting enzyme (ACE), angiotensin receptor 1 (AT1R), and AT2R in synovial tissue of osteoarthritis (OA) at different stages.MethodsThe patients who were treated with upper knee amputation because of trauma or total knee arthroplasty for OA between January 2018 and December 2018 were enrolled. Among them, 32 patients who met the selection criteria were included in the study. According to the Kellgren-Lawrence (K-L) X-ray classification, they were allocated to normal synovial group (group A, n=9), moderate OA synovial group (group B, n=11, K-L level 3), and advanced OA synovial group (group C, n=12, K-L level 4). The relative expressions of Renin, ACE, AT1R, and AT2R mRNAs and proteins were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot.ResultsThe relative expressions of Renin, ACE, and AT1R mRNAs and proteins were significantly higher in group B and group C than in group A (P<0.05). The relative expressions of ACE and AT1R mRNAs and proteins and Renin protein were significantly higher in group C than in group B (P<0.05). However, the relative expressions of AT2R mRNA and protein were lower in group B and group C than in group A (P<0.05), and in group C than in group B (P<0.05).ConclusionThe expressions of Renin, ACE, and AT1R in synovial tissue of osteoarthritis significantly increase as the K-L level increased, and the expression of AT2R decreases. Renin, ACE, AT1R, and AT2R have a certain degree of correlation with the development of OA.
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.