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ABSTRACT: Osteosarcoma is the most common human primary malignant bone tumor in children and young adults. Sensitive and non-invasive biomarkers that can facilitate disease detection at early stage are highly desirable to improve survival rate and help to determine optimized treatment for osteosarcoma. The small non-coding RNAs, microRNAs (miRNAs), have recently been identified as critical regulators for various diseases including cancer and may represent a novel class of cancer biomarkers. In this study, we aimed to detect the potential of circulating miRNAs as biomarkers for osteosarcoma. Levels of six candidate miRNAs (miR-21, miR-199a-3p, miR-143, miR-34, miR-140, and miR-132) that were previously demonstrated to be regulated in osteosarcoma were examined in plasma of 40 osteosarcoma patients and 40 matched healthy controls by quantitative reverse-transcription polymerase chain reaction assays. The results showed that circulating levels of miR-21 were significantly higher in osteosarcoma patients than controls, while miR-199a-3p and miR-143 were decreased in osteosarcoma patients. We replicated the findings in an independent study of 40 osteosarcoma patients and 40 matched controls and confirmed the results. Receiver operating characteristics curve analysis of the combined populations demonstrated that the three-miRNA signature could discriminate cases from controls with an area under the curve of 0.953 (95 % CI 0.924-0.984). In addition, circulating miR-21 and miR-143 were correlated with both metastasis status and histological subtype of the patients, while miR-199a-3p only correlated with histological subtype. Our data suggest that altered levels of circulating miRNAs might have great potential to serve as novel, non-invasive biomarkers for osteosarcoma.
Medical Oncology 03/2013; 30(1):340. · 2.14 Impact Factor
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ABSTRACT: OBJECTIVES: Osteonecrosis is also known as avascular necrosis, and two types of cell death are included in the pathogenesis of osteonecrosis: necrosis and apoptosis. Apoptosis in the osteonecrosis of femoral head is thought to be the key determinant of glucocorticoid-induced cortical bone loss. The present study was implemented to evaluate the anti-apoptotic effect of Granulocyte colony-stimulating factor and stem cell factor (G-CSF/SCF) in rabbits with steroid-induced osteonecrosis. METHODS: In the experiment, osteonecrosis was induced by low-dose lipopolysaccharide and subsequent pulsed high-dose methylprednisolone. Rabbits in preventive medicine group were treated with100μg/kg/d G-CSF and 25μg/kg/d SCF. Then hematological and histomorphometric methods were used to investigate the treatment effects of osteonecrosis. Apoptosis was assessed via quantitative TUNEL staining and activated caspase-3 immunostaining and immunoblotting. RESULTS: The results showed that G-CSF/SCF treatment could increase the secretion of serum osteocalcin, but inhibit the expression of serum tartrate-resistant acid phosphatase (TRAP5b). The incidence of osteonecrosis was significantly decreased in Preventive group when compared with Steroid group (42.1% vs. 88.2%). Histomorphometric analysis showed that G-CSF/SCF pre-disposal treatment was able to increase trabecular mineral appositional rate (MAR) and bone formation rate (BFR). Quantitative TUNEL and activated caspase-3 levels showed lower apoptosis in the Preventive group. CONCLUSIONS: In conclusion, G-CSF/SCF treatment could inhibit caspase-3-dependent apoptosis in osteocytes to exert beneficial effects in preventing steroid-induced ON in rabbit models.
Experimental and Molecular Pathology 06/2012; · 2.42 Impact Factor
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ABSTRACT: By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein endothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of transplanted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdM(NOR), CdM-CdM(HYP) and HUVEC-CdM(HYP) were prepared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apoptosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cytometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. Compared with MSC-CdM(NOR) and HUVEC-CdM(HYP) groups, the survival rate, migration and angiogenesis of HUVECs in MSC-CdM(HYP) group were significantly increased while the apoptosis rate was declined (P<0.05). Moreover, the expression of SDF-1α, VEGF and IL-6 in MSC-CdM(HYP) group was up-regulated. Under hypoxia, the apoptosis of HUVECs was inhibited while survival, migration and angiogenesis were improved by co-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.
Journal of Huazhong University of Science and Technology 04/2012; 32(2):173-80. · 0.38 Impact Factor
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ABSTRACT: The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells. The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α, and the expression of β-catenin protein was detected by Western blotting. The cultured rabbit nucleus pulposus cells were divided into 4 groups. In group A, the cells were cultured with normal medium and served as control group. In group B, the cells were cultured with TNF-α and acted as degeneration group. In group C, the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group. In group D, the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP, serving as intervention group. The expression of type II collagen, proteoglycan, β-catenin, and MMP-13 in each group was detected by immunocytochemistry and RT-PCR. The result showed that TNF-α increased the expression of β-catenin and MMP-13, and significantly inhibited the synthesis of type II collagen and proteoglycan, which resulted in the degeneration of nucleus pulposus cells. This effect could be obviously reversed by DKK1. We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway, and increase the expression of MMP-13, thereby resulting in disc degeneration. Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue. The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.
Journal of Huazhong University of Science and Technology 10/2011; 31(5):657-62. · 0.38 Impact Factor
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ABSTRACT: Previous studies have shown that novel scaffold-free self-assembled constructs can be an ideal alternative for cartilage tissue engineered based on scaffolds, which has many limitations. However, many questions remain, including the choice of seeding cells and the role of growth differentiation factor 5 (GDF-5) in constructing self-assembled engineered cartilages. Moreover, whether the optimum construct is effective in human chondral defect repair is still unknown. In this study, we generated self-assembled constructs of human mesenchymal stem cells (hMSCs) using four different approaches: direct self-assembly of hMSCs with or without GDF-5, and predifferentiated hMSCs self-assembly with or without GDF-5. Histological, immunohistochemical, and biochemistry analyses indicated that the constructs generated from predifferentiated hMSCs induced by GDF-5 (Group D2) exhibited up-regulated glycosaminoglycan (GAG) and type II collagen expression and contained higher amounts of GAG and total collagen than any other group. After 3-weeks of in vitro culturing of the constructs in a chondral defects explant culture system, the contructs from Group D2 were stably adhered to the surface of the cartilage matrix. Immunohistochemically, the repair tissue was positive for type II collagen, toluidine blue, and safranin O. These data demonstrated that the generation of self-assembled tissue-engineered cartilage from chondrogenically differentiated hMSCs induced by GDF-5 is a promising therapeutic strategy for cartilage repair.
Tissue Engineering Part C Methods 08/2011; 17(12):1189-99. · 4.64 Impact Factor
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ABSTRACT: To study the effects of zoledronic acid (ZA) on the vasculogenic mimicry of osteosarcoma cells in vitro.
A Three-dimensional culture of LM8 osteosarcoma cells on a type I collagen matrix was used to investigate whether osteosarcoma cells can develop vasculogenic mimicry, and to determine the effects of ZA on this process. In addition, the cellular ultrastructural changes were observed using scanning electron microscopy and laser confocal microscopy. The effects of ZA on the translocation of RhoA protein from the cytosol to the membrane in LM8 cells were measured via immunoblotting.
ZA inhibited the development of vasculogenic mimicry by the LM8 osteosarcoma cells, decreased microvilli formation on the cell surface, and disrupted the F-actin cytoskeleton. ZA prevented translocation of RhoA protein from the cytosol to the membrane in LM8 cells.
ZA can impair RhoA membrane localization in LM8 cells, causing obvious changes in the ultrastructure of osteosarcoma cells and induce cell apoptosis, which may be one of the underlying mechanisms by which the agent inhibits the development of vasculogenic mimicry by the LM8 cells.
BMC Musculoskeletal Disorders 06/2011; 12:146. · 1.58 Impact Factor
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ABSTRACT: To evaluate the therapeutic results of hemivertebra excision through combined anterior and posterior approach followed by short-segment spinal fusion with transpedicular instrumentation in children. Clinical and radiological evaluations were conducted before and after operation and at the follow-up examination. Cobb angles of segmental curve, total main curve, compensatory curves (cranial and cauda) and kyphosis were recorded. All the cases showed satisfactory results, the mean Cobb angle of segment curve was 45.3° before surgery and 14.9° after surgery with 67.1% correction, and 13.5° at the final follow-up assessment with 70.2% correction. Total main curves improved from 47.6 to 18.2° with 61.8% correction, and 16.0° at the final follow-up. Cranial and caudal compensatory curves were improved gradually during follow-up. Kyphosis of the spine also improved obviously. If the operation was performed before the maturity of bone, the secondary spinal deformities could be effectively avoided.
Journal of pediatric orthopaedics. Part B / European Paediatric Orthopaedic Society, Pediatric Orthopaedic Society of North America 11/2010; 19(6):545-50. · 0.66 Impact Factor
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ABSTRACT: The aim of this study was to evaluate the effect of a porous tantalum rod implant for the treatment of early femoral head necrosis. From April 2007 to June 2009, a total of 35 femoral head necrosis patients (with 49 hips) were treated with core decompression in combination with the insertion of a porous tantalum rod. The mean age was 38.2 years (22-50 years) and the mean follow-up period was 15.2 months (12-36 months). The surgical time and blood loss were recorded. The Harris hip scores and radiological results were adopted for evaluation. The mean surgical time was 35 min, and the mean blood loss was 50 ml. The mean Harris score improved from 48.3 ± 3.2 preoperative to 83.7 ± 4.1 at the last follow-up (p < 0.05). Eight affected hips exhibited progressive pain including three hips that progressed to femoral collapse, and one revision followed by total hip arthroplasty (THA). For the patient who underwent revision and THA, the articular cartilage surface was seen to be damaged and fragmented. High-density metal particle residuals were observed on radiograph in the bone channel and femoral marrow cavity. We conclude that the selection criteria for porous tantalum implants should be early and intermediate stages of femoral head necrosis. Further study is warranted to reveal whether the metal particles released play a role in the progression of pain and failure.
Biomedical Materials 10/2010; 5(6):065003. · 2.16 Impact Factor
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ABSTRACT: Prospective cohort study.
To comprehensively compare the intermediate and long-term results of posterior correction and fusion with segmental pedicle screw instrumentation versus those with hook constructs in idiopathic adolescent thoracic scoliosis.
Posterior correction and fusion represent the current standard surgical treatment in progressive idiopathic thoracic scoliosis. The 3-column fixation of pedicle screws has been shown to be superior to all other posterior spinal fixation devices.
A total of 168 patients with idiopathic thoracic scoliosis at a single institution who underwent a posterior spinal fusion with segmental pedicle screw (88) or a combination of hooks and pedicle screws (80) instrumentation. Patient's evaluation consisted of clinical and radiographic analysis preoperatively, postoperatively, and at final follow-up.
All patients were prospectively evaluated with an average follow-up of 5 years (range 5 to 11 y). The average number of segments in the fusion was 9.1 (range 6 to 15) in the hook group and 8.5 in the screw group (range 5 to 12). At the final follow-up, the amount of loss of correction in thoracic curves averaged 8.4 in the hook group and 5.3 in the screw group. The difference between the mean postoperative Cobb angle and the final Cobb angle of the major curves with a preoperative value was statistically significant in the 2 groups (P<0.01). The frontal and sagittal plane correction can be satisfactorily obtained by the screw group versus the pedicle screw group. There were no cases of pseudarthrosis, deep wound infections, or any neurologic complications.
Satisfactory correction and maintenance of scoliotic curves could be obtained by pedicle screw instrumentation compared with hook constructs. Thoracic and thoracolumbar pedicle screw instrumentation is a safe and reliable method for obtaining rigid segmental spinal fixation over the conventional hook and rod.
Journal of spinal disorders & techniques 10/2010; 23(7):467-73. · 1.21 Impact Factor
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ABSTRACT: To evaluate the short-term clinical outcomes of metal-on-metal total hip resurfacing arthroplasty in treating osteonecrosis of the femoral head (ONFH) in young and middle-aged patients and to compare with patients of hip osteoarthritis at the same period.
From July 2006 to October 2008, 33 patients (45 hips) with ONFH (ONFH group) and 39 patients (45 hips) with osteoarthritis (osteoarthritis group) were treated with metal-on-metal total hip resurfacing arthroplasty. In ONFH group, there were 18 males (27 hips) and 15 females (18 hips) with an average age of 42.7 years (range 19-58 years), including 22 left hips and 23 right hips. The causes were trauma (4 cases), glucocorticoid (25 cases), drugs (2 cases), alcohol (1 case), and psoriasis (1 case). According to Steinberg classification, there were 10 hips at stage III, 18 hips at stage IV a, 13 hips at stage IV b, and 4 hips at stage IV c. The Harris score was 52.0 +/- 4.6. The disease course was 1-12 years. In osteoarthritis group, there were 26 males (30 hips) and 13 females (15 hips) with an average age of 47.1 years (range 42-65 years), including 17 left hips and 28 right hips. The causes were degenerative arthritis (23 cases), trauma (11 cases), and ankylosing spondylitis (5 cases). The Harris score was 57.0 +/- 3.8. The disease course was 3-17 years. There were no significant differences in general data between two groups (P > 0.05).
All incisions achieved healing by first intention without complications of infection and thrombosis of deep vein of lower extremities. The patients were followed up for 26 months in ONFH group and 28 months in osteoarthritis group. Femoral neck fracture occurred in 1 case of osteoarthritis group after 4 months, who received total hip arthroplasty; no complication of prosthesis loosening, dislocation, incision infection, osteonecrosis, and bone absorption occurred in other patients. At last follow-up, the Harris scores were 93.0 +/- 5.5 in ONFH group and 94.0 +/- 2.4 in osteoarthritis group, showing no significant difference between two groups (P > 0.05); but there were significant differences between pre- and post-operation (P < 0.01).
The clinical short-term outcomes of metal-on-metal total hip resurfacing arthroplasty to treat ONFH are satisfactory. It can achieve similar outcomes to that of osteoarthritis group. More cases and long-term follow-up are needed to investigate long-term clinical outcomes.
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery 03/2010; 24(3):262-5.
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ABSTRACT: Seventy-six patients with femoral head necrosis were allocated to a program of either core decompression (control group) or core decompression and implantation of a biomaterial-loaded allograft threaded cage (treatment group). All patients were followed up prospectively clinically and radiographically. In the control group, no significant improvement in Harris hip score was found, and 13 of the 22 hips had deteriorated to stage III. In the treatment group, the mean Harris hip score was improved from 62.8 to 81.6; the clinical success rate at 36 months postoperatively was 91%. Collapse was seen in 1 hip, and another 3 hips exhibited progressive collapse. The procedure is attractive as a minimally invasive and salvage procedure, which shows encouraging success rates and early clinical results in patients with Steinberg stage I-II osteonecrosis.
The Journal of arthroplasty 10/2009; 25(8):1223-30. · 1.79 Impact Factor
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ABSTRACT: To investigate clinical therapeutic effect on early stage femoral head necrosis managed with allogeneic cortical bone cage support combining with autologous cancellous bone grafting through core decompression tunnel, and to discuss its effect on preventing femoral head collapse and influence factors.
From January 2002 to December 2005, 40 patients (42 hips) with femoral head necrosis underwent core decompression and an allogeneic threaded cortical bone supporting cage which was loaded with autologous cancellous bone inside. There were 26 males and 14 females, aging 27-45 years (mean 35.6 years). The disease course was 6-28 months (mean 18.3 months). All the cases underwent X-ray, CT and MRI examination to confirm the diagnosis and necrosis area. Twelve hips were at Ficat stage I, 29 hip at stage II, and 1 hips at stage III. Harris hip score system was used to evaluate the hip function pre- and post-operatively. X-ray films were taken regularly after operation.
All the wound healed by first intention without any complications such as infection, fracture, nerve and vascular injury, and deep vein thrombosis. Thirty-six patients (38 hips) were followed up for 24-58 months with an average of 38 months. All the patients had different degrees of improvement in clinical symptoms. According to Harris hip score system, the Harris score was 63.1 +/- 6.4 before operation and 82.3 +/- 16.5 at the last follow-up, showing significant difference (P < 0.001). The results were excellent in 24 hips, good in 11 hips, fair in 2 hips and poor in 1 hips. The X-ray films showed femoral head repairing and no advancement of osteonecrosis and collapsing in 23 patients (24 hips) 24 months after operation.
Allogeneic cortical bone cage support combining with autologous cancellous bone grafting is suitable for managing early stage femoral head necrosis and its short- and middle-term effect is satisfactory.
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery 05/2009; 23(5):527-9.
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ABSTRACT: The purpose of this study was to evaluate the efficacy of core decompression with a biomaterial-loaded allograft threaded cage (ATC) for the treatment of femoral head osteonecrosis in an established goat model. First, bilateral early-stage osteonecrosis was induced. After core decompression, the remaining goats were randomly divided into three groups: Group A, the goats were left without any treatment; Group B, the goats were treated with implanting a composite of autologous bone and decalcified bone matrix (DBM); Group C, the goats were treated using insertion of ATC loaded with DBM and autogenous bone graft. Then radiographic, histological and biomechanical analysis were taken in each group at 5, 10, and 20 weeks postoperation. In Group A, the classical signs of osteonecrosis of the femoral head were identified 10 weeks after the induction. Twenty weeks later, the density, surface and biomechanical stability of the femoral head were normal in Group C, while an irregular surface and an inhomogeneous microstructure or variation of density/hardness were identified in Group B. The specimens revealed a continuous trabaecular bone structure throughout the cage and extensive bone ingrowth and remodeling in Group C, while fibrous tissue was evident in Group B. Core decompression with a biomaterial loaded ATC almost uniformly delays or arrests the progression of the disease before articular collapse, and it could help to get the balance between bone resorption and new bone formation, strengthen structural mechanics of the femoral head, provide structure support of articular cartilage.
Biotechnology and Bioengineering 07/2008; 100(3):560-6. · 3.95 Impact Factor
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ABSTRACT: Our clinical study design was prospective, concurrently enrolled and single-center trial of the new functional intervertebral cervical disc prosthesis (Medtronic Sofamor Danek, Memphis, TN) in the treatment of patients with single-level and multiple-level degenerative disc disease of the cervical spine.
The study was designed to investigate the surgical technology skills and clinical effects of Bryan cervical disc prosthesis in Chinese, and to observe the stability and range of movement in the early and immediate postoperative period.
Recently, motion preservation has come to the forefront of emerging technologies in spine surgery. This is the important background information of the emergence of cervical arthroplasty as an alternative to arthrodesis that offers the promise of restoring normal spinal movement and reduces a kinematic strain on adjacent segments.
Nineteen patients (23 discs) had spinal arthroplasty with placement of the Bryan cervical artificial disc prosthesis in our hospital. Clinical and radiologic follow-up was performed. The radiographic parameters evaluated included the treated segments and the overall curvature of the cervical spine.
All the patients were observed up from 8 months to 42 months (average 24 months). According to Odom's scale, all of 19 patients (23 levels) had excellent to good outcome. The range of movement recovered to the preoperative value during the follow up. The treated segment ultimately showed preservation of movement when compared with preoperative levels. No prosthesis subsidence or excursion was identified.
Arthroplasty using the Bryan disc seemed to be safe and provided encouraging clinical and radiologic outcome in our study. Although early and intermediate results are promising, this is also a relatively new approach, long-term follow up studies are required to prove its efficacy and its ability to prevent adjacent segment disease.
Spine 06/2008; 33(12):E371-7. · 2.08 Impact Factor
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ABSTRACT: To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Journal of Huazhong University of Science and Technology 09/2007; 27(4):429-32. · 0.38 Impact Factor
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ABSTRACT: This study examined effect of a new intervertebral cervical disc prosthesis in relieving the neurological symptoms and signs, improving the patients' ability to perform daily activities, reducing pain, and maintaining the stability and segmental motion. From December 2003 to October 2004, 12 patients, who had received 14 replacements of cervical artificial discs, were followed-up for 2 to 8 months (with a mean of 5.2 months). Of them 5 had cervical spondylotic myelopathy and 7 had cervical disc herniation. The patients included 7 males and 5 females, with their age ranging from 35 to 62 y and a mean of 50.3 y. Single-level replacements were performed in 10 cases and 2 cases received two-level replacement. Operation time of the single-level surgery averaged 130+/-50 min and the time of two-level surgery was 165+/-53 min on average (from skin incision to skin suturing). Neurological or vascular complications during or after surgery was not observed. Japanese Orthopedic Association scores (JOA scores) increased from 8.6 to 15.8 on average. There was no prothesis subsidence or excursion. Replaced segments were stable and the range of motion was partially restored, being 4.68 degrees (3.6 degrees -6.1 degrees ) in flexion and extension position and 3.51 degrees (2.5 degrees -4.6 degrees ), 3.42 degrees (2.6 degrees -4.3 degrees ) in left and right bending position. No obvious loss of physiological curvature was noted. CT or MRI follow-up showed that excursion was less than 1.5 mm) in 2 of 14 levels and between 1.5 mm and 3 mm) in 1 of 14 levels. No ossification in the replaced levels was observed. It is concluded that satisfactory short-term results were achieved in the 12 cases of artificial disc replacements. Different from anterior cervical discectomy and fusion, the replacement could achieve quick functional recovery and did not lead to the movement limitation of cervical vertebrae. At least a 5-years follow-up was needed to assess the long-term effect of the prosthesis on its neighboring segments.
Journal of Huazhong University of Science and Technology 05/2007; 27(2):176-8. · 0.38 Impact Factor
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ABSTRACT: The biocompatibility and osteogenic activity of allogenic decalcified bone matrix (DBM) used as a carrier for bone tissue engineering were studied. Following the method described by Urist, allogenic DBM was made. In vitro, DBM and bone marrow stromal cell (BMSC) from rabbits were co-cultured for 3-7 days and subjected to HE staining, and a series of histomorphological observations were performed under phase-contrast microscopy and scanning electron microscopy (SEM). In vivo the mixture of DBM/BMSC co-cultured for 3 days was planted into one side of muscules sacrospinalis of rabbits, and the DBM without BMSC was planted into other side as control. Specimens were collected at postoperative week 1, 2 and 4, and subjected to HE staining, and observed under SEM. The results showed during culture in vitro, the BMSCs adherent to the wall of DBM grew, proliferated and had secretive activity. The in vivo experiment revealed that BMSCs and undifferentiated mesenchymal cells in the perivascular region invaded gradually and proliferated together in DBM/BMSC group, and colony-forming units of chondrocytes were found. Osteoblasts, trabecular bone and medullary cavity appeared. The inflammatory reaction around muscles almost disappeared at the second weeks. In pure DBM group, the similar changes appeared from the surface of the DBM to center, and the volume of total regenerate bones was less than the DBM/BMSC group at the same time. The results indicated that the mixture of DBM and BMSC had good biocompatibility and ectopic induced osteogenic activity.
Journal of Huazhong University of Science and Technology 02/2004; 24(2):147-50. · 0.38 Impact Factor
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ABSTRACT: To explore a new method for the therapy of avascular necrosis of the femoral head.
The recombinant plasmid pCD-hVEGF165 was mixed with collagen and was implanted in the necrotic femoral head. The expression of vascular endothelial growth factor (VEGF) was examined by RNA dot hybridization and immunohistochemical techniques. Repair of the femoral head was observed by histological and histomorphometric analysis.
The expression of VEGF was detected in the femoral head transfected with the VEGF gene. The femoral head transfected with the VEGF gene showed a significant increase in angiogenesis 2 and 4 weeks after gene transfection and a significant increase in bone formation 6 and 8 weeks after gene transfection on histomorphometric analysis (P < 0.01).
Transfection of the VEGF gene enhances bone tissue angiogenesis. Repair of osteonecrosis could be accelerated accordingly, thus providing a potential method for therapy of osteonecrosis.
Chinese medical journal 10/2003; 116(10):1544-8. · 0.86 Impact Factor
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ABSTRACT: To explore a new method for the therapy of the avascular necrosis of the femoral head, the recombinant plasmid pCD-hVEGF165 was mixed with collagen and was implanted in the necrotic femoral head. The expression of vascular endothelial growth factor (VEGF) was detected by RNA dot hybridization and immunohistochemical method. The repair of the femoral head was observed by histological method. The results showed that the expression of VEGF was detectable in the femoral head treated with VEGF gene. Angiogenesis in these femoral heads was more abundant than the control. Bone repairing was augmented in the femoral head treated with VEGF gene. The results suggest that angiogenesis in bone tissue could be augmented by gene transfection of VEGF and bone repairing would be accelerated accordingly.
Journal of Huazhong University of Science and Technology 02/2003; 23(3):297-9, 316. · 0.38 Impact Factor
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ABSTRACT: ObjectiveTo investigate the effect of the gap junction blocker 1-heptanol on the in vitro chondrogenic differentiation of mouse bone marrow mesenchymal stem cells(MSCs) following induction by GDF-5.MethodsMSCs were isolated from mouse bone marrow and cultured in vitro. After 3 passages cells were induced to undergo chondrogenic differentiation with recombinant human GDF-5(100 ng/ml), with or without 1-heptanol(2.5μ mol/L). The effect of 1-heptanol on MSCs proliferation was investigated using the MTT assay. Type, collagen mRNA and protein were examined by RT-PCR and immunocytochemistry respectively, and the sulfate glycosaminoglycan was assessed by Alcian blue dye staining. Connexin43(Cx43) protein was examined by western blotting.ResultsGDF-5 induced proliferation and chondrogenic differentiation of MSCs. While 1-heptanol treatment had no effect on this proliferation, it inhibited the expression of both type, collagen mRNA and protein. The Alcian blue staining revealed that 1-heptanol also inhibited the deposition of the typical cartilage extracellular matrix promoted by recombinant GDF-5. Western blotting demonstrated that 1-heptanol had no effect on the expression of Cx43.ConclusionThese results suggest that mouse bone marrow MSCs can be differentiated into a chondrogenic phenotype by GDF-5 administration in vitro. While the gap junction blocker, 1-heptanol, did not reduce gap junction Cx43, these intercellular communication pathways clearly played an important functional role in GDF-5-induced cartilage differentiation.
Journal of Nanjing Medical University.
