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BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned. OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks. RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.
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Objective To examine the gene expression profile of SRY-type high-mobility-group box-9(SOX9) during entochondrostosis of mice and investigate the effects of transfection of the pDC316-SOX-9 on mice mesenchymal stem cells(MSCs) in vitro. Methods cDNA microarray technique with 34 000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of SOX9 was performed with GCOS1.2 software. The recombined expression vector pDC316-SOX-9 was constructed and transfected into mice MSCs by lipofectamine. The phenotype changes of cells were observed with cell energometry, HE stain, immunohistochemical method, RT-PCR and ELISA.Results The gene expression of SOX9 during the critical phase of chondrogenesis in mice embryo limbs at E12 was increased evidently. SOX9 might promot chondrogenesis. As compared with vector and blank group,the chondrocytes of the SOX9 transfected group had the tendency of enhanced differentiation. Conclusion SOX9 may promote chondrogenesis. The transfection of SOX9 gene into mice MSCs can promot MSCs differentiate into chondrocyte, which may provide some experimental data for cartilage histoengineering.
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To investigate the efficacy of a combination therapy on gluteal muscle contracture, 286 definitely diagnosed patients were subjected to surgical treatment, and then functional exercises and physical therapy. The patients with severe symptoms were asked to have a set of specially-designed functional exercises. All the patients were followed up for 3 to 24 months by hospital visit, correspondence or telephone interview. The effective rate was 100%, and the curative rate was up to 94.6%. Few patients developed complications and relapse was rare. It is concluded that the combination therapy, including surgical removal of diseased tissues, functional exercises and physical therapy, is an effective approach for the treatment of severe juvenile gluteal muscle contracture.
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Buttocks , Exercise Therapy , Follow-Up Studies , Muscle Contraction , Muscle, Skeletal/pathology , Patient Compliance , Physical Therapy Modalities , Time Factors , Treatment OutcomeABSTRACT
A retrospective analysis was performed among 8 patients with unilateral intraarticular calcaneal fracture, who were selected from the Department of Orthopaedics in Yangpu District Central Hospital of Shanghai. They were treated with the injectable bone graft MIIG[R] X3 combined with percutaneous reduction by leverage and axial fixation from June 2005 to November 2006. After treatment, the function of affected feet was evaluated according to the ankle-hindfoot score of the American Orthopaedic Foot & Ankle Society (totally 100 points). All the patients were rechecked with X-ray films periodically to observe the reaction between the materials and the host, the healing of fracture, the pain of limbs and joint motion. Eight patients were all followed up, the complication rate reached 12.5% (1/8), the improvement of Bohler angle excelled 18° to 25°, and that of Gissane angle exceeded 30° to 35°. Among the included patients, 6 cases achieved excellent effect and 2 cases good. All developed the bone union. None of all had complications, such as necrosis of skin, hematoma and deep infection.
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0.05).CONCLUSION:The excellent mechanical properties of composite ligament can meet the mechanical requirements of appropriate ligament tissue engineering scaffolds.
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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.
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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° (3.6°-6.1°) in flexion and extension position and 3.51° (2.5°-4.6°) 3.42° (2.6°-4.3°) 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.
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[Objective]To evaluate surgical outcomes of hemivertebra resection only via posterior approaches followed by short-segment transpedicular instrumentation for correction of the congenital scoliosos. [Method]A group of patients with a single hemivertebra between the ages 5-16 years who underwent operative treatment were evaluated.Hemivertebra resection through posterior approach and short-segment transpedicular instrumentation were used for correction of the scoliosis and kyphosis deformities.Radiographic evaluation were conducted before and after operation and at follow-up visits.Cobb's angles of the segmental curve,total main curve,cranial and caudal compensatory curves and kyphosis were measured on the posteroanterior and lateral radiographs.[Result]The cases in this study showed satisfied results.The mean Cobb's angle of segment curve was 41.5? before surgery and 15.1? after surgery with a 63.6% correction,and 14.7? at the lastest follow-up assessment with a 64.6% correction;total main curves improved from 46.9?to 18.4?,with a 60.8% correction;17.5?at last follow-up,with a 62.7% correction;the correction ratio for kyphosis was from 15.4?kyphosis to normal physical profile;cranial and caudal compensartory curves were obviously improved.[Conclusion]Hemivertebra resection and short-segment transpedicular instrumentation could be performed only through posterior approach,which had exerted satisfactory correction on congenital scoliosis.If the operation is performed before the maturity of the bone,it will effectively prevent the formation of secondly spinal deformities.
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Lack of biocompatibility and bioactivity is a big problem for the synthetic materials that have been generated for neural tissue engineering. To get around the problem and generate better scaffold for neural tissue repair, we intended to generate nano-fibers by self-assembly of polypeptide IKVAV. Bioactive IKVAV Peptide-Amphiphile (IKVAV-PA) was first synthesized and purified, the property of which was analyzed and determined by high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Then, by addition of hydrogen chloride (HCl), self-assembly of IK-VAV-PA was induced in vitro and nano-fibers formed as shown by transmission electron microscopy (TEM). The effect of IKVAV nanofibers on adherence of PC12 cells was assayed in cell culture and the results showed that the rates of adherence of PC12 increased significantly when the density of IKVAV was within a certain range (0.58 microg/cm2 to 15.6 microg/cm2). However, its effect on the rates of adherence did not significantly alter with time, whether after 1 hour or 3 hours of culture. In general, we showed that IKVAV-PA can successfully self-assemble to form nanofiber, and promote rapid and stable adherence of PC12 cells, and the effect of the self-assembled IKVAV to promote PC12 cells adherence is dosage-dependent within a certain range of densities.
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The necessity and superiority of the surgical operation on children with floating knee injury and the fracture union and complications were investigated. Twenty-eight children with floating knee injury were subjected to open reduction and internal fixation or external fixator. The patients were followed up for 18 months to 7 years. The curative effectiveness was scored by Karlstrom criteria. The results showed that no nonunion or deformity was found. The affected limb was 1.2 cm to 1.5 cm longer in 2 cases, 0.8 to 1.2 cm shorter in 3 cases than the contralateral. No severe dysfunction of knee joint occurred. The excellent-good rate was 92.8% and the curative rate 71.4% respectively. So for children whose age is older than 5 years, it's a good way to treat the fractures of femur and tibia with open reduction and internal fixation or external fixator. The method can be advantageous for the nursing care, early function recovery, shortening of the hospital stay and avoidance of severe complications.
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Femoral Fractures/complications , Femoral Fractures/surgery , Fracture Fixation , Knee Injuries/classification , Knee Injuries/etiology , Knee Injuries/surgery , Tibial Fractures/complications , Tibial Fractures/surgery , Treatment OutcomeABSTRACT
The necessity and superiority of the surgical operation on children with floating knee injury and the fracture union and complications were investigated. Twenty-eight children with floating knee injury were subjected to open reduction and internal fixation or external fixator. The patients were followed up for 18 months to 7 years. The curative effectiveness was scored by Karlstrom criteria. The results showed that no nonunion or deformity was found. The affected limb was 1.2 cm to 1.5 cm longer in 2 cases, 0.8 to 1.2 cm shorter in 3 cases than the contralateral. No severe dysfunction of knee joint occurred. The excellent-good rate was 92.8 % and the curative rate 71.4 % respectively. So for children whose age is older than 5 years, it's a good way to treat the fractures of femur and tibia with open reduction and internal fixation or external fixator. The method can be advantageous for the nursing care, early function recovery, shortening of the hospital stay and avoidance of severe complications.
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Lack of biocompatibility and bioactivity is a big problem for the synthetic materials that have been generated for neural tissue engineering. To get around the problem and generate better scaffold for neural tissue repair, we intended to generate nano-fibers by self-assembly of polypeptide IKVAV. Bioactive IKVAV Peptide-Amphiphile (IKVAV-PA) was first synthesized and purified, the property of which was analyzed and determined by high-performance liquid chromatography (HPLC)and mass spectrometry (MS). Then, by addition of hydrogen chloride (HCl), self-assembly of IKVAV-PA was induced in vitro and nano-fibers formed as shown by transmission electron microscopy (TEM). The effect of IKVAV nanofibers on adherence of PC12 cells was assayed in cell culture and the results showed that the rates of adherence of PC12 increased significantly when the density of IKVAV was within a certain range (0.58 μg/cm2 to 15.6 μg/cm2). However, its effect on the rates of adherence did not significantly alter with time, whether after 1 hour or 3 hours of culture. In general,we showed that IKVAV-PA can successfully self-assemble to form nanofiber, and promote rapid and stable adherence of PC12 cells, and the effect of the self-assembled IKVAV to promote PC12 cells adherence is dosage-dependent within a certain range of densities.
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Objective To investigate the efficacy of transcutaneous injection of MIIGX3 artificial bone and methylprednisolone for recurrent bone cysts.Methods From January 2004 to March 2006,a total of 13 children with recurrent bone cysts received transcutaneous injection of methylprednisolone and MIIGX3 in our hospital.X-ray was employed to detect the degradation of MIIGX3 and formation of new bones.Results The operation time ranged from 30 to 95 minutes(mean 42 minutes).The patients were followed up for 1 to 3 years.None of them had recurrence of bone cysts during this period.The artificial bones were completely degraded and replaced by new bones in 1.5 years after the injection.Conclusions Transcutaneous injection of methylprednisolone and MIIGX3 is effective for recurrent bone cysts.Patients have shorter operation time and hospital stay after this treatment because the procedure is microinvasive.
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BACKGROUND: The cells of biomaterial compound cytokine gene or compound transfected cytokine gene that are transplanted into the area of bone defect can promote bone repair.OBJECTIVE: To investigate the feasibility of gene therapy for bone defect after rat transforming growth factor (TGF)β1 gene is transfected into osteoblasts.DESIGN: A controlled and observational experiment.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Five newborn Sprague-Dawley rats of either gender were included.METHODS: The experiment was conducted in the laboratory of Orthopedic Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, from February to September 2000.Rat osteoblasts were transfected with pcDNA3-TGF-β1 plasmid by lipofectamine mediated gene transfer, and the plasmid pcDNA transfected cells were set as control group. The transient expression of TGF-β1 was detected by strept avidin-biotin-peroxidase complex (SABC) method and in situ hybridization detection 24 hours later. The cells transfected by G418 for 2 weeks were detected with SABC to investigate the stable expression of TGF-β1 gene.MAIN OUTCOME MEASURES: SABC method and in situ hybridization detection were applied to detect gene expression.of pcDNA3-TGF-β1 transfected osteoblasts and in situ hybridization detection: After the osteoblasts were transfected for 24 hours, cytoplast was full of brown granules and there were no brown granules in the cytoplast of blank carrier transfected cells, indicating that TGF-β1 mRNA was signifiG418: transfected cells still had high expression of TGF-β1 after 2-week G418 screening.CONCLUSION: Osteoblasts can express cytokine immediately with high effect using gene transfection technique. The stable and high expression is presented after TGF-β1 gene is transfected into osteoblasts at the moment of transfection and after 2-week screening, proving the feasibility of gene therapy for bone defect when cytokine gene is transfected into osteoblasts.
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To investigate therapeutic efficiency of Ad/CMV- hTGF-beta1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the 1.5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral disc degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 microl (10 x 10(6) pfu) of Ad/CMV- hTGF-beta1 gene, constituted the therapy group, 12 were injected with 20 microl (10 x 10(6) pfu)of Ad/CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group, 3 weeks after injection, examples were taken for investigation of HE staining, MRI, Western Blotting and immunohistochemical research TGF-beta1. Wide distribution of TGF-beta1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-beta1 content in intervertebral discs treated with TGF-beta1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral disc pathological degree improved. Ad/CMV- hTGF-beta1 gene transfection is a potential method to increase TGF-beta1 content and reverse intervertebral disc degeneration.
Subject(s)
Adenoviridae/genetics , Genetic Therapy , Gene Transfer Techniques , Genetic Vectors , Intervertebral Disc/pathology , Lumbar Vertebrae , Spinal Diseases/drug therapy , Transfection , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta1ABSTRACT
To investigate therapeutic efficiency of Ad/CMV- hTGF-beta1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the 1.5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral disc degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 microl (10 x 10(6) pfu) of Ad/CMV- hTGF-beta1 gene, constituted the therapy group, 12 were injected with 20 microl (10 x 10(6) pfu)of Ad/CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group, 3 weeks after injection, examples were taken for investigation of HE staining, MRI, Western Blotting and immunohistochemical research TGF-beta1. Wide distribution of TGF-beta1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-beta1 content in intervertebral discs treated with TGF-beta1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral disc pathological degree improved. Ad/CMV- hTGF-beta1 gene transfection is a potential method to increase TGF-beta1 content and reverse intervertebral disc degeneration.
Subject(s)
Animals , Rabbits , Adenoviridae , Genetics , Gene Transfer Techniques , Genetic Therapy , Genetic Vectors , Intervertebral Disc , Pathology , Lumbar Vertebrae , Spinal Diseases , Drug Therapy , Transfection , Transforming Growth Factor beta , Genetics , Transforming Growth Factor beta1ABSTRACT
Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P
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Objective To determine whether rabbit articular chondrocytes express growth factor genes delivered by recombined rat TGF beta 1,IGF 1 and what other influences on chondrocytes are,and to determine which gene is the best one for osteoarthritis therapy.Methods Monolayer cultures of rabbit articular chondrocytes were infected with recombinant plasmid pcDNA 3 and pAT 153 carrying genes encoding the following growth factors respectively:TGF beta 1 and IGF 1.The synthesis of TGF beta 1,IGF 1 and type Ⅱ collagen was measured by in situ hybridization,immunohistochemistry,immunofluoroscopy,flow cytometer and 3 H TdR radiolabeling.Results The expression of TGF beta1,IGF 1 and type Ⅱ collagen was high beyond control levels ( P
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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.
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Animals , Rabbits , Collagen , Therapeutic Uses , Femur Head , Femur Head Necrosis , Therapeutics , Gene Transfer Techniques , Genetic Therapy , Implants, Experimental , Neovascularization, Physiologic , Osteogenesis , Plasmids , Genetics , Random Allocation , Recombinant Proteins , Genetics , Vascular Endothelial Growth Factor A , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore a new method for the therapy of avascular necrosis of the femoral head.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>