Objective To observe the treatment effect of transplanted autogenous bone marrow stroma cells (BMS-C) absorbed in the type-Ⅰ collagen matrix in bone repair. Methods Thirty-two adult white New Zealand rabbits were randomly divided into Group Ⅰ and Group Ⅱ. The bone marrow was extracted from tubercle and greater trochanter of femur for culture. Then, the cells were amplified, implanted into type-Ⅰ collagen matrix and further cultured for two weeks. Two weeks later, the cells were implanted into the 1.5 cm radius defect. In the Group Ⅰ, management A (BMS-C+type-Ⅰ collagen) was given to the defect of one radius and management B (control) to the defect of the contralateral radius. In the Group Ⅱ, the defect of one radius was given management A and the defect of the contralateral radius management C (type-Ⅰ collagen).The rabbits were killed 8 and 12 weeks after implantation, the effect of three managements was compared. Results Twelve weeks after implantation, in the management A, all specimens had complete bony union; and in the management B, the specimens were filled with fibrous tissues; in the management C, a little scar formed. Conclusions BMS-C absorbed in the type-Ⅰ collagen can effectively repair bone defect.

Objective To compare the influence of autologous and allogenic bone marrow stromal cells for repairing meniscal injury in avascular zone. Methods Forty New Zealand white rabbits were evenly divided into group A and B. The bone marrow stromal cells from group A rabbits were cultured and mixed with fibrin glue, forming graft. In group A, the graft was implanted into the meniscal injury zone in one side of the knees(autograft group), and only fibrin glue in another side(fibrin glue group). In group B, the graft was implanted into one side of knees (allograft group), and nothing was implanted into contralateral knees(control group). The rabbits were sacrificed 1, 2 and 3 months after surgery, and morphological study was done. Results In the autograft group, fibrous tissue containing a great deal of fibroblast appeared in the injured zone after 1 month, a large number of chondrocytes with formation of collagens were seen after 2 months, and the injuries healed up with fibrous cartilage after 3 months. In the control group, no healing was seen until 3 months. In the fibrin glue group, fibrous tissue was seen with a few fibroblasts, forming scar healing, but no chondrocyte. In the allograft group, the morphological appearance was similar with the autograft group. However, a large number of lymphocytes were seen in reparative tissue. Conclusion Transplantation of bone marrow stromal cells can promote the healing of meniscal injury in avascular zone. Though the allograft of bone marrow stromal cells may induce rejection, its occurrence rate is lower. The allograft for repairing the meniscal injury is worthy of further study.

Objective To evaluate graft isometry after anatomic single bundle reconstruction of anterior cruciate ligament (ACL) using dynamic computed tomography (CT).Methods A retrospective study was conducted of the 14 patients who had undergone single bundle ACL reconstruction from June to August 2015.They were all men,with an average age of 28.6 years (range,from 18 to 39 years).At 6 months after operation,they received dynamic CT scanning during a cycle of knee extension to flexion.3D bone models representing the knee at different flexion positions (0°,30°,60°,90°,and 120°) in each patient were reconstructed from the CT images.The grid method was used to locate the positions of the central footprints of the tibial and femoral tunnels.The lengths between the entries of the femoral and tibial tunnels were measured from each tunnel entry to reflect the graft length change.Furthermore,we measured the isometry at the over-the-top position of the femur and at the anatomic tibial position.Results All the tunnel entries were located at the central area of the ACL anatomic attachment.The reconstructed ACL was the longest when the knee was in full extension.The length was gradually shortened between the femoral and tibial tunnels during flexion of the knee from 0° to 90°.The anatomic position showed an average of 4.82 mm shortening and the over-the-top position an average of 3.28 mm shortening.The length excursion increased in early flexion from 0° to 30° (2.91 ±0.91 mm on average) and reduced in later flexion from 90° to 120° (2.98 ± 1.41 mm on average).Conclusions None of the reconstructed ACL was isometric.A graft length may be the longest when the knee is in full extension and decrease gradually during the flexion from 0° to 90° and increase gradually during the flexion of 90° to 120°.The graft should be fixed when the knee is in the flexion of 30°.

[Objective]To determine the incidence and factors associated with pyrexia after total knee replacement(TKR).[Method]A retrospective review of 200 patients in departments of orthopaedics of our hospital and West China hospital who underwent TKRs from August 1998 and April 2006 was performed.The clinical,surgical and follow-up data were abstracted from the patients' records and were analyzed with SPSS 12.0.[Result]There was a statistically significant increase of the mean temperature post-operations,which remained significant throughout the first 7 days after surgeries(P

[Objective]To study the effects of AO double plates in internal fixation of humeral intercondylar fractures and to analyze the causes of the disfunction in elbow.[Method]From March 2001 to March 2006,21 cases (male 14, female 7) of humeral intercondylar fractures were reviewed, with an average age of 35.3 years (ranged,15～64 years).Left elbow:9 cases;right elbow:12 cases.According to AO/ASIF classification,type C1∶6,type C2∶10, and type C3∶5,2 cases of all were open fractures. Posterior operating approach of elbow: trans-olecranon osteotomy or trans-triceps-side approach and internal fixation by standard method of AO double plates were performed in all cases.The patients began the active training of elbow joint as soon as possible postoperatively.[Result]Twenty-one cases were followed up for 8～38 months and the average time was 14.8 months.Bone union was obtained in all cases.One case complicated wound infection,it recovered after 3 weeks of dressing.And 1 case complicated with radial nerve injury,which recovered with neural nutrients.According to the Cassebaum scoring system,the effects were evaluated as excellent 7,good 10,and fair 4,and the excellent-good rate was 80.9%.[Conclusion]The application of AO double plates was proved to be a good method to treat humeral intercondylar fractures at present. But the severity of the bone comminution and soft-tissue lesion, the effective fixation,earlier active exercise were the main causes that influence the functional recovery of elbow joint.

[Objective] To evaluate the cellular biocompatibility of the nano poly(L-lactic acid)-block-poly(?-caprolactone)(Nano-PLLA-b-PCL)with canine articular cells and its feasibility as a scaffold for the cartilage tissue engineering.[Methods]Nano-PLLA-b-PCL was made by liquid-liquid phase separation.Canine articular cells were isolated and multiplied in vitro.The passage 3 cells were seeded onto the PLLA-b-PCL films and cultured in the 2-dimensional environment.The cytotoxicity was measured with MTT assay.Cellular Morphological changes were observed by phase-contrast microscopy and Hoechst33342 fluorometric method.Another passage 3 cells were seeded onto the Nano-PLLA-b-PCL scaffolds(experiment group),PLLA-b-PCL scaffolds(control group)and cultured in the 3-dimensional environment for 3 weeks.The ratio of cell adhesion was detected by cell counting method.The morphological changes of cells were observed by scanning electron microscopy.The protein content in seeded cells were determined by bioinchoninic acid assay(BCA).The content of DNA was quantified using Hoechst33258 assay.[Results]MTT assay showed the PLLA-b-PCL had no cytotoxicity.The seeded cells adhered and proliferated well into the Nano-PLLA-b-PCL scaffolds,and they maintained good cell phenotype.After 21-day cell culture within the Nano-PLLA-b-PCL scaffolds,the chondrocyte DNA and protein contents increased with time.Moreover,the content of DNA and protein was higher in the experiment group than that in the control group,respectively(P

BACKGROUND: The meniscal allograft transplantation has been proved good outcome in clinical, however, the source and matching restrict its application. By means of a patent technique of separating antigen, a new biological meniscus can be obtained.OBJECTIVE: To observe the histological changes of transplanted biological meniscus in goat's knee joint, in addition, to investigate the feasibility of reconstituting meniscus by using biological meniscus. DESIGN, TIME AND SETTING: The animal experiment of goats was performed at the Central Laboratory in the Third Affiliated Hospital of Sun Yat-sen University between June 2005 and June 2006.MATEAIALS: A total of 15 healthy goats were used in the experiment. Fresh porcine meniscus was harvested for the preparing of biological meniscus.METHODS: The medial meniscus of goat was excised, and then the absent part was repaired using biological meniscus. MAIN OUTCOME MEASURES: The animals were sacrificed at months 1, 3, 6 and 12 after operation. And the samples were detected by histology, scanning electron microscopy (SEM) and Indian ink perfusion. RESULTS: The incision of all animals healed well, without infection or death. The transplanted meniscus healed well with the surrounding tissue, and the connection was tight. With time prolonged, the transplanted meniscus slowly soaked, and live cells existed inside the meniscus, but there were no live cells in about 1/4 district at 1 year after the surgery. Indian ink perfusion showed that the surrounding tissue grew inside and set up microcirculation, but only limited to 10% periphery of the meniscus (normal goat approximately 15%). The transplanted meniscus began the necrosis at 6 months after surgery. Under SEM, the surface of the meniscus became crude and cracked in some places, but still maintains the gross shape at 1 year after the surgery.CONCLUSION: There is no obvious rejection after the biological meniscus was transplanted into knee joint of goats. During the process of absorption, autologous tissue of goats can grow into and set up microcirculation in the transplanted meniscus. The transplanted meniscus protects the'knee cartilage notably, only slightly damage in cartilage can be observed.

BACKGROUND:Bone marrow stromal stem cells(BMSCs)do not allow for single differentiation of chondrocytes due to their multi-directional differentiation,bone morphogenetic protein secreted from osteoblasts affect the non-differentiated precursor cells and promote their osteoblast differentiations,while those differentiated cells are bound to form tissues.OBJECTIVE:To in vitro induce canine BMSCs differentiate into chondrocytes,and to investigate the method and conditions of chondrocyte differentiation in vitro.DESIGN,TIME AND SETTING:Single sample observation was performed in the Laboratory of Tissue Engineering,Sun Yat-sen University between March 2005 and January 2006.MATERIALS:One male dog,aged 4 months,was involved to harvest BMSCs from the rib.METHODS:Rib BMSCs extracted from bone marrow of 2.0-3.0 mL were cultured in vitro. When cells reached a confluence at 8-11 days,trypsinization was conducted and then halted with L-DMEM synthesis culture solution containing 10% fetal bovine serum. Cellular suspension was collected and centrifuged,cells were rssuspended and incubated at a ratio of 1:3. The third generation of cells were cultured and amplified,10 μg/L basic fibroblast growth factor 2 mL was added to replenish culture medium twice,then 1 mg/L transforming growth factor β1 of 2 mL was applied to induce BMSCs differentiation into chondrocytes.MAIN OUTCOME MEASURES:Toluidine blue and alcian blue stains were applied to determine cartilage matdx secretion,immunohistochemistTy was used for the detection of cartilage specific Ⅱ collagen expression.RESULTS:After BMSCs were primarily cultured and subcultured in vitro,they were shown to grow well at the fourth generation,those induced by basic fibroblast growth factor and transforming growth factor β1 were positive for toluidine blue and aician blue staining;immunohistochemistry showed a positive outcome for type Ⅱ collagen,indicating the induced BMSCs exhibited chondrocyte's characteristics.CONCLUSION:Utilizing basic fibroblast growth factor and transforming growth factor β1,the induced canine BMSCs could differentiate into chondrocytes,which is considered as an ideal seed cells for cartilage tissue engineering.

Objective To explore the feasibility of building tissue engineered cartilage by bone marrow stromal cells and pbotografting modified copolymers of 3-hydroxybutymte and 3-hydroxyvalerate.Methods Sheep BMSCs were seeded in three-dimensional photografting modified PHBV scaffoids.Twenty-four hours later.composites were cultured with ehondrogenically inductive medium(DMEM)containing TGF-B(10 ng/m1),IGF-1(150 ng/m1)and 20% fetal bovine serum.Three weeks later,the constructs were evaluated by scanning electron microscopy(SEM)and light microscopy with alcian blue,safrine 0 and type Ⅱ collage immunohistochemical staining.GAG contents of constructs were determined by DMB(1,9-dimethylmethylene blue)binding assay at weekly intervals up to 3 weeks.The composites were implanted subcutaneously in sheep abedoml and were evaluated macroscopically and bistologically at 4 weeks postoperatively.Results SEM photograph showed.after one week culture,cell morphology changed from fibroblast-like elongated spindle to the flat rounded like chondrocytes,and the extra cellular matrix also increased obviousl~.Furthmore,with the culture time extension,this change were more evident.HE staining showed that cells filled all the inter-connected pores in the constructs.And more cells were observed in the outer layer of the constructs.ECM(extraeellular matrix)Was strongly positive by Aleian blue,Safrine O staining and type Ⅱ collage immunohistechemical staining.DMB binding assay revealed that the induced BMSCs GAG secretion(1306.7±192.3)wag significantly higher than BMSCs(205.0±26.2)(P<0.001),but it was significantly lower than passage 2 ehondrocytes(1969.2±235.3)(P<0.001).Saltine O and type Ⅱ collage immunohistochemical staining were positive in constructs implanted subcutaneously.Conclusion Tissue engineered cartilage could be obtained using BMSCs and photografting modified PHBV,but there are still gaps physiologically between the constructs and the nature cartilage.

Objective To explore transinfection of rabbit chondrocytes via chitosan microsphere with human IL-1Ra and TGF-β1 gene. Methods Chitosan-DNA microspheres carrying plasmids with IL-1 Ra and TGF-β1 genes were prepared.The encapsulation efficiency,DNA-released kinetics and lysozyme degradation in vitro were performed.Articular rabbit chondrocytes were co-transinfected with the plasmids with IL-1Ra and TGF-β1 genes via chitosan-DNA mierosphere,evaluated by fluorescence microscope,TaqMan real-time PCR assay and MTF test. Results The chitosan microspheres with IL-1Ra and TGF-β1 genes were(2.8±0.2)μm and(2.6±0.1)μm in diameters respectively.The encapsulation efficiency were(88.3±4.1)%and(87.2±2.6)%.During the degradation,significant morphological changes were noticed.The plasmids could be released in a multiphasic fashion.Enhanced green fluorescent protein and Real-Time PCR analysis showed that genes were expressed in chondrocytes.lasting near 30 days.MTT indicated that the cotransinfection promoted the chondrocytes'proliferation. Conclusion IL-1Ra and TGF-β1 genes cotransfected into chondrocytes via chitosan-DNA microsphere could be expressed in a long term and cotransinfection promoted the chondroeytes'proliferation,which is the base of inhibiting the degeneration of cartilage and promote cartilage repair.