Objective To study the effect of transforming growth factor β1 (TGF-β1) plasmid on poly frosted-defrosted allogenic nerve transplantation. Methods Forty Wistar rats were randomly divided into two groups equally. A 2.0 cm sciatic nerve segment, 5 mm away from infrapiriformis muscle space, was removed and the defect was repaired with poly frosteddefrosted allogenic nerve. The TGF-β1 plasmids were injected into the nerve anastomosis and adjacent muscles in the experimental group, normal saline in the control group. The nerve specimens were sectioned for staining in the 6th and 12th weeks . Axonal count and statistical analyses were done. Results The grafted and distal nerve segments showed regenerated fibers in both groups. In the experimental group,less edema and more nerve fibers were observed in the 6th week. The grafted nerve segment was filled with regeneration axons, the myelinated nerve fibers arranged regularly, and the axons and the myelin sheaths developed well in the 12th week. There was significant difference in the number of regenerating axons between the experimental group 98.6±4.8／μm2 and control group 75.8±5.1／μm2 (Plt;0.01). Conclusion Multiple frost-defrost of allogenic nerve can reduce its antigenicity and increase itsusefulness in repairing nerve defects. Local use of TGF-β1 plasmid can enhance immunosuppression to reduce immuno rejection.
Objective To study the feasibility of transplanting human saphanous vein endothelial cells to luminal surface of blood vessel prosthesis and to play a theoretical foundation for the clinical application of autologous endothelial cell transplantation. Methods Human saphanous vein endothelial cells were harvested with 0.1% collagenase and cultivated in vitro for 13.08±1.24 days. The cultures were confirmed as endothelial cells with the fourescent linked anti-Ⅷ antigen antibodies. The content of both 6-keto-PGF1α and Von Willebrand factor (vWF) in the supernatant were detected with ELISA and radioimmunoassay. The multiplied cells were lined in vitro onto the luminal surface of expanded polytetraflouroethylene (ePTFE) grafts precoated with fibrin glue and fibronectin, then cultivated again for 9 days. Results 11.46±2.69×１０６ of available endothelial cells could be regularly obtained, the number of endothelial cells increased 147.93±88.68 times when culture were terminated. All the cells diploid cells with a purity of 99%. The content of both 6-keto-PGF1α and vWF in the media showed no significant difference between the primary and subculture passages. The luminal surface of grafts was covered completely by a spindlelike endothelial monolayer and an even fibrin glue matrix could be seen underneath. Conclusion Endothelial cells derived from human saphanous veins might be feasible to be transplanted onto the luminal surface of ePTFE and present a potential clinical application.
Objective To establish a scaffold model from heterogeneoussmall blood vessels. Methods Caudal arteries from 34 Wistar rats( average length 12.08±1.69 cm) were made into acellular blood vessel scaffolds. Some scaffoldswere observed by electron microscope, and others were transplanted to the cut ends of ear central arteries of male Japanese big ear white rabbits. Results Average external diameter was 0.74±0.08 mm in proximal, and 0.55±0.08 mm in distal end of rat caudal arteries. The small blood vessel scaffolds had shin wall whichwas white and soft, composed of fibrous tissues without cells. On the intima surface the fibrous tissues were arrayed densely in a grid-like pattern. After transplantation, the blood flow was reserved, and kept flowing freely in 24 hours. The pulsation of the transplanted artery was accessible and no blood leakage wasfound.Conclusion The natural scaffolds are composed of fibrous tissues, and can sustain the artery pulse pressure for 24 hours. It is better to suture the blood vessels by sleeve anastomosis.
Objective To explore the methods of repairing cartilagedefects and to introduce the clinical experience with the autologous osteochondral transplantation. Methods Twenty-five patients with chondral and osteochondral defects of the weight-bearing surfaces were treated by the autologous osteochondral transplantation for the repair of the chondral and osteochondral defects of the unweightbearing surfaces under arthroscope. According to the shape of the defects, the different dimensions of the osteochondral autograft were selected. All the patients began the training of the continuous passive motion after operation. Six weeks after operation, the patients began to walk in the weightbearing habitus. However, in the control group, another 25 patients were retrospectively analyzed, who had chondral and osteochondral defects of the weight-bearing surfaces but were treated only by the cleaning and drilling procedures. The scores evaluated bythe Brittberg-Peterson scoring scale of the 2 group were 98.65±9.87 and 96.98±8.94 respectively. Results The follow-upfor 3-24 months after operation revealed that the treated knee joint had a goodmotion extent. The pain was obviously alleviated. Based on the longitudinal study with the three-dimensional spoiled magnetic resonance imaging (MRI), the signal intensity of the repaired tissues approached to the normal condition. The scores evaluated by the Brittberg-Peterson scoring scale were almost zero 3 monthsafter operation in the experimental group, and the scores were 58.48±6.98 inthe control group. There were significant differences between the experimental group and the control group(P＜0.01). Conclusion Autologous osteochondral transplanation under arthroscope is a good curative method for the cartilage defects, with advantages of minimal invasiveness and avoidanceofrejections resulting from allografts. However, its long-term effect needs to befurther studied. The conventional therapies including cleaning and drilling are useful in alleviating the symptoms.
Objective To design a combined flap of subscapular axis including vascularized lateral scapular,rib and latissimus dorsi to repair the large defect of tibia. Methods The patient was a 39-year-old man who got a posttraumatic 12 cm defect of tibiaafter primary debridement and external fixation because of open fracture 5 months ago. There was a 12 cm×6 cm scar involved the proximal medial segment of tibia.After resection of scar and fibular tissue over the bone defect floor, alatissimus dorsi myocutaneous flap 14 cm×5 cm pedicled with subscapular artery-thoracodorsal artery,a flap 12.5 cm on the outside of the scapular pedicled with thoracodorsal artery, and 6th rib flap 13 cm by serratus were prepared.The tibialis posterior and saphenous vein were used for astomosis. A proximalanatomic plate was applied to the fixation of tibia. Results Thecompound flap survived the operation. The follow-up period was 2 years. Bone union occurred 6 months after operation. Conclusion This combined flap is successful and can provide alternative to the resolution of large defect of tibia.
OBJECTIVE: To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. METHODS: Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2, 4, 8 and 12 weeks after transplantation. RESULTS: There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4, 8 and 12 weeks after transplantation (P lt; 0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. CONCLUSION: It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute.
Objective To summarize the advancement of immune tolerance in pancreas transplantation.Methods Relevant literatures about immune tolerance in pancreas transplantation, which were published recently domestic and abroad were collected and reviewed. Results The main methods to induce immune tolerance are peripheral tolerance and central tolerance. The induction of chimerism by infusion of donor-specific bone marrow cells is the research hot spot recently. Conclusion The infusion of donor-specific bone marrow cells in combination with one or more peripheral tolerance maybe can induce immune tolerance successfully. However, it should be researched further.
Objective To evaluate the biocompatibility of a new bone matrix material (NBM) composed of both organic and inorganic materials for bone tissue engineering. Methods Osteoblasts combined with NBM in vitro were cultured. The morphological characteristics was observed; cell proliferation, protein content and basic alkaline phosphatase(ALP) activity were measured. NBM combined with osteoblasts were implanted into the skeletal muscles of rabbits and the osteogenic potential of NBM was evaluated through contraat microscope, scanning electromicroscope and histological examination. In vitro osteoblasts could attach and proliferate well in the NBM, secreting lots of extracellular matrix; NBM did not cause the inhibition of proliferation and ALP activity of osteoblasts. While in vivo experiment of the NBM with osteoblasts showed that a large number of lymphacytes and phagocytes invading into the inner of the material in the rabbit skeletalmuscle were seen after 4 weeks of implantation and that no new bone formation was observed after 8 weeks. Conclusion This biocompat ibility difference between in vitro and in vivo may be due to the immunogenity of NBM which causes cellular immuno reaction so as to destroy the osteogenic environment. The immunoreaction between the host and the organic-inorganic composite materials in tissue engineering should be paid more attention to.
Tissue engineering trachea is an artificial trachea with biological activity, which is constructed in vitro by using tissue engineered principle and technology, and is a tracheal prosthesis for replacing large circumferential defect of the trachea. The course of its construction is as follows. First, seeding cells are cultured and expanded in vitro. Then they are collected, counted and seeded onto the biomaterial scaffold of tissue consistent and biodegradation. Finally, the biomaterial-cells construction is implanted into bio-reaction device or one’s subcutaneous layer. The tissue engineering trachea could be constructed after cultured certain times. Compared with other artificial trachea, the tissue engineering trachea has more advantages, such as nonimmunogenicity, no side-effects related to foreign graft materials, and biologic activity. This will bring some hope to look for an appropriate graft material. However, the study about it is still faced with some difficult problems, such as vascularized trachea, culturing in vitro, and prevention of infection in trachea prosthesia. So there will be long time for tissue engineering trachea to apply clinical tracheal transplantation successfully. This assay has reviewed the study about tissue engineering trachea from three sides——the source of seeding cells, the research about biomaterial scaffold, and the construction of tissue engineering trachea.
OBJECTIVE To prevent early closure of growth plate and developmental deformities of limbs by allografts of cultured cartilages into growth plate defects of rabbits. METHODS Chondrocytes isolated from articular cartilage of 1-month rabbits formed cartilage after cultivation in centrifuge tubes. The cartilages cultured for two weeks were implanted into growth plate defects of proximal tibiae of 6-weeks rabbits. At 4th and 16th weeks, X-ray, histologic and immunohistochemical examination were performed. RESULTS The tibiae had no marked deformities after 4 weeks of operation. Histologic examinations showed that the defects were filled with cartilage. Immunohistochemical results of type II collagen were positive. The tibiae with allografts of cultured cartilages had no evident deformities after 16 weeks of operation. Histologic examination showed nearly closure of growth plates. On the contrary, the tibiae on control side formed severe deformities and growth plate were closed. CONCLUSION Allograft of cultured cartilages into growth plate defects may replace lost growth plate tissues, maintain normal growth of limbs and prevent developmental deformity.