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.
In order to compare the immunogenecity and biological properties of homologous tendon grafts after treatment from different methods of freezing, tendons from chickens received repeated freezing-thawing treatment or ultra-low-temperature treatment, and then, the post-treatment tendons were preserved in liquid nitrogen for 3 months before transplantation. The autogenous tendon transplantation was served as the control. It was found that in the group of repeated freezing-thawing treated tendons, the tendon cells all died and while in the ultra-low temperature treated tendons the active rate of tendon cells was 92.5% +/- 3.4%, and the histological observation showed that transplantation of frozen tendons would result in extensive infiltration of inflammatory cells in the grafted tendons and the peritendinous adhesion was serious than that of the autografts. The active flexion function, hydroxyproline levels and the biomechanical analysis showed no significant differences between the repeated freezing-thawing treated homografts and the ultra-low-temperature treated homografts, and that the autografts was definitely superior to the homografts. The conclusions were: (1) Transplantation of the homologous tendons from the two different methods of freezing could receive considerable success and there was no significant difference between them; (2) Transplantation of frozen homologous tendon graft might give successful result which was probably due to the preservation of the cellular activity of the tendon cells following freezing treatment and elimination of the antigen presenting cells in the tendon as well, and (3) Although the cellular components of the tendon were damaged and the antigenicity of the tendon was lowered, it did not necessarily mean that homologous tendon graft would always be successful in transplantation.
OBJECTIVE:To investigate the index of the rejection of lJle retinal pigment epithelium(RPE)cells transplantation. METHOD:Allogenic RPE transplantation on rahbits by transcleral technique, the changes of interleukin-2 (IL-2) activity in peripheral blood and the effect of immunoinhibitor (methylprednisonlone)were detected. RESLILTS:In the group of simple transplantation,the IL-2 activity in peripheral blood begin to rise in the first day after operation. The peak value occured in the third day,and is still much higher than that of the control group in the 14th day,whereas in the group treated with immunoinhibitor ,there was no obvious difference in the first day after operatlon,in the third day,the IL-2 activity rises slightly,and returned to normal level in the 7th day. CONCLUSION: After RPE transplantation, the level of IL-2 activity in peripheral blood might serve as an important index to determining and detecting the rejective response. （Chin J Ocul Fundus Dis,1996,12: 239-241）
Objective To observe the effect of transplantation of embryonic stem cell(ES) on neurological functional recovery of injured spinal cord in adult mouse. Methods The ES cells were cultured and induced in vitro. Fifty C57/BL6J mice were made animal model of semicut mice of T9，10. The ES cellderived neural precursors cells were transplanted into the vertebral canalaround injured spinal cord semi-cut mice. Twenty-eight C57/BL6J mice were randomly divided into three groups: sham operation group(group A,n=9), operation／cell group (group B，n=10), and operation／DMEM group(group C,n=9). RT-PCR analysis, X-gal staining and immunofluorescence were used to observe the cells survival and differentiation in the spinal crod. BBB test was performed to study functional improvement. Results ES cells induced and cultured in vitro displayed clonal growth with circle or ovoid shape and had one or more nucleoli. RT-PCR result showed that the induced ES cells expressed mRNA of Nestin and microtubuleassociated protein, but did not express glial fibrillory acidic protein(GFAP). There was statistically significant difference in BBB scoring between group A and groups B, C after operation (P＜0.01). There was statistically significant difference in BBB scoring at 1, 2 and 4 weeks of operation(P＜0.01), but no statistically significant difference at 6 and 8 weeks of operation between groups B and C(P＞0.05). The X-gal staining results werepositive in group B and negative in groups A and C. The immunoflurescence resultshowed neurofilament green fluor and no expression of GFAP in injured spinal cord region. Conclusion After transplantation, ES cellderived cells can survive, transfer into the injury position, and differentiate into neurons, but spinal cord function has no obvious improvement.
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 clinical result of treatment of bonecyst by transplantation of the autologous bone marrow combined with the allograft bone. Methods From February 2004 to March 2006, 13 patients withbone cyst were treated by transplantation of the autologous bone marrow combined the the allograft bone. Among the 13 patients, 6 were males and 7 were females, ranging in age from 5 to 16 years, averaged 11.5 years. In the patients, 5 lesions were located inthe proximal humerus, 2 in the femoral neck, 3 in the femoral shaft, 2 in the proximal tibia, and 1 in the distal tibia. Among the patients, 5 had a complication of pathologic fracture. All the patients underwent an erasion of the bone cyst, and then the transplantation of the autologous bone marrow combined with the allograft bone, and 8 of them were also given an instrument fixation. Results The follow-up for 6 months to 2 years after operation revealed that 5 of the patients had an incision healing by the first intention, 5 had an effusion in the incision site, and 3 had a delayed healing of the incision. According to the Capanne criteria, the postoperative X-ray findings indicated that 10 patients had Grade Ⅰ healing, and 3 had Grade Ⅱ healing. The complete healing took 3.5-8 months,averaged 5.2 months. There was no recurrence. When the fixation instrument was removed, no pathologic fracture occurred. The function of the upper and lower limbs recovered. Conclusion Transplantation of the autologous bone marrow combined with the allograft bone is an effective and safe procedure for treatment of bone cyst.
Objective To evaluate the clinical effects of fibula flap grafts on the repair of the extremities with traumatic compound tissue defects. Methods In 12 cases, the fibula flap grafts were employed to restore the extremities with traumatic compound tissue defects. Of the 12 patients, 9 were males, 3 were females; their ages ranged from 12 to 45. There were 2 cases of tibia defect combined with fibula fracture, 2 cases of tibia defect, 2 cases of radius defect, 3 cases of ulna defect, 1 case of calcaneus defect,and 2 cases of firstmetatarsus defect. The bone defect length ranged from 4.2 to 10.6 cm, 7.8 cm in average.The skin defect area ranged from 10.0 cm×4.5 cm to 27.0 cm×15.0 cm. The free transplantation of fibular flaps were used in 9 cases, the lapse operation were used in 2 cases, retrograde shift were used in 1 case. Results Postoperational vein crisis and commonperoneal nerve traction injury were observed in category mentioned above respectively. All the 12 fibula flaps survived after proper treatments such as removalof great saphenous vein. Follow-ups were done for 6 to 24 months. Both the transferred fibula and the recipient broken end reflected bones were healed. Four patients underwent the second-phase reconstruction operation oftendon moving power. One wrist and 1 ankle underwent arthrodesis in 3 to 6 months.All the effects were satisfactory. Conclusion The fibula flap grafts provide arelatively better alternative to repair the extremities with long bone compoundtissue defects. In addition, the sensory function reconstruction of fibula flaps should be given full attention.