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 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 investigate the therapeutic effect of flap transplantation in repairing soft tissue defects of children. METHODS: From January 1997 to May 2002, 75 cases of different soft tissue defects (52 males and 23 females, with the age of 3-14 years) were repaired by axial and non-axial flaps transfer, and axial flaps transplantation by microvascular anastomosis. The flaps area ranged from 3 cm x 5 cm to 15 cm x 42 cm. Emergency operation was performed in 26 cases and secondary operation in 49 cases (infective wound such as osteomyelitis and plate extra-exposed of fracture). The defect regions included the forearm, back of the hand, thumb, index finger, leg and foot. The types of flap graft and application range included 39 cases of axial flaps transfer or transplantation (27 cases of along- or contra-transfer of transplantation and 12 cases of microvascular anastomosis). The non-axial flaps transfer were designed along- or contra-transfer near the wound area in 36 cases. The ratio of length to width was 2.5:1-3.5:1 in 27 cases, and larger than 3.5:1 in 9 cases. Adequate anesthesia method should be chosen according to the characteristics of children, non-traumatic operating during surgery and postoperative supervision and nursing of flaps should also be paid enough attention. RESULTS: After operation, blood circulation crisis occurred in 2 cases (1 case of artery failure and 1 case of vein failure). The flaps survived in 37 cases and partially survived in 1 case and necrosed in 1 case. The survival rate was 96.2%. The postoperative follow-up period was 3 to 60 months, the blood supply, elasticity and texture of flaps were good. The effect of repair was satisfactory. CONCLUSION: Different types of transplantation of blood-supply of flaps may repair the different types of soft tissue defects in children. Free flap transplantation is safe and beneficial in children, different defects of soft tissue were repaired by axial and non-axial flaps transfer, axial flaps transplantation by microvascular anastomosis. Non-traumatic operating and postoperative supervision and nursing of flaps should also be paid enough attention.
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.
In the study of repair of massive bone defect with free vascularized fibula graft, 13 cases were reported, in which traumatic defect in 7 cases, segmental resection of bone from tumors in 5 cases and osteomylitis in 1 cases. They all were treated successfully with vascularized fibular graft. After a follow-up of 6 months to 7 year, bone healing was observed with satisfactory and rehabilitation of functions. In one case, fatigued fracture occured twice due to early walking. It was concluded that free vascularized fibular graft was very helpful in the repair of massive bone defect, but prolonged external fixation after operation might be important to prevent fractur of grafted bone.
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.
【Abstract】ObjectiveTo investigate the effect of xenotransplantation of microencapsulated rabbit parathyroid tissue in different sites in rats for the treatment of hypoparathyroidism. MethodsThe parathyroid glands from Wistar rats were removed to make them aparathyroid. Ultimately, sixteen rats were included because their serum calcium values were continuously below 1.6 mmol/L. We also encapsulated the cultured rabbit parathyroid tissue with alginateBaCl2 microcapsule. According to the transplantation sites, rats were randomly divided into two groups: renal adipose microcapsule group and peritoneal microcapsule group, eight in each group. Encapsulated rabbit parathyroid tissues were then transplanted accordingly to different microcapsule groups. The calcium serum contents were examined on 5，15，25，35，45，55 and 65 d respectively after transplantation and the grafts were observed through electron microscope on the 65 d in particular. ResultsThe calcium contents after transplantation in renal adipose microcapsule group restored to normal and the observation outcomes of grafts showed that they survived well. The calcium contents of posttransplantation in peritoneal group also restored to normal with an exception that it dropped to a level lower than 1.6 mmol/L on the 65 d. Electron microscope also showed that there were necrotic tissues in the center and only a few cells survived on the edge of the grafts. Within peritoneal microcapsule group, the values were significantly lower than others taken at different phases. ConclusionMicroencapsulated rabbit parathyroid tissue that was xenotransplanted into rats can survive and function without administration of immunodepressant. There are significant differences of calcium contents at varying phases between two transplantation sites, which demonstrate that renal adipose may be an optimal site for microcapsule xenotransplantation.
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 review the advances in repair of spinal cord injury by transplantation of marrow mesenchymal stem cells(MSCs). Methods The related articles in recent years were extensively reviewed,the biological characteristic of MSCs,the experimental and clinical studies on repair of spinal cord injury by transplantation of MSCs,the machanisms of immigration and therapy and the problems were discussed and analysed. Results The experimental and clinical studies demonstrated that the great advances was made in repair of spinal cord injury by transplantation of MSCs. After transplantation, MSCs could immigrate to the position of spinal cord injury, and differentiate into nervelike cells and secrete neurotrophic factors.So it could promote repair of injuryed spinal cord and recovery of neurologicalfunction. Conclusion Transplantation of MSCs was one of effective ways in repair of spinal cord injury, but many problems remain to be resolved.
Objective To investigate the myogenic differentiation of mesenchymal stem cells (MSCs) after being transplanted into the local muscle tissues. Methods The serious muscleinjured model was established by the way of radiation injury, incising, and freezing injury in 36 mouses. Purified MSCs derived from bone marrow of male mouse and MSCs induced by5-azacytidine(5-Aza-CR) were transplanted into the local of normal muscle tissues and injured muscle tissues of femal mouse. The quantity of MSCs and the myogenic differentiation of implanted MSCs were detected by the method of double labeling, which included fluorescence in situ DNA hybridization (FISH) and immuno-histochemistry on the 1st, 3rd, 6th, 9th, 12th, and 15th day after transplantation. Results The quantity of implanted MSCs decreased as timepassed. MSCs’ differentiation into myoblasts and positive expression of desmin were observed on the 15th day in purified MSCs group and on the 6th day in induced MSCs groups. Conclusion MSCs could differentiate into myoblasts after being implanted into the local of muscle tissues. The differentiationoccurs earlier in the induced MSCs group than that in purified MSCs group.
