Objective To test the hypothesis that marrow stromal cells （MSCs）, when implanted into selfmyocardium in rabbits, can undergo milieu-dependent differentiation and express cardiomyogenic phenotypes and enhance cardiac function of ischemic hearts, through establish a clinically relevant model for autologous MSCs transplantation, Methods Thirteen New Zealand White rabbits were randomly divided into experimental group （n= 7） and control group （n= 6）. In experimental group, autotogous MSCs（3× 106 cells/30μl） labeled with Bromodeoxyuridine （BrdU） were respectively injected into superior, central and inferior sites in the periphery of the myocardial infarct region. Phosphate buffer saline （PBS） was injected into the scar of the control group hearts according to the same procedure used in the experimental group. Four weeks later, the transplanted labeled MSCs were detected by laser scanning confocal microscopy and the cardiac function were examined by echocardiogram and muhichannel physiologic recorder. Results After 4 weeks, transplanted MSCs were demonstrated myogenic differentiation with the expression of α-sarcomeric actin and connexin 43 located in intercalated disk. MSCs increased the number of vessels compared with controls in myocardial ischemia area. MSCs implantation resulted in markedly improved left ventricular contractility[left ventricular ejection fraction （LVEF）： 0. 51 ± 0.07 vs. 0. 43 ± 0.06 ,left ventricular lateral wall motion distance （LVLWMD） ：1. 75±0. 42mm vs. 1.09±0. 28mm, left ventricular systolic wall thickening ratio（LVAT） ：0. 19%±0.05% vs. 0. 11%±0.04%, left ventricular systolic pressure （LVSP）： 113. 1± 6.3mmHg vs. 99, 5 ± 5, lmmHg, left ventricular end diastolic pressure （LVEDP）： 11. 5±2. lmmHg vs, 14, 3 ±3. lmmHg, maximum rate of left ventricular pressure rise （＋dp/dtmax）：4 618. 3±365. 2 mmHg/s vs. 3 268. 1± 436.9 mmHg/s, maximum rate of left ventricular pressure fall （-dp/dtmax） ：3 008.8±346.7 mmHg/s vs. 2 536.9± 380.4 mmHg/s, P〈0.05]. Conclusion Transplanted autologous MSCs are able to undergo differentiation to form myocardial cells and improve the cardiac function of ischemia myocardium effectively. Autologous MSCs transplantation may have significant clinical potential in treatment myocardial ischemia.
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 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.
OBJECTIVE To explore the healing mechanism of full-thickness wound treating by the intermingled skin transplantation of large sheet allograft with autograft through studying the expression of laminin (LN). METHODS Thirty-six SD rats with 10% to 15% of total body surface area (TBSA) full-thickness were made. After 3 days, the devitalized tissue were excised and transplanted a large sheet of allograft from Wistar rats and islets of autografts were implanted 3 days later. On day 3, 5, 7, 14, 21 after allografting, the expression of LN in the grafts were detected by immunohistochemistry. RESULTS On the 7th day postallografting, LN, which played positive action of epidermal cell adhesion, still retained in the allodermis after the rejection of alloepidermis occurred. On the 14th day postallografting, there appeared scattered LN underneath the epidermal cells migrating from islets of autografts. On the 21st day postallografting, LN in the basement membrane of skin grafts had completely formed. CONCLUSION The intermingled transplantation of large sheet allograft with autograft may provide components of basement membrane for wound healing, which may help to improve the appearance and function of skin.
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 investigate the extent intravenously transplantation of mesenchymal stem cells (MSCs) mediated by magnetic targeting material arrive in the myocardial infarction region and its effects on the recovery of myocardial infarction. Methods Identify the phenotype of the fourth genet of ex vivo expanded MSCs, stain with DAPI after inducing with 10μmol/L 5-aza, then preserve the MSCs for transplantation. 28 SD rats were divided into three groups: group A (n=10), delivered MSCs combined with magnetic targeting material for 30 minutes to rats through tail vein,and kept on raising after placing magnets on the corresponding skin region to myocardial infarction area for 30min; group B (n=9), administration MSCs not conjuncted with magnetic targeting material through tail vein; group C (n=9), direct intramyocardial transplantation of MSCs. Two days after transplantation, evaluate the aggregation state of MSCs in the area of myocardial infarction; 30d later, estimate the functional and morphological changes in myocardial infarction region. Results We observed that each MSCs had 3-5 molecules of magnetic targeting material attached to its membrane under transmission electron microscope. The homing rates of MSCs respectively were group A 38%, group B 6%, group C 53%.The number of aggregating MSCs of group A and group C was apparently more than that of group B（Plt;0.01）. After transplantation, the contraction indices of left ventricle in group A and group C had significant improvement as compared with that of pretransplantation (LVEF 46%±6% vs. 38%±8%, 51%±5% vs. 35%±4%; LVFS 28%±6% vs. 20%±7%, 32%±4% vs. 20%±5%, Plt;0.05) and administrated cells stained with DAPI could be detected in infarction region under optical microscope. After transplantation, the contraction indices of left ventricle in group B hadn’t conspicuous improvement, and the transplanted cells labeled with DAPI could not be identified in infarction region under optical microscope (homing rate of MSCs 38%). There was no statistically difference of results between group A and group C, but in experiment process, there was a high mortality in group C. Conclusion The method that intravenously delivery of MSCs mediated by magnetic targeting material could accumulate much more MSCs in infarction region, reduce infarction size, and effectively improve the cardiac function after infarction.
OBJECTIVE To investigate the mechanism of xenotransplantation rejection and the interaction between the immunocytes. METHODS: This review concluded the research achievements and new advances in xenotransplantation based on the relevant experimental data. RESULTS: Transgenic pig technology and novel immunosuppressants were applied to suppress hyperacute rejection and acute vascular rejection respectively. Modulation of T cell and antigen presenting cells and induction of tolerance were taken for the prevention of acute cellular rejection. CONCLUSION: In general, the technology of transgenic pig is relatively mature and effective. The mechanism and prevention of acute vascular rejection and acute cellular rejection should be further investigated.
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 observe the effect of autologous peripheral blood stem cells(PBSC) transplantation in the treatment of ischemic lower extremity disorders. Methods Therapeutic group:fortyfive patients received recombinant human granulocyte colony-stimulating factor 450 to 600 U/d by hypodermic injection for 5 days to mobilize stem cells.On the 6th day,PBSC were collected by COBE 6.1 Spectra Version and were injected into ischemic lower extremity. Control group:33 patients were treated with dilating vessels drugs. After operation some indexes were evaluated. Results After operation, these indexes were improved. Skin temperature and TcpO2 were improved obviously, being statistically significant difference(P＜0.05). Conclusion Autologous PBSC transplantation might be a safe and effective method for treating lower extremity ischemic disorder. It could improve the quality of life of many patients as amputation of lower extremity of foot might be avoided or reduced.
Objective To establish a rat model of pancreas-duodenal transplantation for pancreas transplantation research. Methods A rat model of pancreas-duodenal transplantation was established by using dual cuff technique. The graft affiliated portal vein and abdominal aorta were anastomosed to recipient’s left renal vein and left renal artery by using dual cuff technique without clamping the systemic circulation in order to minimize hemodynamic instability. Simultaneously the graft duodenum was anastomosed to the host jejunum in an end-to-side fashion to reestablish drainage of pancreatic secretion. Fluid replacement, warm keeping and anticoagulation were maintained during perioperative period. Results The average donor operation time was (68.4±7.2) min and recipient operation time was (26.1±3.3) min. Moreover, it took (5.0±1.1) min for cuff preparation in vitro and (9.6±3.5) min for vessel reconstruction in vivo, respectively. Intestinal anastomosis took (7.2±2.3) min. The operation successful rate was about 91.7%. Conclusion This pancreas-duodenal transplantation model avoids systemic circulation clamping. It is simple and stable, with the value in pancreas transplantation research.