We have performed 27 cases of orthotopic homologous cardiac transplantation using Korean mongrel dogs and one case of sham operation for the evaluation of harmful effect of cardiopulmonary bypass itself on the dog from April, 1989 to June, 1990. Our previous reports have already demonstrated basal hemodynamic and hematologic data on the canine homologous heart transplantation and the fundamental principles of transplantation of the heart. The mean body weight of recipients was 13.2$\pm$1.2kg with a rage of 11 ~ 15kg, and the hemodynamic and hematologic pictures were almost same as the result of previous reports from our hospital, except marked decrease in postoperative platelet count[from 3.18 $\pm$0.80x106/mm3 to 1.41$\pm$0 37x 106/mm3]. Mean survival time was 24.82$\pm$49.40 hours with the longest survival of 264 hours. Donor cardiectomy included coronary vasodilatation with diltiazem, potassium arrest, and the rapid cooling of the heart suspending in the specially designed ice-bath. Median sternotomy provided excellent exposure of the surgical field. 6 \ulcorner0 prolene suture was used for the anastomosis of both atrial cuffs and the great arteries, and we found the fact that stenosis, bleeding, thrombus formation around the anastomotic site could be decreased with the use of everted horizontal mattress suture techniques. Immunosuppression was done with a combination of lower dose Cyclosporin-A, Azathioprine, methyl-prednisolone, but our cases still showed too short survival to worry about graft rejection. Still poor was our quality control of experimental animal, we had much difficulties in postmortem evaluation of the dogs. Low cardiac output due to biventricular failure, intractable supraventricular or ventricular tachyarrhythmia, postoperative massive bleeding, sepsis were most frequent findings that could be thought as a cause of death. A few cases showed subendocardial patch hemorrhage in both ventricular cavity or atrial septum at autopsy, suggesting acute subendocardial infarction. Although our team overcome most of the technical problems of orthotopic heart transplantation, we should pile up further knowledges about donor heart preservation, quality control of animal, infection, rejection, the effect of the cardiopulmonary bypass to improve the results.
The intact dental pulps which were free of their tooth bud from adult rat incisors, and oral mucosa were transplanted subcutaneously in homologous rats to study the formation of calcified tissue. The rat were sacrificed after 1,2,3 and 4 weeks following transplantation of dental pulp and oral mucosa. The samples which contained the transplanted and surrounding tissue were fixed in 10% NBF, stained with hematoxylin and eosin, alizarin red S, von Kossa, and alcian blue. Microscopic examinstins revealed as follows: 1. The transplanted oral mucosas were not calcified but tended to form the epithelial cysts. 2. At 1 week after transplantation of dental pulp the calcified structures were appeared at the periphery of the transplantation of dental pulp but weakly reacted to alizarin red S, von Kossa, and alcian blue. 3. At 2 weeks after transplantation of dental pulp the calcified structures began to expand from the periphery to the center of the transplanted dental pulp and occupied the large areas comparatively, and strongly reacted to alizarin red S, and von Kossa stains. 4. At 3 weeks after transplantation of pulp tissue the fibrous components were grown at the periphery of the transplanted pulp tissuesand at 4 weeks a large amount of fibrous tissues were observed. The transplanted pulp tissue tended to form foreign bodies gradually.
Background: The replacement of the narrowed long-segment trachea with various prosthetic materials or tissue grafts remains a difficult and unsolved surgical problem. Homologous cryopreserved tracheal transplantation has been considered to treat the irreversibly-damaged organs, such as in the lung or heart transplantation and also to overcome the limited supply of donor organs. We examined the morphological changes and the immunosuppressive effects of the cryopreserved trachea after the heterotopic transplantation in the rats. Material and Method: Sixty tracheal segments harvested from 30 donor Wistar rats were heterotopically implanted into the peritoneal cavity of 20 recipient Wistar rats and 40 Sprague Dawley rats. The 60 recipient rats were divided into 6 groups(10 rats/ group). In groups I, II, and III, 30 tracheal segments were implanted immediately after the harvesting and in groups IV, V, and VI, the segments were implanted 28 days after the cryopreservation. Groups I and IV were Wistar syngeneic controls. Groups II and V were Sprague Dawley recipients receiving no immunosuppression and Groups III and VI, were Sprague Dawley recipients receiving immunosuppressive agents. At 28 days all rats were sacrificed and the tracheal segments were evaluated grossly and histologically. Result: Immunosuppression of the tracheal segments had a significant influence on the changes of the tracheal lumen and tracheal epithelial cells, irrespective of the cryopreservation of the trachea(p<0.001). In groups III and VI receiving immunosuppressive agents, the tracheal lumen was patent and the normal epithelial cells were observed, however in the other groups not receiving the immunosuppressive agents, there were almost luminal obliteration by the proliferation of the fibrous tissues and a loss of the epithelial cells, the findings were similar to those in the case of obliterative bronchiolitis after a lung and a heart-lung transplantation. Conclusion: With the appropriate immunosuppressive agents, the lumen and the respiratory epithelium of the transplanted tracheal segment were well preserved, even after the cryopreservation of the tracheal segment, which shows the possibility of the long-term preservation and homologous transplantation of the trachea. But fibroproliferative obliteration of the tracheal lumen and the loss of the normal respiratory epithelial cells, characteristic findings of obliterative bronchiolitis, were observed in the groups without the immunosuppression. This experiment using the rat trachea may be useful in studying the pathogenesis, treatment, and prevention of obliterative bronchiolitis after a lung and a heart-lung transplantation.
Heterotopic abdominal homograft of canine heart was carried out in 20 pairs of dogs. Of these 12 cases were subjected as a control and 8 were subjected to immunosuppressive group. The dosage of immunosuppressive agent was 5mg/kg/day of Imuran [Azathioprine] for 3 days preoperatively, 10mg/kg on operative day and 5mg/kg/day postoperatively. For reducing the metabolic demand, the donor heart was preserved in 4degree heparinized saline solution for approximately I4 minutes. In the most of the cases, transplantation was performed with the technique of end-to-side aorto-aortic anastomosis and end-to-side pulmonary artery-inferior vena cava anastomosis at the infrarenal portion. Five out of 20 grafted dogs were survived more than one day. The longest survived 18 days in the control group and survive more than 60 days in the treated group. The survival cases were 3 out of 8[37. 5%] in the group of dogs treated with lmuran and 2`out of 12 [16.6%] in the group of non-treated. A prominent gross findings of the grafted heart was a minimal to moderate degree of dilatation of the heart with or without thrombosis in the cardiac chambers and/or anastomotic site. The case number 10, 15, and 19 showed moderate hypertrophy in grossly. The microscopic findings were as follows; 1. There were early hypersensitive histologic reactions such as interstitial edema, cellular infiltrations and early degenerative changes in the myocardium in the cases of 3 hour survival. 2. In the cases of more than 6 hours survival, organizing thrombosis of myocardial vessels, vasculitis,myocardial necrosis and lymphocyte, plasma cell, round cell infiltrations were noted. In the cases of more than 12 hours survival, the degree of these histologic changes especially in the non-treated group were more intensified than in the treated. 3. In the cases which survived more than one day, so called homograft specific histologic changes were milder in the immunosuppressive group compared with the control. 4. All the host hearts showed no evidence of pathologic findings histologically. Among the homologous canine cardiac transplantation tissue reaction, was milder and suvival time longer in the group treated with immunosuppressive drug.
Purpose: This study was aimed to evaluate the effect of the Freeze Dried Bone Allograft and Demineralized Bone Matrix on osseous regeneration in the rat calvarial defects. Methods: Eight mm critical-sized calvarial defects were created in the 80 male Sprague-Dawley rats. The animals were divided into 4 groups of 20 animals each. The defects were treated with Freeze Dried Bone Allograft($SureOss^{TM}$), Demineralized Bone Matrix($ExFuse^{TM}$ Gel, $ExFuse^{TM}$ Putty), or were left untreated for sham-surgery control and were evaluated by histologic and histomorphometric parameters following a 2 and 8 week healing intervals. Statistical analysis was done between each groups and time intervals with ANOVA and paired t-test. Results: Defect closure, New bone area, Augmented area in the $SureOss^{TM}$, $ExFuse^{TM}$ Gel, $ExFuse^{TM}$ Putty groups were significantly greater than in the sham-surgery control group at each healing interval(P < 0.05). In the New bone area and Defect closure, there were no significant difference between experimental groups. Augmented area in the $ExFuse^{TM}$ Gel, $ExFuse^{TM}$ Putty groups were significantly greater than $SureOss^{TM}$ group at 2weeks(P < 0.05), however there was no significant difference at 8 weeks. Conclusions: All of $SureOss^{TM}$, $ExFuse^{TM}$ Gel, $ExFuse^{TM}$ Putty groups showed significant new bone formation and augmentation in the calvarial defect model.
Background: Recently, open heart surgerys using homograft are progressively increasing in complex cardiac anomalies, and even though the use of homograft tissues harvested from hearts of transplant recipients and brain-death patients are allowed and their use is increasing, the supply of homograft tissue is very limited. Material and Method: The large diameter homografts are difficult to apply directly for RVOT reconstruction of small neonatal and infant hearts due to the size mismatching. Therefore, were surgically down-sized the large diameter tricuspid homograft into bicuspid conduits by means of a longitudinal incision of the oversized homograft, excision of one cusp, and oversewing of the“Bicuspid homograft”wrapped around a Hega dilator of the appropriate size. Result: 3 patients(Male 1, Female 2: tetralogy of Fallot with pulmonary atresia), ranging in age from 5 months to 4 years and ranging in weight from 5.5Kg to 12.95Kg underwent reconstruction of the RVOT with bicuspid conduits obtained by appropriate tailoring from large-diameter homografts. The mean follow-up period was 4.3 months(range, 2 to 6 months). There were no complications related to the homograft tissues. Conclusion: In the short term follow-up, the bicuspid homografts provided good competence and excellent hemodynamics although a long term follow-up is needed to assess the functions of the bicuspid homografts in RVOT. We believe this technique may be a more effective alternative than the use of synthetic conduits when the use of an appropriate-sized homograft is not possible.
In order to test the hypothesis that the pulmonic valve, when used to replace the aortic root as a pulmonary autograft, will remain a viable anatomical structure and will grow and develop normally along with the host, we performed aortic valve replacement with the pulmonary autograft in 15 neonatal piglets. The weight of the donor was 9.3 $\pm$ 0.2 kg, the recipient 9.6 $\pm$ 0.3 kg. Measured diameters of pulmonic annulus were 14 $\pm$ 0.2 mm for autograft and 14.2 $\pm$ 0.2 mm for pulmonary artery homograft. Operation was performed under cardiopulmonary bypass with deep hypothermia [20oC at low flow perfusion [70 ml/kg/min . The mean operation time was 227 $\pm$ 10 min., bypass time 152$\pm$ 7.6 min. and aortic cross clamp time 73$\pm$ 4.6 min.. 9 piglets survived more than 12 hours. One survived 12 days and died of pneumonia and the latest one survived in good condition and sacrificed at postoperative 6th week for cardiac catheterization and pathologic examination that revealed the viability and growing of the pulmonary autograft. Currently we are able to complete the operation with good preservation of cardiac function, and our postoperative care has evolved to the extent that we are now confident enough of having an acceptable percentage of long term survivors to undertake a definite study in this regard.
Allograft cardiac valves have been used for over 30 years to replace diseased cardiac valves, reconstruct right or left ventricular outflow tract. With increasing its requirement, the establishment of a viable bank capable of maintaining the viability of graft over a prolonged period would be desirable. The method for determining the viability of allograft by metabolic assay technique using radiolabeled aminoacids has been used recently. An experimental study was done for evaluation of viability of cardiac allograft which was preserved for 14 days at 4oC in nutrient medium[fresh preservation] by metabolism assay technique using 3H-glycine. Also, the effectiveness of low concentration antibiotic solution[CLPV] for sterilization was evaluated. The effectiveness of CLPV solution for sterilization of allograft was perfect. Pre-treatment cultured organisms were not cultured after treatment at all in every cases. The viability of allograft after sterilization was reduced to 66.4%[aortic wall], 74.7%[pulmonary wall], 76.3%[aortic valve], 67.9%[aortic wall]. And after the fresh preservation for 14 days, the viability was reduced to 14.7%, 18.5%, 17.7%, 19.0%, respectively.In conclusion, viability of allograft was reduce to 71.3[66.4-76.3]% after sterilization and 17.5[14.7-19.0]% after fresh preservation. And sterilization effect of CLPV solution was satisfactory.
Background: There are no ideal substitutes for tracheal replacement. Therefore we investigated the possibility of clinical use of cryopreserved tracheal homograft with special interest in the viability and rejection of the epithelial cell and cartilage. Material and Method: Rabbit's trachea was sected and stored in liquid nitrogen tank for 1 month. Tracheal replacement was done in 45 rabbits with autograft(n=15, Group 1), fresh allograft(n=15, Group 2) and cryopreserved homograft(n=15, Group 3). After 7, 14, and 30 days, 5 rabbits in each group were sacrificed and the regeneration of epithelium and cartilage and the degree of rejection were assessed by counting the monocellular infiltration. Result: Investigation at day 7, showed no difference in epithelial regeneration, however, at days 14 and 30, Group 1 showed better regeneration of epithelium than groups 2 and 3. There was no difference of epithelial regeneration between group 2 and 3. There was little rejection at day 7, but at days 14 and 30, there was significant rejection in group 2 and group 3.(P<0.05). Group 3 showed lesser rejection than group 2 at days 14 and 30, but it was not statistically significant. Cartilage showed no rejection and maintained its viability in groups 2 and 3. Conclusion: Cryopreserved tracheal homograft can maintain its viability, therefore it may represent a possibility of clinical application for tracheal replacement. However, cryopreservation can not eliminate the antigenicity of the trachea completely. Furthere studies for lowering the antigenicity and rejection should be performed for an ideal substitute for tracheal replacement.
Kim, Seung-Hun;Choi, Kwang-Hwan;Lee, Dong-Kyung;Oh, Jong-Nam;Hwang, Jae Yeon;Park, Chi-Hun;Lee, Chang-Kyu
Asian-Australasian Journal of Animal Sciences
/
v.29
no.8
/
pp.1095-1101
/
2016
Ginsenoside Rg1 is a natural compound with various efficacies and functions. It has beneficial effects on aging, diabetes, and immunity, as well as antioxidant and proliferative functions. However, its effect on porcine embryo development remains unknown. We investigated the effect of ginsenoside Rg1 on the in vitro development of preimplantation porcine embryos after parthenogenetic activation in high-oxygen conditions. Ginsenoside treatment did not affect cleavage or blastocyst formation rates, but did increase the total cell number and reduced the rate of apoptosis. In addition, it had no effect on the expression of four apoptosis-related genes (Bcl-2 homologous antagonist/killer, B-cell lymphoma-extra large, Caspase 3, and tumor protein p53) or two metabolism-related genes (mechanistic target of rapamycin, carnitine palmitoyltransferase 1B), but increased the expression of Glucose transporter 1 (GLUT1), indicating that it may increase glucose uptake. In summary, treatment with the appropriate concentration of ginsenoside Rg1 ($20{\mu}g/mL$) can increase glucose uptake, thereby improving the quality of embryos grown in high-oxygen conditions.
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