Heterotopic Transplantation Model Research Articles

Anti-allergic agent tranilast decreases development of obliterative airway disease in rat model of heterotopic tracheal transplantation

Background Tranilast is an anti-allergic agent known to inhibit the release of histamine, interleukin-1β, transforming growth factor β1, and platelet-derived growth factor from various cells and currently is used to treat allergic diseases, keloids, and hypertrophic scars. We evaluated the ability of tranilast to inhibit the development of obliterative airway disease (OAD) in a rat model of heterotopic tracheal transplantation. Methods We transplanted tracheal segments from donor rats (Brown Norway) into subcutaneous pouches in major histocompatibility complex-incompatible recipient rats (Lewis). At Days 21 and 28 after transplantation, we histologically assessed the harvested allografts scored the degree of OAD, on a scale from zero to 4 as previously described, caused by fibroproliferative tissue. Results Recipient animals treated orally with 400 mg/kg/day tranilast throughout the experiment showed significantly decreased OAD compared with control animals, with a histologic score of 1.1 ± 0.4 vs 3.0 ± 1.3, respectively (mean ± SD, p = 0.007), at Day 21 after transplantation and 2.0 ± 1.4 vs 3.9 ± 0.4, respectively (mean ± SD, p = 0.017), at Day 28 after transplantation. Conclusion These results showed that treatment with tranilast significantly decreased fibroproliferative airway changes associated with allograft rejection in a rat model of tracheal transplantation, suggesting that tranilast may be useful in preventing bronchiolitis obliterans after lung transplantation.

The effects of pretreatment with donor antigen and immunosuppressive agents on fully allogenic tracheal graft

Obliterative bronchiolitis is a major clinical problem in cases involving a transplanted lung. We examined drug-induced tolerance to a fully allogenic tracheal graft in a murine heterotopic transplantation model. Recipient mice (C57BL/6) were primed iv with 1 x 10(8) splenocytes of donor mice (BALB/c). Day 0 was the day of the splenocyte injection. Cyclophosphamide and Busulfan were injected intraperitoneally on day 2. On day 3, 1 x 10(7) donor bone marrow cells were intravenously injected. On day 28, a donor tracheal graft was implanted into a subcutaneous pocket. Grafts were harvested at 3-week intervals, and the degree of obstruction of the inner cavity, the condition of epithelium, and the viability of chondrocytes were examined. All of the isograft controls (BALB/c) and grafts implanted in the T cell-free recipients (BALB/c-nu) showed patent, lined epithelium and viable chondrocytes. All allografts tested showed total luminal occlusion by granulative tissue and inflammatory cells, and the epithelium was totally absent. Five of 11 drug-treated grafts were completely patent, although the epithelium was almost absent and the chondrocytes were substantially destroyed. However, when the chimerism was analyzed by flow cytometry analysis of the recipient T cells, approximately 90% of the donor cells were recognized. Even by this pre-treatment-induced chimerism, a transplanted allogenic trachea was not completely preserved. The present results suggest that a non-allogenic response might have contributed to the rejection.

A New Experimental Model of Heterotopic Intrathoracic Heart Transplantation without Cardiopulmonary Bypass

Background and Objectives:A variety of experimental heterotopic heart transplantation models have been developed for the purpose of this study. However most were complicated and used extracorporeal circulation. A new and simple experimental working heterotopic intrathoracic heart transplantation model has been developed in dogs, which could be performed without support of extracorporeal circulation. Materials and Methods:Six mongrel dogs were used for the three heterotopic heart transplantation experiments. The heterotopic hearts were transplanted in the right thoracic cavity using a right thoracotomy. The superior vena cava and left atrial cuff of the donor heart was anastomosed to the superior vena cava and left atrium of the recipient heart. Consequently, the aorta and pulmonary artery were anastomosed to the ascending aorta and right atrial appendage of the recipient heart, without cardiopulmonary bypass. The electrocardiograms of the donor and recipient hearts were measured by electrodes placed at the ventricular apexes and on the skin. Results:The donor hearts survived for 10, 6 and 18 days after transplantation, without any immunosuppressive agents or anticoagulants. The electrocardiograms of both hearts could be measured using the electrodes. There were no wave or voltage changes on electrocardiography. Conclusion:This working heart model of heterotopic intrathoracic transplantation, without cardiopulmonary bypass, may be useful in studies for various purposes, i.e., the effects of different pharmacological agents on the conduction system of the denervated heart and the hemodynamic changes of the recipient with auxiliary support of the transplanted heart. (Korean Circulation J 2004;34(6):593-599)

Heterotopic limb allotransplantation model to study skin rejection in the rat

Current rodent models for investigation of limb allotransplantation typically utilize orthotopic whole-limb transplantation, a morbid and time-consuming procedure. Our objective was to design a less morbid rat model to explore the immunological obstacles of limb transplantation, and particularly skin. Twenty lower hindlimbs from 10 donors were transplanted into a heterotopic subcutaneous position into 20 animals (10 isogeneic and 10 allogeneic). Each group was further subdivided to include animals with (n = 5) and without (n = 5) a skin paddle for observation of cutaneous signs of rejection. All grafts in the isogeneic group survived for 100 days, i.e., the endpoint of the study. Allogeneic transplants rejected their allografts at a mean of 12.8 days (with skin) and 20.6 days (without). Our heterotopic limb transplantation model takes less time and is less stressful to the animals, while allowing for early observation of graft skin rejection, when compared to orthotopic whole-limb transplantation.

Reduction of recipient macrophages by gadolinium chloride prevents development of obliterative airway disease in a rat model of heterotopic tracheal transplantation.

Recent studies have shown the possible role of growth factors and the involvement of macrophages as a source of them in the pathogenesis of bronchiolitis obliterans (BO) after lung transplantation. The authors intended to determine whether depletion of recipient macrophages by gadolinium chloride (GdCl3) resulted in decreased obliterative airway disease (OAD) in a rat model of heterotopic tracheal transplantation. A tracheal segment of donor rats (Brown Norway) was transplanted into a subcutaneous pouch of fully major histocompatibility complex-incompatible recipient rats (Lewis). Recipients were injected intravenously with 80 mg/kg of GdCl3.6H2O or saline on days 0, 7, and 14 posttransplant. Allografts were harvested on days 7, 14, 17, and 21 and the degree of OAD resulting from fibroproliferative tissue was pathologically scored on a scale of 0 to 4. A portion of allografts was submitted to reverse-transcriptase polymerase chain reaction analysis to examine mRNA expression for platelet-derived growth factor (PDGF), basic fibroblast growth factor, and transforming growth factor-beta1. Immunohistochemical studies confirmed reduction in the number of ED2+ macrophages in tracheal allografts by GdCl3 injection. GdCl3 treatment significantly decreased OAD of allografts, with the histologic score of 1.4+/-0.3 in the treated animals compared with 3.0+/-0.5 in the controls (mean+/-SE, P=0.02) at day 21 posttransplant, and this was accompanied by decreased PDGF-A and PDGF-B gene expression in the GdCl3 group at day 17 posttransplant. Macrophage reduction by GdCl3 resulted in significantly decreased OAD development and reduced PDGF mRNA expression in allografts. This suggests a potential effectiveness of therapies targeting recipient macrophages in preventing BO after lung transplantation.

Description of a new auxiliary heterotopic partial liver transplantation technique with portal vein arteriolization of applicability in heterotopic liver xenotransplantation

Objective The aim of this study was to describe a new model of auxiliary heterotopic partial liver transplantation with portal vein arterialization. Materials and methods Three standard hepatectomies were performed in pigs. The left lateral lobe was surgically resected and portal vein arteriolization constructed by an end-to-side “Y” anastomoses between the distal to the celiac axis aorta and the portal vein. Results The graft was placed in the left iliaca fossa using anastomoses of the donor infrahepatic inferior cava vein end-to-side to the host infrarenal inferior vein and the donor aortic stump with portal vein arteriolization end-to-side to the left iliac artery. After graft reperfusion, the 3 recipients showed intraoperative hypotension, which was treated with fluid administration and vasoactive drugs. At the end of the operation, the graft displayed normal arterial blood flow and good venous drainage. The donor liver graft appeared more red than the host liver, which was due to the increased arterial blood flow. One pig of 3 died at 24 hours after surgery, probably due to hypothermia. However, the other 2 pigs survived the procedure and remained stable. Echographic monitoring showed intrahepatic arterial expansion, which may be the result of high blood pressure due to the arteriolization procedure. Conclusions We have developed a novel and easy to perform technique that diminishes the number of anastomoses and does not involve vessels from other organs.

Mouse model of heterotopic aortic arch transplantation

Background. Syngeneic heterotopic transplantation of segments of descending thoracic aortas containing atherosclerotic lesions from hypercholesterolemic mice into normocholesterolemic recipients has been useful for studies on plaque regression and stabilization. Because lesion development is more rapid and exuberant in the aortic arch, a technique of transplantation of the mouse aortic arch was developed. Materials and methods. C57BL/6, apoE-deficient (apoE-/-) (hypercholesterolemic) mice were fed a Western diet for 22 weeks and used as donors of aortic-arch segments containing atherosclerotic lesions. Twenty syngeneic transplants were performed on age-matched wild-type (normocholesterolemic) mice. Aortic arches containing atherosclerotic lesions were implanted on the abdominal aorta of recipient mice by end-to-side microsurgical anastomosis. Two weeks after transplantation, grafts were noninvasively imaged in vivo by magnetic resonance (MR) microscopy. Grafts harvested four weeks after transplantation were submitted for histological examination. Results. All recipients survived the entire follow-up period (1 month) without complications. Duration of recipient procedure ranged from 90 to 120 (mean, 105) min; aortic clamping time varied from 45 to 60 min. In vivo MR microscopy demonstrated patency of the grafts and wall thickening that corresponded to the preexisting atherosclerotic lesions. Histology confirmed patency and atherosclerotic thickening of the grafts, and showed no evidence of acute tissue damage. Conclusions. Syngeneic transplantation of the aortic arch in mice represents a useful alternative model for studies on morphology, imaging, and mechanisms of atherosclerosis. The curvature of the aortic arch is preserved after implantation onto the abdominal aorta, providing clear landmarks for noninvasive assessment using MR.

An experimental study on the growth of condylar cartilage, using a new vascularized mandible heterotopic transplant model

Purpose: To clarify the influence of external and dynamic factors on the growth of mandibular condyle, we developed vascularized mandible heterotopic transplant (VMHT) model. In this report, we discuss histologic changes in the mandibular condylar cartilage in the absence of external factors, using VMHT models. Materials and Methods: Thirty VMHT model using male Lewis rats aged 6 weeks were produced. Six VMHT rats were randomly selected for death at 1, 2, 4, 7, and 14 days after transplantation. We histologically investigated the mandibular condyles of the grafts and controls, the left mandibles of recipient rats. Results: In the transplanted mandibular condyle of VMHT models, the cell arrangement became increasingly irregular with the passage of time, with a concomitant, irregular thickening or thinning of the cartilage layer. There was heterotopic fibrous ossification, extending horizontally under the proliferative cell layer in these regions of thin cartilage. After these events, cartilage cells had disappeared almost completely by 14 days after transplantation. Conclusions: This study indicated that external and dynamic factors are not necessary for the differentiation and proliferation of the condylar cartilage cells. Instead, these factors affect the maintenance of the orderly growth of the cartilage cells and may serve a critical role in the differentiation of undifferentiated mesenchymal cells into chrondroblasts. © 2003 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 61:239-245, 2003

A new vascularized mandible heterotopic transplant model for studies on the growth of condylar cartilage

Purpose: We developed a vascularized mandibular heterotopic transplant (VMHT) model to allow the observation of condylar cartilage growth in the absence of external and dynamic factors, such as mastication. Materials and Methods: In VMHT, we performed a transplantation between 2 age-matched inbred rats (male Lewis rats aged 6 weeks). The hemimandibular tissue, with the common carotid artery and external jugular vein as vascular pedicle, was harvested from the donor rat and transplanted to the neck of the recipient rat with the use of vascular anastomosis under an operating microscope. To evaluate the stability of VMHT, we investigated both the weight change of the recipient rats and the blood circulatory conditions of the grafts via the radioactive microsphere method, laser Doppler blood flowmeter, and angiography. Results: Although the body weight of the recipient rats decreased immediately after transplantation, these values subsequently increased, indicating a good nutritional environment in the recipient rat. Also, the VMHT graft tissue and the mandibular condyle had favorable blood circulation, equivalent to normal conditions. Conclusions: In VMHT, transplantation between the 2 age-matched inbred rats using anastomosis of the similar blood vessels enables us to minimize the influences of genetic factors, hemodynamics, hormones, and other external factors. Our investigation indicated that the transplant possessed favorable growth conditions, equivalent to the environment of the endogenous mandibular condyle. Thus, we are able to maintain conditions similar to the normal environment of the mandibular condyle. This model will be very useful in future investigations of the influences of external and functional factors on chondrogenesis and enchondral ossification of the mandibular condyle. © 2003 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 61:234-238, 2003

Heterotopic transplantation of cryopreserved tracheae in a rat model.

The successful use of cryopreserved tracheal allografts in canine models suggests their use in humans. The grade of genetic difference, the mechanism of revascularisation and the method of cryopreservation are not clearly defined. The purpose of our study was to investigate the rejection of tracheal transplants in a standardised heterotopic rat model using different forms of cryopreservation. Tracheae from Brown Norway rats were implanted into the omentum from Brown Norway rats or Lewis rats. We transplanted fresh isografts or allografts and pretreated isografts or allografts. Cryopreservation was performed in a medium containing 10% dimethyl sulphoxide at -80 degrees C for 28 days (I) or -196 degrees C for 84 days (II) or without medium at -80 degrees C for 28 days (III). The transplants were excised after 7 and 21 days, respectively. Histological examinations revealed normal structure and function of isografts after 21 days. In the cryopreserved isograft, the epithelium had disappeared and the tracheal lumen was partially obstructed by a non-compact fibrous tissue. In the fresh allografts, the epithelium was replaced by aggressive fibrous tissue, infiltrating the membranous part of the trachea and occluding the tracheal lumen. The cartilage was vital without any sign of rejection. In the cryopreserved allografts, the tracheal lumen was obstructed by dense fibrous tissue with an inflammatory reaction. The cartilage of cryopreserved allografts (II) and (III) had lost the nuclei corresponding to non-vital tissue. Only in the cryopreserved allografts (I) did we find nodular regeneration at the edges of the cartilaginous bow. The heterotopic transplantation model allows the study of the mechanisms leading to tracheal obstruction. Cryopreservation was found to have no clear advantage in reducing transplant immunogenicity. Cryopreservation leads to significant damage to the cartilage, the intensity of which is dependent on the mode of cryopreservation.

Experimental model of heterotopic uterus transplantation in the laboratory rat.

The present study describes a standardized experimental model of whole-uterus-and-ovaries heterotopic allotransplantation in the laboratory rat. Fifteen transplantation procedures were done. The anatomy of the pelvic region was studied with an additional 20 dissections, noting the topographical and vascular anatomy of the uterus, fallopian tubes, and ovarian vessels. Recipients were randomized into three groups. The average operative time was 150 min. The postoperative survival rate was 100%. Postoperative vascular anastomosis patency was 100%, and 26% at 72 hr. Recipients were euthanized at 24 hr (group I), 48 hr (group II), and 72 hr (group III); grafts were harvested and examined macroscopically, and fixed in formaline for histopathological and immunocytochemical analysis. Failure in 74% of the grafts at 72 hr was due to early thrombosis, starting from the capillary bed and progressing towards the main feeding pedicles. More studies must be undertaken to further understand the rejection mechanisms in transplanted reproductive organs. The efficiency, feasibility, and safety for such an operation in humans remain to be proven. We consider the present model a suitable tool to study all the above-mentioned goals.

The effect of selective inhibition of cyclooxygenase (COX)-2 on acute cardiac allograft rejection.

Using a rat (Lewis-Wistar Furth) abdominal heterotopic transplantation model, we reported previously that the expression of cyclooxygenase (COX)-2 is increased in parallel with that of nitric oxide synthase (NOS)-2 during cardiac allograft rejection. To investigate effects of COX-2 inhibition in this model, allograft recipients were treated orally (PO) with 5 mg/kg per day of the tetra substituted furanone selective COX-2 inhibitor 5,5-dimethyl-3-(3 fluorophenyl)-4-(4 methylsulfonal) phenyl-2 (5H)-furanone (DFU) in 1% methyl cellulose solution. In the treated animals, allograft survival was increased from 6.3+/-0.5 to 12.6+/-2.6 days (P = .001). At days 3 and 5 posttransplantation, there were reductions in the extent of the inflammatory infiltrate, endovasculitis, myocardial edema, and cardiomyocyte damage in rejecting allografts. The mean numbers of apoptotic cardiomyocytes determined with the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) technique were significantly reduced in DFU-treated grafts compared with untreated controls (P < 0.05). At day 3 posttransplantation, prostaglandin E2 synthesis by myocardial slices incubated with 100 microM bradykinin was reduced from 1,097+/-156 to 153+/-63 pg/mg of protein in the treated allografts (P < .005). At day 5, COX-2 protein and mRNA together with COX-2, NOS-2, and nitrotyrosine immunostaining in damaged cardiomyocytes were diminished in treated versus control grafts. The data indicate that the inhibition of COX-2 prolongs allograft survival and reduces myocardial damage and inflammation during acute cardiac allograft rejection.

Pirfenidone: A Novel Anti-fibrotic Agent and Progressive Chronic Allograft Rejection

In our established model of heterotopic tracheal transplantation, at day 28 following transplantation, obliteration of the lumen is observed, which is histologically similar to that seen in Obliterative Bronchiolitis (OB). Pirfenidone (Pir) is a novel anti-fibrotic agent that causes no immunosuppression, but does downregulate the production of TGF-beta and collagen in vitro. We hypothesized that when used in this in vivo model, that Pir may alter the observed luminal fibrosis and obliteration. Methods: The treatment groups were: CSA, Pir and CSA, Pir only (n=6 each). Luminal supernatants and tissue were obtained from these groups at day 28. H&E staining was completed, as well as MTS proliferation assays, and TGF-beta ELISA on the fluids. Results: The CSA-Pir combined treatment group was the least fibrogenic in vitro ( p<0.001). The TGF-beta levels were elevated in all groups (range 203–372 pg/ml). The H&E staining revealed that the luminal obliteration was less organized in the combined CSA-Pir group. Conclusions: Our study shows that the combination of CSA-Pir resullts in a less fibrogenic luminal fluid and a less dense fibrous luminal plug. Pir should be further studied in obliterative airways disease (OAD).

Successful 24-h Preservation of Canine Small Bowel Using the Cavitary Two-layer (University of Wisconsin Solution/Perfluorochemical) Cold Storage Method

We previously developed the two-layer cold storage method (TLM), which allows sufficient oxygen delivery to the canine pancreas during preservation, and successfully achieved 96-h preservation. In this study, we applied a modified TLM (cavitary TLM) to small bowel preservation in a canine heterotopic transplant model. Using simple storage in University of Wisconsin solution (UWM, group 1, n = 12) or cavitary TLM (group 2, n = 8), 40 cm segments of the jejunum were preserved for 24 h at 4 degrees C. The nonpreservation group served as the control (group 3, n = 8). The grafts were implanted heterotopically as a Thiry-Vella loop. Eleven of 12 dogs in group 1 died within 3 days post-transplant as a result of graft intraluminal hemorrhage, while all dogs in groups 2 and 3 survived until day 7. Histological analyses showed almost normal structures of the graft mucosa in groups 2 and 3 at day 7. Results from maltose and acetaminophen absorption tests in group 2 were comparable to those in group 3. Only one survivor in group 1 showed distinct graft mucosal damage, confirmed by histological and functional analyses. In our transplant model, the canine small bowel was successfully preserved by cavitary TLM for at least 24 h, while this preservation time was beyond the limit with UWM.

Modified techniques of heterotopic total small intestinal transplantation in rats.

To establish a successful model of heterotopic total small intestinal transplantation (SIT) in rats in order to reduce the complications and increase the survival rate. A total of 196 Wistar rats underwent heterotopic SIT with microsurgical technique. Technical modifications included shortening fasting time and supplying energy before surgery, administering optimal volume of crystalloid fluid to the donor and recipient during surgical procedures, reducing mechanical and ischemic injuries to donor intestine, revascularizing small intestinal graft with a combination of conventional aorta to aorta anastomosis and a cuffed portal vein to left renal vein anastomosis which resulted in an acceptably short warm ischemic time, and also an adequate blood supply and drainage of the graft. The average time for the donor surgery was 86 min +/- 20 min, the mean operative time for the recipient was 115 min +/-20 min and warm ischemia time was shortened to 40 min +/- 5 min. There was a shorter revascularizing time of the graft, the abdominal aorta (AA) to AA anastomosis being 21 min +/- 10 min, and the cuffed portal vein (PV) to the renal vein anastomosis being 5 min +/- 5 min. The one-week survival rate of 98 rats with SIT was 88.78% (87/98), without thrombosis and stenosis of anastomosis. The longest survival time of recipient rats was more than 389 days after SIT, the rats were maintaining normal weight, with perfect intestinal function and intact intestinal histology. These modified techniques for SIT would remarkably reduce the complications and improve survival rate in rats, which provided a potentially more consistent and practical model for experimental and clinical studies.

Dependence of murine obstructive airway disease on CD40 ligand.

Human lung transplantation carries a poor prognosis because of chronic rejection in the form of obliterative bronchiolitis syndrome (OBS). Using the mouse model of heterotopic tracheal transplantation, we examined the role of costimulation in the allograft rejection that characterizes obstructive airway disease (OAD). C57BL/6 or BALB/c tracheae were implanted into wild-type control, CD28-/-, muMT (B-cell deficient), or CD40L-/- recipient mice. Grafts were explanted from 7 to 42 days posttransplantation and evaluated. Thickening of the basement membrane and a decrease in patent luminal area were first noted at 2 weeks in wild-type allogeneic trachea recipients and to a slightly lesser degree in CD28-/- recipients. In contrast, CD40L-/- recipient mice showed no evidence of cellular infiltrates or fibrosis in transplanted tracheae. To determine whether CD40L interacted with host or donor CD40, CD40-deficient tracheae were transplanted into CD40L+/+, CD40+/+ wild-type mice. Wild-type mice rejected CD40-/- tracheae. Tracheae were transplanted into B-cell-deficient mice to determine the role of B-cell CD40 in chronic pulmonary allograft rejection. The OAD reaction was identical in wild-type and B-cell-deficient mice. Development of OAD in the mouse trachea transplant model is primarily dependent on CD40L and is relatively CD28 independent. The ability of mice to reject CD40-/- tracheae demonstrated that host, not donor, CD40 is required for rejection. Furthermore, the ability of B-cell-deficient mice to reject allogeneic tracheae demonstrated that B-cell CD40-mediated responses are not required for the development of OAD.

T1 relaxation times for viability evaluation of the engrafted and the native liver in a rat model of heterotopic auxiliary liver transplantation: a pilot study.

Following a heterotopic auxiliary liver transplantation, commonly used measurements are either invasive or non-indicative of individual viability of the coexisting engrafted and native livers. Magnetic resonance imaging (MRI) was therefore tested for its potential to monitor the post-transplant hepatic viability in a rat model. Thirteen Wistar rats were systematically evaluated with MRI and serum biochemical liver parameters. Post-transplant complications and the causes of animal death were identified by autopsy and histo-pathological examinations. The data of the healthy survivors were compared with those of the rats that developed complications. On MRI, the hepatic complications could be depicted in the individual livers. A specific pattern of signal evolution was found in the livers of the healthy survivors: the mean T1 relaxation times of the engrafted livers increased immediately after transplantation (476 +/- 64 ms, mean +/- standard deviation, pre-operative; 730 +/- 48 ms, week 1) and then declined steadily to a 3 month value of 489 +/- 246 ms, while, following a transient first rise (476 +/- 64 ms, pre-operative; 589 +/- 28 ms, week 1), the mean T1 value of the native livers increased again 4 weeks after surgery and reached a 3 month value of 859 +/- 43 ms. However, in the rats with various complications, the mean T1 relaxation times of the engrafted livers continued to increase throughout the first post-operative month (760 +/- 48 ms, week 1; 922 +/- 76 ms, week 4), while that of the native liver only varied mildly (546 +/- 25 ms, week 1; 473 +/- 25 ms, week 4). After the first post-transplant week, the healthy engrafted livers could already be distinguished from those with complications by a significant decrease in T1 relaxation times. These data suggest that, besides demonstrating major complications, MRI may allow one to monitor the viability of each liver by analysing the relative signal intensity and T1 relaxation times after a heterotopic auxiliary liver transplantation.

Apoptosis and rejection in rat intestinal transplantation: correlation with FK506 doses and donor specific bone marrow infusions.

Our purpose was to investigate the occurrence and the evolution of apoptosis of enterocytes during acute and chronic rejection in an experimental model of allogeneic heterotopic small bowel transplantation (SBTx). Forty-five rats were divided in 10 experimental groups according to the dose of FK506 administration and donor bone marrow infusions (DBMI). Groups 1 and 2 did not received BMI. Groups 3 and 4 received 150x106 cells at day 0, groups 5 and 6 received 75x106 cells at days 0-4, groups 7 and 8 received 75x106 cells at days 4 and 10, and groups 9 and 10 received 30x106 cells at days 4, 10, 15, 20, and 25. Animals of groups 1, 3, 5, 7, and 9 were immunosuppressed with 0.5 mg/kg FK 506, although the remaining groups with 1 mg/kg FK 506, from day 0 to 4 after transplant. Fragment end labeling of DNA was used to detect apoptosis. The number of apoptotic cells detected was highest at day 15 (184+/-154) and then progressively decreased thereafter (day 30=159+/-197; day 45=80+/-167; day 60=0). The number of apoptotic enterocytes was found increased during mild (151+/-108) and moderate (281+/-161) allograft rejection, although a low apoptotic rate was observed in cases without rejection (59+/-13) and during severe (53+/-131) and chronic rejection (46+/-136). Furthermore the number of labeled cells was found inversely correlated with fibrosis (P<0.0001). There was no correlation between apoptosis and the presence or absence of DBMI; however, at day 15 rats receiving 1 mg/day of FK 506 had a significantly lower number of apoptotic cells detected (127+/-103 vs. 233+/-174; P<0.02). In this study the number of apoptotic cells correlated positively with mild and moderate rejection episodes. In case of severe and chronic rejection a low apoptotic rate was found due probably to extensive necrosis and fibrosis of the mucosa. These data suggest an important role of apoptosis in acute and chronic intestinal rejection in a rat model of intestinal transplantation. Determination of apoptosis in allografts might represent an early sign of small bowel rejection and a useful marker in defining the degree of rejection and its outcome/prognosis.

Expression of proα1(I)- and proα1(III)collagen mRNA in chronic allograft rejection in a porcine heterotopic bronchial transplantation model

Expression of proα1(I)- and proα1(III)collagen mRNA in chronic allograft rejection in a porcine heterotopic bronchial transplantation model

Systolic and diastolic properties and myocardial blood flow in the heterotopically transplanted rat heart during acute cardiac rejection.

The aim of the study was to characterize the course of systolic and diastolic function, myocardial blood flow, and histologic changes during acute rejection in a model of heterotopic transplantation in rats. For this purpose isogenic Lewis-to-Lewis and allogenic DA (Dark Agouti)-to-Lewis rat cardiac transplants were studied 1 hour and 1, 3, and 5 days, respectively, after heterotopic intraabdominal transplantation. Myocardial tissue blood flow (MBF) was assessed by the hydrogen-clearance method. An implanted balloon was used to measure pressure-volume relations in the transplanted heart. Myocardial water content was determined at the end of the experiments, and histologic examinations were performed. The MBF recovered during the first day postoperatively in both groups and decreased again in the allogenic group after 3 and 5 days (p < 0.05); it remained stable in the isogenic group. Myocardial relaxation was already prolonged in the allogenic group after 3 days and deteriorated further. Left ventricular end-diastolic pressure progressively increased in the allogenic group, whereas it remained unchanged in the isogenic group up to 5 days. After recovery from ischemia, the left ventricular peak systolic pressure was stable in the isogenic group for the entire further observation period, but it significantly decreased in the allogenic group after 5 days (p < 0.05). Myocardial water content showed a significant increase in the allogenic group compared to that in the isogenic group after 5 days. In the allogenic group histologic examination confirmed mild to moderate rejection after 3 days and severe acute rejection after 5 days. Thus, after recovery from ischemia, mild to moderate cardiac rejection was associated with reduced MBF and impaired relaxation. In a typical sequence, generation of edema and impaired diastolic compliance were terminally followed by systolic dysfunction during severe rejection.