Seven Years of Experiences of Preclinical Experiments of Xeno-Heart Transplantation of Pig to Non-Human Primate (Cynomolgus Monkey).

BACKGROUND: The absolute shortage of donors compared with patients requiring transplantation is currently an unsolved problem, and the only possible solution may be xenotransplantation. To establish a successful clinical trial, a preclinical study using nonhuman primates is essential. Starting in November 2011, our team initiated heterotopic abdominal heart xenotransplantation, the first in the Republic of Korea. We present here the initial 7-year results. METHODS: A total of 22 xenotransplantation procedures have been performed since 2011. Single transgenic pig (alpha-galactosidase transferase knockout [GalT KO], n = 16), double transgenic pig (GalT KO + CD46, n = 3, and GalT KO + CD39, n = 2), and triple transgenic pig (GalT KO + CD46 + CD70, n = 1) models were used. Our baseline regimen of immunosuppressants comprised CD154 ab, rituximab, anti-thymocyte globulin, tacrolimus, mycophenolate mofetil, and steroids. RESULTS: The mean graft survival was 16 ± 16.27 days, and the mean graft survival was significantly longer in cases performed since 2014 (7.5 ± 8.03 days vs 24.67 ± 17.50; P = .01). Although the donor heart ischemic time was decreased per annum, no correlations could be found between ischemic time and survival days of the graft. Double or triple genetic manipulated hearts exhibited significantly better survival (11.63 ± 11.29 days vs 30.83 ± 20.34 days; P = .03). When the ratio of heart weight (grams) to nonhuman primate weight (kilograms) was lower, the results tended to be better (P < .05). The rate of immediate postoperative bleeding (9%, n = 2) causing death was relatively high in the earlier period, but there have been no serious surgical complications affecting graft survival since 2013. CONCLUSIONS: Investigation of effective and optimal target genes for each organ to further progression toward better results is important. In addition, the immunosuppressive regimen needs to be further studied and constantly refined.

Outcomes of alpha 1,3-GT-knockout porcine heart transplants into a preclinical nonhuman primate model.

BACKGROUND: Solid organ xenotransplantation is a potential solution to current organ shortages in allotransplantation. We performed four heart transplantations from alpha1, 3-galactosyltransferase gene-knockout (GT-KO) pigs to cynomolgus monkeys and monitored immunological parameters before and after transplantation. METHODS: After blood typing of the cynomolgus monkeys, we assessed the binding activity of immunoglobulin G (IgG) and IgM of monkey serum and serum toxicity toward porcine peripheral blood mononuclear cells (PBMCs) using flow cytometry. Immunosuppressive protocols consisted of anti-thymocyte globulin (25 mg/kg), rituximab (20 mg/kg), anti-CD154mAb (20 mg/kg), cobra venom factor (0.05 mg/kg), tacrolimus, and steroid. Cynomolgus monkeys with A or AB blood type with the lowest antibody binding and serum toxicity activity on porcine PBMCs were selected as recipients. RESULTS: Absolute numbers of CD3(+) T cells, CD20(+) B cells, and CD3(+)CD95(+) memory T cells in the peripheral blood were suppressed upto 24 days after transplantation. Interferon gamma production of T cells in response to porcine antigens were also significantly suppressed. Heart xenografts from GT-KO pigs survived for upto 24 days without pathologic evidence of rejection. CONCLUSION: We successfully performed 4 heart xenotransplantations using GT-KO pigs. We overcame hyperacute rejection by using GT-KO pigs, and all of the heart xenografts from the GT-KO pigs survived between 11 and 24 days without pathologic evidence of rejection, disseminated intravascular coagulation, or consumptive coagulopathy; however, we need to optimize protocols for immune modulation and postoperative care to attain long-term survival of solid organ xenografts.

Ultrasonographic evaluation of costal cartilage fractures unnoticed by the conventional radiographic study and multidetector computed tomography.

INTRODUCTION: Rib fractures are the most common injuries resulting from blunt chest trauma. However, costal cartilage fractures are almost invisible on chest X-rays unless they involve calcified cartilage. The sensitivity of conventional radiography and computed tomography for detecting rib fractures is limited, especially in cases where rib cartilage is involved. Therefore, this study was designed to evaluate the sensitivities of chest wall ultrasonography, clinical findings, and radiography in the detection of costal cartilage fractures. MATERIALS AND METHODS: A total of 93 patients presenting with a high clinical suspicion of rib or sternal fractures were recruited for radiological workup with posterior-anterior (PA) chest radiographs, oblique rib views, sternal views, computed tomography, and chest ultrasound between April 2008 and May 2010. There were 47 men and 46 women, and the mean age of the patients was 51.8 ± 15.9 years (range 17-78 years). These patients with minor blunt chest trauma showed no evidence of rib fractures on conventional radiography and computed tomography, and no evidence of other major fractures. Chondral rib fractures were detected by using ultrasonography on a 7.5-MHz linear transducer. RESULTS: Of the total 93 patients, 64 (68.8%) showed chondral rib fractures, whereas 29 (31.2%) did not. The mean number of chondral rib fracture sites detected in 64 patients was 1.8 ± 0.8 (range 1-5). Subperiosteal hematoma was the most common finding associated with costal cartilage fractures (n = 14, 15.0%), followed by sternal fracture (n = 9, 9.7%). However, subperiosteal hematoma was also noticed in 1 (1.1%) of the patients without costal cartilage fractures, and sternal fractures in 7 patients (7.5%). DISCUSSION: The results of this study suggest that ultrasonography may be a useful imaging method for detecting costal cartilage fractures overlooked on conventional radiographs and computed tomography in patients with minor blunt chest trauma. Early ultrasonographic evaluation can give more accurate information than clinical and radiologic evaluation in detecting costal cartilage fractures and sternal fractures that are overlooked on conventional radiography and computed tomography after minor blunt chest trauma.

Does near-infrared spectroscopy provide an early warning of low haematocrit following the initiation of hypothermic cardiopulmonary bypass in cardiac surgery?

This study investigated 151 patients undergoing cardiac surgery to determine whether measurement of regional cerebral oxygen saturation (rS(c)O(2)) using near-infrared spectroscopy (NIRS) can indicate a low haematocrit after initiation of hypothermic cardiopulmonary bypass (CPB). Haematocrit, rS(c)O(2), haemoglobin level, arterial partial pressures of carbon dioxide and oxygen, systemic blood pressure, and nasopharyngeal and rectal temperatures were determined 5 min after the initial administration of heparin for CPB and 90 s after completion of the first cardioplegic solution injection. Immediately after initiation of hypothermic CPB, rS(c)O(2), haemoglobin and haematocrit values were significantly lower than those before CPB. No significant correlations were found between the change in haematocrit and changes in left, right and mean rS(c)O(2); thus, changes in rS(c)O(2) before and after initiation of hypothermic CPB did not reflect changes in haematocrit values. This indicates that NIRS cannot provide early warning of a low haematocrit immediately after initiation of hypothermic CPB in cardiac surgery.

Myocardial injury in remifentanil-based anaesthesia for off-pump coronary artery bypass surgery: an equipotent dose of sevoflurane versus propofol.

This randomised controlled trial compared the effect of equipotent anaesthetic doses of sevoflurane (S group) versus propofol (P group), during remifentanil-based anaesthesia for off-pump coronary artery bypass surgery, on myocardial injury. Either sevoflurane or propofol was titrated to maintain bispectral index values between 40 and 50. In both groups, a targeted concentration of remifentanil 20 ng x ml(-1) was maintained during anaesthesia. The concentrations of creatine kinase MB and troponin I were measured before the start of surgery, on admission to the intensive care unit, and at 12 and 24 hours after intensive care unit admission. The postoperative values of creatine kinase MB (S group: 15.08 +/- 18.97, 20.78 +/- 20.92, 12.76 +/- 12.82 vs 2.09 +/- 1.54 ng x ml(-1); P group: 10.99 +/- 13.15 27.16 +/- 56.55 11.88 +/- 18.80 vs 1.84 +/- 1.67 ng x ml(-1)) and troponin I (S group: 3.56 +/- 5.19, 566 +/- 7.89, 3.35 +/- 4.55 vs 0.52 +/- 1.90 ng x ml(-1); P group: 2.42 +/- 3.33, 4.11 +/- 6.01, 3.04 +/- 5.31 vs 0.43 +/- 1.28 ng x ml(-1)) were significantly higher than preoperative values in both groups but there were no significant differences between the two groups. There were no significant differences in time to extubation (S group, 476 +/- 284 minutes; P group, 450 +/- 268 minutes) and intensive care unit length of stay (S group, 2775 +/- 1449 minutes; P group, 2797 +/- 1534 minutes) between the two groups. In conclusion, sevoflurane and propofol at equipotent doses guided by bispectral index with remifentanil 20 ng x ml(-1) had similar creatine kinase MB and troponin I values.

Endovenous laser ablation of the incompetent small saphenous vein with a 980-nm diode laser: our experience with 3 years follow-up.

PURPOSE: To demonstrate the long-term treatment outcomes of endovenous laser ablation (EVA) of incompetent small saphenous veins (SSV) with a 980-nm diode laser. MATERIALS AND METHODS: Eighty-four patients (96 limbs), with varicose veins and reflux in the SSV on duplex ultrasound examination, were treated with a 980-nm diode laser under ultrasound- or fluoroscopy-guidance. Patients were evaluated at 1 week and 1, 3, 6 months, 1 year and yearly thereafter. RESULTS: In the 96 limbs, the technical success rate was 100%. The SSV remained closed in 89 of 93 limbs (96%) after 1 month, all of 82 limbs after 6 months, 77 limbs after 1 year, 71 limbs after 2 years and 55 limbs after 3 years. In four limbs where recanalisation was observed, repeat EVA was done resulting in successful obliteration of the SSV. No major complication occurred however bruising (27%), tightness or pain (13%) and paraesthesia (4.2%) were observed. CONCLUSION: Endovenous laser ablation with a 980-nm laser wavelength is an easy and safe procedure in incompetent SSVs. After successful treatment, there is a very low rate of recanalisation of the SSV, which suggests that the procedure will provide lasting results.

Thrombogenicity of mechanical aortic valves in an animal model: site specific testing is crucial.

We evaluated a new trileaflet prosthesis and a control bileaflet prosthesis in the mitral and aortic positions in 27 calves. The prototype trileaflet valve (TV1) functioned satisfactorily in the mitral position (TV1m, n = 7) but later yielded thrombogenic complications in the aortic position (TV1a, n = 4). The valve was redesigned (TV2) and retested in the mitral (TV2m n = 4) and aortic (TV2a, n = 5) positions, along with control valves (Cm, n = 4; Ca, n = 3). At necropsy, the valves were graded on a scale of 0 (no visible thrombi) to 4 (thrombi greater than 5 mm and/or obstructed leaflets). The TV1m, TV2m, and Cm animals, respectively, had implant durations of 215+/-112, 140+/-63, and 159+/-89 days and thrombus grades of 0.71+/-0.76, 0.33+/-0.58, and 1.50+/-0.58. The TV1a, TV2a, and Ca animals had implant durations of 18+/-12, 159+/-61, and 108+/-62 days and thrombus grades of 2.75+/-1.00, 0.50+/-0.58, and 0.67+/-0.58 (p < .005; TV2a vs. TV1a). Three TV1a calves died early of valve related complications. A design irregularity, undetected in the mitral position but revealed in the aortic position, caused a high early mortality in the TV1a animals. Redesigning the prosthesis eliminated valve related mortality and significantly reduced the thrombus grade. Because satisfactory performance in the mitral position does not guarantee safety or efficacy in the aortic position, site specific preclinical testing is crucial for mechanical heart valves.

Rabbit model of cardiopulmonary bypass.

Mainly because of technical problems, the use of rabbits as a cardiopulmonary bypass (CPB) animal model with direct cannulation of the ascending aorta is known to be extremely difficult. The objectives of this study were the establishment of a CPB model in rabbits with direct cannulation of the ascending aorta, and the evaluation of the protective effect of steroid on the development of brain edema during circulatory arrest (CA) in an established rabbit CPB model. Fifteen New Zealand white rabbits were divided into three groups; control CA group, CA with Trendelenberg position, and CA with Trendelenberg position and steroid administration. After anesthetic induction and tracheostomy, median sternotomy was performed. An aortic cannula (3.3 mm) and a venous cannula (14 Fr) were inserted into the ascending aorta and the right atrium, respectively. The CPB circuit consisted of a roller pump and a bubble oxygenator. With 120-150 ml of blood, the priming volume of the circuit was approximately 450 ml, and CPB at a flow rate of 80-85 ml/kg/min was initiated. Blood in the priming solution was obtained from donor rabbits through cardiac puncture. Ten minutes later, CA with cessation of CPB was established for 40 min at 20 degrees C (rectal temperature). After CA, CPB was restarted with a 20 min period of rewarming. Ten minutes after weaning, the animal was sacrificed. Between 1 and 2 g of the brain was removed and the water content was determined and compared between groups. CPB with CA was successfully performed in all cases, with a flow rate of 60-100 ml/kg/min maintained throughout the CPB procedure. At that time, blood gases were reasonably maintained and aortic pressure ranged from 35 to 55 mmHg. After weaning from CPB, all hearts resumed beating spontaneously. Among the three groups, there were no statistically significant differences in the water content of the brain. These results indicate that: (1) if the proper technique is used, CPB in rabbits with direct cannulation of the ascending aorta is a reliable procedure, and (2) the effect of steroid on the prevention of brain edema related to the Trendelenburg position during CA is not established within the scope of this study.