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BIOMAT., ART. CELLS & IMMOB. BIOTECH., 20(2-4), 557-561 (1992)
EVALUATION OF A PYRIDOXYLATED HEMOGLOBIN POLYOXYETHYLENE SOLUTION AS A PERFUSATE FOR SMALL INTESTINE PRESERVATION H.I.iit,
CONJUGATE
T.Agishi, T,ha*,ai,T.Hayashi, S.Fu.iita, S.Fuchinoue,li.Takahashi,
s . ' l e r a o k r i 6 h.Ota.
IJcpt .of surgery 1 1 1 , T o k y o k'omen's Yedical ('01 lege.8-1 Kawada-cho, ~ ~ r k u - k uT, o k y o 162, Japan.
Shin
ABSTTtACT new type of artificial blood, pyridoxylated hemoglobinpolgovyethglene conjuqatr( FHP) solution,(developed by PHP research group of the department of health and welfare of' Japan,and produced by Ajinomoto ro.,Inc.Tokyol a s an oxygen-carrying component, has been recently dr\,ised using hemoglobin obtained from hemolyzed human erythrocytes. Recently, the studics using this solution as a preservation sol~ttiortwere> performed in some instances. To r\amine the mechanism of improved ~ i a b i l i t y using this solution u s n preser\'ation solution, we developed a model of orthotopic s m a l l intestine transplantation ( O I T J in the rat. As a baseline study, he comparcd parameters of \lability of the grafts preserved in Collins and L'K soIutioi1 to those preser\ed i i i PHP solution including a survival rate, a st-rum level t o t a l protein and albumin, and a change in body weight after transplantation. T n o u r stiidy, the simplr hypot hermia storage together with intestinal Iwrfirsion preserlation with PHP solution was performed. ,\riimals w e r e divided into 6 , 1 3 , and 2 1 hr preservation groups. 4 1 1 of the r a t s survived after G hr preser\'ation following transplantation. However, in 1 2 hr storage, five of S I Y rats in PHP solution preser\ a t i o n sitr\.i\.eri and r e c o \ e r ) i i t hod? weight after grafting was better that, those kith Collins and L'K solution. k c C O I I C Iude that thr PHF solutiori is,therefore, considered to possit 2 1 > be H more suitable perf1csat.e for small intestine preservation than Collins and I'k solution. KEY WORDS: sinall intestine, transplantation, preservation, perfusion, pyridovglared hemoqlobin polyoxyethglene conjugate solution. ABBREVIATIONS PHP solution:Pyrido\ylnted Hemoglobin Polyoxyethylene solution OIT : 0 1 - t hot op i c in t e s t i ne t ransp 1an tat ion. IVC: lnfrrior vena cave. PV: Portal vein. UW: I'niversity of wisconsin. 4
INTRODUCTION Research and development of artificial blood as a total blood substitute has been carried out by many investigators. PHP conjugate solution as a perfusion solution for organ preservation was reported in 1986 and itsed for l i x - e r preservation( 1-3). The successful use of Cgclosporine .4 in transplantation of other vascularized organ grafts has encouraged ils to use i n small intestine transplantation.(l). However, the preservation time that allows the harvested intestine to remain viable is limited in short time by currently available techniques. If a longer preservation period is allowed for adequate transplantation, patient death due to primary graft failure is supposed to be pre\-ented. 557
Copyright 0 1992 by Marcel Dekker, Inc.
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
5 58
LIU ET AL.
At present, a tested and proven technique for small intestine preservation which is suitable for clinical application is lacking. There is an abundance of conflicting literature on experimental approaches and models of the many techniques for preserving the small intestine ranging from simple hypothermia storage in a hyperbaric oxygen chamber to pulsatile normothermic perfusion.(5-6). Recently PHP conjugate solution (stabilized hemoglobin), which is devised from hemolyzed human erythrocytes,has been developed as an artificial blood with oxygen-carrying property. It was our objective to preserve small intestine by perfusion of the intestinal lumen with 4OC PHP solution. In this study, four groups were divided by intact control(n=6), and Collins solution (n=18), UW solution(n=18), and PHP solution(n=18). The simple storage was performed with Collins and UW solution, and the additional intraluminal perfusion preservation with oxygen supply was performed with PHP solution. In each group, 6, 12, and 24 hr preservation (n=6) was carried out. In the surviving rats, serum level of total protein and albumin, a change of body weight, as parameters for viability of grafts, was checked after transplantation. MATERIALS AND METHODS
Animals, young Lewis strain rats weighting between 250 and 300g, were used both for donor and recipient in order to exclude of the immunologic problem. All of the rats were fed at Tokyo Women's Medical College Animal Experiment Center in temperature-controlled clean cages and received standard laboratory chow and rater ad libitum. Operative technique The rats were fasted for 12 hours before the operation but were allowed free access to water. They were anesthetized with ether. Donor operation: The surgical techniques were modified slightly from those originally described by Monchik and Wolfgang(7-8). The abdomen was opened a midline incision. A microscope was used to mobilize the graft. Manipulation of the intestine was tried to be minimal. An about 20cm segment of donor small intestine,the proximal jejunum apart 5cm from the ligament of Treitz, was isolated on a vascular pedicle consisting of the superior mesenteric artery with a segment of the aorta and the portal vein. The lumbar arteries from the aorta were meticulously ligated with 6-0 silk suture to minimize bleeding. After systemic heparization (15Ou/lOOg), the aorta was cannulated, and the graft vascular bed was perfused with 5-101111of heparinized lactated Ringer's solution. A small volume of cold saline solution containing gentamycin was used to irrigate the gut lumen avoiding overdistention of the donor bowel. The portal vein and the aorta above and below the superior mesenteric artery was transected with an aorta cuff, then the graft was removed. The harvested intestine was then preserved in three groups: Group 1: Control group (11.61 consisting of age and weightmatched Lewis strain rats in which no surgical procedures were performed. 2 : Collins solution: The graft vascular bed and intestinal were Qroup perfused with Collins solution and preserved for 6 hrs(n=6), 12 hrs(n=6), 24 hrs(n=6) at 4%. 3 : tJW solution: The graft vascular bed and intestinal were Qroup perfused with UW solution and preserved for 6 hrs(n=6),12 hrs(n.61, 24 hrs(n.6) at 4OC. 4: PHP solution: The graft vascular bed was Group perfused with PHP solution. Subsequently, a short wide rubber tube was inserted into the bowel for continuous perfusion preservation ( speed: 2Oml/hr )with oxygen for 6 hrs, l2hrs, and 24 hrs at 4%. Recipient operation: Recipient animals were generally anesthetized similarly to donor. The segment grafts were placed orthotopically. Revascularization was performed between the donor and recipient. The aorta and inferior vena cave were exposed, by means of a modified Lee's
559
PYRIWXYLATED HEMOGLOBIN POLYOXYETHYLENE CONJUGATE SOLUTION
I SMALL INTESTINAL PRESERVATION I (0
I1 6
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
C
H5 5 4
w
A 3
.g
x
2
czl 0
PHP
Control
Collins
(Fig. 1 )
POSTOPERATIVE BODY WEIGHT CHANGE FOLLOWING 6 Hr PRESERVATION I 0
0
1
2
3
4
WEEKS
(Fig. 2 )
SERUM TOTAL PROTEIN AND ALBUMIN CONCENTRATION 4 WEEKS AFTER 6-12 Hr PRESERVATION 10
8
Control
Collins
uw
PHP
LIU ET AL.
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
5 60
clamp, the donor artery and PV were anastomosed in end to side manner to the recipient artery and IVC using continues 8-0 nylon sutures. Then, the recipient’s native small bowel (about 20cm) was resected. Both intestinal ends of grafts were anastomosed to the recipient’s intestine by an end-to- end manner. The intestine anastomosis was performed using a single layer with an interrupted 6-0 silk suture. Postoperative: The rats were kept on a warm condition and under a heating lamp for the first 24 hours. They usually recovered from the anesthesia within 30 minutes after operation. Water was given immediately after operation, The normal water and food were given ad libitum after first 21 hours. However,transfusion was not performed after operation. RESULT The recipient rats transplanted with the grafts after the simple cold-storage and intraluminal perfused preservation by Collins, UW solution and PHP solution for 6 hrs, 12hrs, and 24 hrs were carefully observed. In 6 hr preservation,all of the animals (100%) survived for more than 4 weeks in all group (Fig.1). All of the rats showed decrease in body weight after operation. Some rats were complicated with diarrhea. However,no rats died for this periods. The gain of body weight were observed from 2 weeks after operation in all of rats(Fig.2). The level of serum total protein and albumin had a decline after transplantation, but up to 4 weeks after grafting, they showed recovery to normal (Fig 3 ) . There was no significant difference between the simple cold-storage and intraluminal perfusion preservation. In 12 hr preservation: In group 4 , five of six rats (83%) survived for m o r e than 4 weeks. In all of those, the gain of body weight were observed starting 3 rieeks after grafting. The level of serum total protein and albumin recovered to near normal at 4 weeks after transplantation. Only one of six rats died on seventh day after operation. The gain of body weight was not observed until death. The cause of death was found to be graft necrosis. In group 2, three of six rats survived for more than 4 weeks. In all of those, the gain of body weight was observed and the level of serum total protein and albumin recovered to normal at 4 weeks after operation. Three of six rats(50f) died until 10th day after operation. The causes of death were found to be graft necrosis and arterial thrombosis (n=2),graft necrosis (n=l). weeks after In group 3 , three of six rats survived for more than 4 grafting. The gain of body weight was observed. Recovery of a level of serum total protein and albumin were observed at 4 weeks after transplantation. Three of six rats(50X) were died until 10th day after operat.ion. The cause of death was found to be necrosis in two rats, but in one animal, the cause of death could not be determined. In 24 hr preservation: There was only one of six rats (16%) surviving in each group until 4 weeks after OIT with normal body weight and level of serum protein and albumin. However, other rats in each group survived less than five days after-OIT.
DISCUSSION It was reported that the small intestine was successfully perfused with heparinized lactated Ringer’s solution at normothermia for five hours(9). Utilizing pulsatile perfusion and whole blood at 37*C, the intestine was kept viable in vitro for 18 hours(6). When nonpulsatile flow was applied,,the small bowel was kept viable only for six hours(6). In 1969, Hohenleitner and Senior(lO1 perfused the dog’s entire small bowel with either erythrocyte solution o r high-molecular-weight dextran f o r five hours, and observed indirect signs of viability. Two years later, Alican and associates stored two canine small bowel allografts for 24 hours by continuous hypothermia perfusion with homologous serum at 9%. After orthotopic transplantation, the recipients died within a
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
PYRIDOXYLATED HEMOGLOBIN POLYOXYETHYLENE CONJUGATE SOLUTION
561
few hours due to a loss of fluid, electrolytes, and protein. This phenomenon was interpreted as inadequate small bowel storage(l1). The pressure, vascular resistance and fluid were recognized as acceptable parameters of viability in perfusion storage.Oxygen consumption and enzymatic changes in the perfusate were good indicators of viability during preservation, also. In this study, we performed a model of OIT in the rat in order to investigate whether PHP solution is suitable as a small intestine preservation solution in comparison with Collins and UW solution. We gained 8 3 % survival after 12 hr preservation using PHP solution as a intraluminal perfused solution,with a normal increase of body weight and a normal level of serum protein and albumin 4 weeks after OIT. It was better than other groups.We expect PHP solution to be a new kind of perfusate in intestine preservation.
ACKNOWLEDGEMENT: Pyridoxylated hemoglobin-polyosyethylene is courteously supplied through Ajinomoto Co.,Inc.by a research group of the department of Health and Welfare of Japan. REFERENCES 1.Iwasaki K , Iwashita Y: Preparation and evaluation of hemoglobin-polyethylene glycol conjugate (pyridoxalated polyethleneglycol hemoglobin) as an oxygen carrying resuscitation fluid Artif Organs 10:111-416.1986. 2.Iwasaki li, Iwashita Y , Ikeda k , Llematsu 7 : Efficacy and safety of conjugate (pyridoxnlated polyethylene glycol hemoglobin) as an osygen-carrying resuscitation fluid. Artif Organs 10:470-474.1986. 3,Fuchinoite S, Nakajima I, .Agishi T,et a1:Evaluation of a as a pyridosylated hemoglobin-polyoxyethylene conjugate solution perfusate for liver preservation. Asaio 10: 3. 390-394.1987. 4.Kirkman RL, Lear PA, Madara JL, Tilney NL:Small intestine transplantation in the rat:Immunology and function. Surgery 96:280-287.1981. 5 . L y o r i s GW,et a1:Intestinal absorption of ileum preserved by hypothermia and hyperbaric oxygen.Surg Forum 16:357-359,1965. 6.Aitsten WG, Mclaughlin ED:In vitro small bowel perfusion.Surg Forum 16:359-361,1965. 7.Monchik,G.J.,Russel,P.S:Transplantation of the small bowel in the rat: Technical and immunological consideration. Surgery.50:693-i02,1971. 8 . Wolfgang,H.S. ,Abraham,V.S.,Lee,K.W: Portal versus caval venous drainage of small bowel allografts: Technical and metabolic consequences.Surqery.99:2.193-198,1986.
9.Iijima.K, Salermo RA: Survival of the small intestine following excision,perfusion and autotransplantation. Ann Surg 166:968-975,1967. 10.Hohenleitner FJ, Senior JR: Metabolism of canine small intestine \'ascularly perfused in vitro. J Appl Physiol 26:119-128,1969 11.Alican F , et al: Intestinal transplantation: Laboratory experience and report of a clinical case. Am J Surg 121:150-159,1971
BIOMAT., ART. CELLS & IMMOB. BIOTECH., 20(2-4), 557-561 (1992)
EVALUATION OF A PYRIDOXYLATED HEMOGLOBIN POLYOXYETHYLENE SOLUTION AS A PERFUSATE FOR SMALL INTESTINE PRESERVATION H.I.iit,
CONJUGATE
T.Agishi, T,ha*,ai,T.Hayashi, S.Fu.iita, S.Fuchinoue,li.Takahashi,
s . ' l e r a o k r i 6 h.Ota.
IJcpt .of surgery 1 1 1 , T o k y o k'omen's Yedical ('01 lege.8-1 Kawada-cho, ~ ~ r k u - k uT, o k y o 162, Japan.
Shin
ABSTTtACT new type of artificial blood, pyridoxylated hemoglobinpolgovyethglene conjuqatr( FHP) solution,(developed by PHP research group of the department of health and welfare of' Japan,and produced by Ajinomoto ro.,Inc.Tokyol a s an oxygen-carrying component, has been recently dr\,ised using hemoglobin obtained from hemolyzed human erythrocytes. Recently, the studics using this solution as a preservation sol~ttiortwere> performed in some instances. To r\amine the mechanism of improved ~ i a b i l i t y using this solution u s n preser\'ation solution, we developed a model of orthotopic s m a l l intestine transplantation ( O I T J in the rat. As a baseline study, he comparcd parameters of \lability of the grafts preserved in Collins and L'K soIutioi1 to those preser\ed i i i PHP solution including a survival rate, a st-rum level t o t a l protein and albumin, and a change in body weight after transplantation. T n o u r stiidy, the simplr hypot hermia storage together with intestinal Iwrfirsion preserlation with PHP solution was performed. ,\riimals w e r e divided into 6 , 1 3 , and 2 1 hr preservation groups. 4 1 1 of the r a t s survived after G hr preser\'ation following transplantation. However, in 1 2 hr storage, five of S I Y rats in PHP solution preser\ a t i o n sitr\.i\.eri and r e c o \ e r ) i i t hod? weight after grafting was better that, those kith Collins and L'K solution. k c C O I I C Iude that thr PHF solutiori is,therefore, considered to possit 2 1 > be H more suitable perf1csat.e for small intestine preservation than Collins and I'k solution. KEY WORDS: sinall intestine, transplantation, preservation, perfusion, pyridovglared hemoqlobin polyoxyethglene conjugate solution. ABBREVIATIONS PHP solution:Pyrido\ylnted Hemoglobin Polyoxyethylene solution OIT : 0 1 - t hot op i c in t e s t i ne t ransp 1an tat ion. IVC: lnfrrior vena cave. PV: Portal vein. UW: I'niversity of wisconsin. 4
INTRODUCTION Research and development of artificial blood as a total blood substitute has been carried out by many investigators. PHP conjugate solution as a perfusion solution for organ preservation was reported in 1986 and itsed for l i x - e r preservation( 1-3). The successful use of Cgclosporine .4 in transplantation of other vascularized organ grafts has encouraged ils to use i n small intestine transplantation.(l). However, the preservation time that allows the harvested intestine to remain viable is limited in short time by currently available techniques. If a longer preservation period is allowed for adequate transplantation, patient death due to primary graft failure is supposed to be pre\-ented. 557
Copyright 0 1992 by Marcel Dekker, Inc.
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
5 58
LIU ET AL.
At present, a tested and proven technique for small intestine preservation which is suitable for clinical application is lacking. There is an abundance of conflicting literature on experimental approaches and models of the many techniques for preserving the small intestine ranging from simple hypothermia storage in a hyperbaric oxygen chamber to pulsatile normothermic perfusion.(5-6). Recently PHP conjugate solution (stabilized hemoglobin), which is devised from hemolyzed human erythrocytes,has been developed as an artificial blood with oxygen-carrying property. It was our objective to preserve small intestine by perfusion of the intestinal lumen with 4OC PHP solution. In this study, four groups were divided by intact control(n=6), and Collins solution (n=18), UW solution(n=18), and PHP solution(n=18). The simple storage was performed with Collins and UW solution, and the additional intraluminal perfusion preservation with oxygen supply was performed with PHP solution. In each group, 6, 12, and 24 hr preservation (n=6) was carried out. In the surviving rats, serum level of total protein and albumin, a change of body weight, as parameters for viability of grafts, was checked after transplantation. MATERIALS AND METHODS
Animals, young Lewis strain rats weighting between 250 and 300g, were used both for donor and recipient in order to exclude of the immunologic problem. All of the rats were fed at Tokyo Women's Medical College Animal Experiment Center in temperature-controlled clean cages and received standard laboratory chow and rater ad libitum. Operative technique The rats were fasted for 12 hours before the operation but were allowed free access to water. They were anesthetized with ether. Donor operation: The surgical techniques were modified slightly from those originally described by Monchik and Wolfgang(7-8). The abdomen was opened a midline incision. A microscope was used to mobilize the graft. Manipulation of the intestine was tried to be minimal. An about 20cm segment of donor small intestine,the proximal jejunum apart 5cm from the ligament of Treitz, was isolated on a vascular pedicle consisting of the superior mesenteric artery with a segment of the aorta and the portal vein. The lumbar arteries from the aorta were meticulously ligated with 6-0 silk suture to minimize bleeding. After systemic heparization (15Ou/lOOg), the aorta was cannulated, and the graft vascular bed was perfused with 5-101111of heparinized lactated Ringer's solution. A small volume of cold saline solution containing gentamycin was used to irrigate the gut lumen avoiding overdistention of the donor bowel. The portal vein and the aorta above and below the superior mesenteric artery was transected with an aorta cuff, then the graft was removed. The harvested intestine was then preserved in three groups: Group 1: Control group (11.61 consisting of age and weightmatched Lewis strain rats in which no surgical procedures were performed. 2 : Collins solution: The graft vascular bed and intestinal were Qroup perfused with Collins solution and preserved for 6 hrs(n=6), 12 hrs(n=6), 24 hrs(n=6) at 4%. 3 : tJW solution: The graft vascular bed and intestinal were Qroup perfused with UW solution and preserved for 6 hrs(n=6),12 hrs(n.61, 24 hrs(n.6) at 4OC. 4: PHP solution: The graft vascular bed was Group perfused with PHP solution. Subsequently, a short wide rubber tube was inserted into the bowel for continuous perfusion preservation ( speed: 2Oml/hr )with oxygen for 6 hrs, l2hrs, and 24 hrs at 4%. Recipient operation: Recipient animals were generally anesthetized similarly to donor. The segment grafts were placed orthotopically. Revascularization was performed between the donor and recipient. The aorta and inferior vena cave were exposed, by means of a modified Lee's
559
PYRIWXYLATED HEMOGLOBIN POLYOXYETHYLENE CONJUGATE SOLUTION
I SMALL INTESTINAL PRESERVATION I (0
I1 6
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
C
H5 5 4
w
A 3
.g
x
2
czl 0
PHP
Control
Collins
(Fig. 1 )
POSTOPERATIVE BODY WEIGHT CHANGE FOLLOWING 6 Hr PRESERVATION I 0
0
1
2
3
4
WEEKS
(Fig. 2 )
SERUM TOTAL PROTEIN AND ALBUMIN CONCENTRATION 4 WEEKS AFTER 6-12 Hr PRESERVATION 10
8
Control
Collins
uw
PHP
LIU ET AL.
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
5 60
clamp, the donor artery and PV were anastomosed in end to side manner to the recipient artery and IVC using continues 8-0 nylon sutures. Then, the recipient’s native small bowel (about 20cm) was resected. Both intestinal ends of grafts were anastomosed to the recipient’s intestine by an end-to- end manner. The intestine anastomosis was performed using a single layer with an interrupted 6-0 silk suture. Postoperative: The rats were kept on a warm condition and under a heating lamp for the first 24 hours. They usually recovered from the anesthesia within 30 minutes after operation. Water was given immediately after operation, The normal water and food were given ad libitum after first 21 hours. However,transfusion was not performed after operation. RESULT The recipient rats transplanted with the grafts after the simple cold-storage and intraluminal perfused preservation by Collins, UW solution and PHP solution for 6 hrs, 12hrs, and 24 hrs were carefully observed. In 6 hr preservation,all of the animals (100%) survived for more than 4 weeks in all group (Fig.1). All of the rats showed decrease in body weight after operation. Some rats were complicated with diarrhea. However,no rats died for this periods. The gain of body weight were observed from 2 weeks after operation in all of rats(Fig.2). The level of serum total protein and albumin had a decline after transplantation, but up to 4 weeks after grafting, they showed recovery to normal (Fig 3 ) . There was no significant difference between the simple cold-storage and intraluminal perfusion preservation. In 12 hr preservation: In group 4 , five of six rats (83%) survived for m o r e than 4 weeks. In all of those, the gain of body weight were observed starting 3 rieeks after grafting. The level of serum total protein and albumin recovered to near normal at 4 weeks after transplantation. Only one of six rats died on seventh day after operation. The gain of body weight was not observed until death. The cause of death was found to be graft necrosis. In group 2, three of six rats survived for more than 4 weeks. In all of those, the gain of body weight was observed and the level of serum total protein and albumin recovered to normal at 4 weeks after operation. Three of six rats(50f) died until 10th day after operation. The causes of death were found to be graft necrosis and arterial thrombosis (n=2),graft necrosis (n=l). weeks after In group 3 , three of six rats survived for more than 4 grafting. The gain of body weight was observed. Recovery of a level of serum total protein and albumin were observed at 4 weeks after transplantation. Three of six rats(50X) were died until 10th day after operat.ion. The cause of death was found to be necrosis in two rats, but in one animal, the cause of death could not be determined. In 24 hr preservation: There was only one of six rats (16%) surviving in each group until 4 weeks after OIT with normal body weight and level of serum protein and albumin. However, other rats in each group survived less than five days after-OIT.
DISCUSSION It was reported that the small intestine was successfully perfused with heparinized lactated Ringer’s solution at normothermia for five hours(9). Utilizing pulsatile perfusion and whole blood at 37*C, the intestine was kept viable in vitro for 18 hours(6). When nonpulsatile flow was applied,,the small bowel was kept viable only for six hours(6). In 1969, Hohenleitner and Senior(lO1 perfused the dog’s entire small bowel with either erythrocyte solution o r high-molecular-weight dextran f o r five hours, and observed indirect signs of viability. Two years later, Alican and associates stored two canine small bowel allografts for 24 hours by continuous hypothermia perfusion with homologous serum at 9%. After orthotopic transplantation, the recipients died within a
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Sydney on 05/02/15 For personal use only.
PYRIDOXYLATED HEMOGLOBIN POLYOXYETHYLENE CONJUGATE SOLUTION
561
few hours due to a loss of fluid, electrolytes, and protein. This phenomenon was interpreted as inadequate small bowel storage(l1). The pressure, vascular resistance and fluid were recognized as acceptable parameters of viability in perfusion storage.Oxygen consumption and enzymatic changes in the perfusate were good indicators of viability during preservation, also. In this study, we performed a model of OIT in the rat in order to investigate whether PHP solution is suitable as a small intestine preservation solution in comparison with Collins and UW solution. We gained 8 3 % survival after 12 hr preservation using PHP solution as a intraluminal perfused solution,with a normal increase of body weight and a normal level of serum protein and albumin 4 weeks after OIT. It was better than other groups.We expect PHP solution to be a new kind of perfusate in intestine preservation.
ACKNOWLEDGEMENT: Pyridoxylated hemoglobin-polyosyethylene is courteously supplied through Ajinomoto Co.,Inc.by a research group of the department of Health and Welfare of Japan. REFERENCES 1.Iwasaki K , Iwashita Y: Preparation and evaluation of hemoglobin-polyethylene glycol conjugate (pyridoxalated polyethleneglycol hemoglobin) as an oxygen carrying resuscitation fluid Artif Organs 10:111-416.1986. 2.Iwasaki li, Iwashita Y , Ikeda k , Llematsu 7 : Efficacy and safety of conjugate (pyridoxnlated polyethylene glycol hemoglobin) as an osygen-carrying resuscitation fluid. Artif Organs 10:470-474.1986. 3,Fuchinoite S, Nakajima I, .Agishi T,et a1:Evaluation of a as a pyridosylated hemoglobin-polyoxyethylene conjugate solution perfusate for liver preservation. Asaio 10: 3. 390-394.1987. 4.Kirkman RL, Lear PA, Madara JL, Tilney NL:Small intestine transplantation in the rat:Immunology and function. Surgery 96:280-287.1981. 5 . L y o r i s GW,et a1:Intestinal absorption of ileum preserved by hypothermia and hyperbaric oxygen.Surg Forum 16:357-359,1965. 6.Aitsten WG, Mclaughlin ED:In vitro small bowel perfusion.Surg Forum 16:359-361,1965. 7.Monchik,G.J.,Russel,P.S:Transplantation of the small bowel in the rat: Technical and immunological consideration. Surgery.50:693-i02,1971. 8 . Wolfgang,H.S. ,Abraham,V.S.,Lee,K.W: Portal versus caval venous drainage of small bowel allografts: Technical and metabolic consequences.Surqery.99:2.193-198,1986.
9.Iijima.K, Salermo RA: Survival of the small intestine following excision,perfusion and autotransplantation. Ann Surg 166:968-975,1967. 10.Hohenleitner FJ, Senior JR: Metabolism of canine small intestine \'ascularly perfused in vitro. J Appl Physiol 26:119-128,1969 11.Alican F , et al: Intestinal transplantation: Laboratory experience and report of a clinical case. Am J Surg 121:150-159,1971
