Teemu Lämsä

Tissue adhesives and the pancreas: biocompatibility and adhesive properties of 6 preparations.

Objectives: The aim of this study was to investigate biocompatibility and adhesive properties of 6 tissue adhesives available, when applied between the pancreas and jejunum in an experimental model. Methods: Portion of jejunum was glued on the pancreas in rats with 3 cyanoacrylate derivatives (Histoacryl, Dermabond, and Glubran 2), 2 human fibrin sealants (Tisseel Duo Quick and Quixil), and 1 albuminglutaraldehyde sealant (BioGlue). Pancreatic tissue specimens and blood samples were harvested 1, 3, 7, and 21 days after gluing for histological determination and amylase activity measurement. Pancreaticojejunal attachment created with adhesives underwent tensile strength measurement at each time point. Samples were also taken from unoperated rats and sham-operated rats. Results: Exposure and sham groups both induced a similar increase in amylase activity on day 1 with normalization by day 3. Sham operation induced mild changes in the pancreas. Each tissue adhesive induced changes in pancreatic histology to the entire gland. Injurious effect was more severe with the 3 cyanoacrylates than with the 3 fibrin/semisynthetic glues. Histoacryl and Quixil induced lower tensile strength than the other adhesives. Conclusions: All of the tissue adhesives studied induced histological changes in the pancreas of which at least part might be considered harmful. The potentially harmful tissue effects of the preparations tested might compromise the use of these substances in pancreatic surgery.

Biocompatibility of a New Bioabsorbable Radiopaque Stent Material (BaSO4 Containing Poly-L,D-Lactide) in the Rat Pancreas

Background/Aim: During recent years, we have been developing bioabsorbable biliary stents with promising experimental results. In developing pancreatic stents before long-term experiments, the acute toxicity to the pancreas of a bioabsorbable, radiopaque polylactide (PLA 96-barium sulfate, BaSO4) stent material was investigated. Methods: The pancreas of 65 Sprague-Dawley rats was exposed either to radiopaque stent material [PLA 96 with 25% (w/w) of BaSO4], radiolucent stent material (PLA 96), or inert steel by inserting a 5-mm-long (diameter 0.3 mm) fiber/stick of material into the pancreas after laparotomy under general anesthesia. Pancreatic tissue specimens and blood samples were taken after 1, 3, 7, and 21 days for histological examination and amylase activity measurements. Samples were also taken from 5 baseline (control) rats without exposing to any materials. Results: The baseline serum amylase activity was normal, and no histological changes in the pancreas were observed. A significant increase (mean ± SE) in the serum amylase activity was observed only on day 1 in the animals having radiopaque stent material (PLA 96-BaSO4; 5,845 ± 1,135 U/l), steel (4,946 ± 667 U/l), or radiolucent stent material (PLA 96; 7,684 ± 667 U/l) inserted. There was slightly more acinar cell necrosis on day 7 in the steel group than in the radiopaque stent (PLA 96-BaSO4) group (p = 0.028). Conclusions: Radiopaque stent material (PLA 96-BaSO4) was not more toxic than the reference steel material in the rat pancreas during the 21-day observation period and is thus applicable for further in vivo experiments when developing pancreatic bioabsorbable stents.

A novel biodegradable pancreatic stent for human pancreatic applications: a preclinical safety study in a large animal model

BACKGROUND Endoscopic stenting is one treatment method for pancreatic strictures or pseudocysts in patients with symptomatic chronic pancreatitis. With a biodegradable stent, the later removal of the stent could be avoided. OBJECTIVE We investigated the degradation, patency, and toxicity of a novel biodegradable, self-expanding radiopaque polylactide-barium sulfate pancreatic stent in a large animal model. DESIGN Animal study. SETTING AND INTERVENTIONS Five swine had a biodegradable pancreatic stent placed into the pancreatic duct (PD) and were followed-up for 6 months. MAIN OUTCOME MEASUREMENTS Repeated blood tests and radiographs were studied during the follow-up. The animals were euthanized at 6 months, at which time, the PD inner diameter was measured, and histology was analyzed and graded. For comparison, histology from 5 nonstented animals was also analyzed. RESULTS The stent was correctly inserted into the PD in 4 of 5 animals. All the animals remained healthy, gained weight, and showed no signs of pancreatitis. A radiograph showed that the stent was in its original place in all animals at 1 month but had disappeared in all animals by 3 months. At 6 months, the autopsied pancreatic tissue was soft, and the PD was patent in all of the animals. The PD was slightly dilated at the site of the stent in the head of the pancreas compared with the preoperative diameter (5 mm [range 3-6 mm] vs 2 mm [range 1-3 mm], P < .05), but, in the body of the pancreas, no significant dilatation was seen. In the histology of the pancreata, there was no difference between the samples from the exposed parts of the biodegradable pancreatic stent compared with the distal nonexposed parts, or to the samples from the nonstented animals. CONCLUSIONS This novel biodegradable pancreatic stent, studied in these swine, appeared to be safe for use in the PD. These encouraging results warrant further clinical trials with this biodegradable pancreatic stent in pancreatic applications in human beings.

Pancreatic Injury Response is Different Depending on the Method of Resecting the Parenchyma

OBJECTIVES The present study was performed to compare the pancreatic injury response on the parenchymal resection either with ultrasonic scissors, electrocautery, or surgical scalpel. METHODS A 1 x 0.5 cm piece of rat pancreas was resected from side of the pancreas either with ultrasonic scissors (Harmonic Scalpel; UltraCision, Ethicon Endosurgery Inc., Cincinnati, OH) or electrocautery (Force FX; Valleylab, Tyco Healthcare Group LP, Boulder, CO) at two power levels, 1 and 3; 8W and 25W, respectively, or with surgical scalpel. Hemostasis was provided after surgical scalpel either with cellulose patch (Interceed; Johnson and Johnson Medical, Inc., New Brunswick, NJ), three stitches of 6-0 polydioxanone at tightness of 0.6N or fibrin glue (Tisseel Duo Quick; Baxter AG, Wien, Austria). Blood sample and pancreas specimens, both at the resection site and far away, were taken 1, 7, and 21 days postoperatively from exposed animals, sham operated animals (n = 18 in each) and from unexposed baseline animals (n = 5). Necrosis, edema, leukocyte infiltration, hemorrhage, vacuolization, and fibrosis were histologically assessed separately. RESULTS Each resection and sham operation induced similar increase in the amylase activity on day 1 with normalization by day 7. Resection with ultrasonic scissors and electrocautery induced more tissue injury to the pancreas than resection with surgical scalpel independent of the method for hemostasis. The injury, although somewhat milder in intensity, was also observed in parts of the pancreas located far away from the site of resection. CONCLUSIONS Of the compared methods, surgical scalpel resection plus cellulose patch or fibrin glue hemostasis induced the least histological changes in the pancreatic parenchyma. This injury response spread over the pancreas.

A polyamine analog bismethylspermine ameliorates severe pancreatitis induced by intraductal infusion of taurodeoxycholate.

BACKGROUND Stable polyamine homeostasis is important for cell survival and regeneration. Our experimental studies have shown that catabolism of spermidine and spermine to putrescine is associated with the development of pancreatitis. We investigated the pathogenetic role of polyamine catabolism by studying the effect of a methylated polyamine analog on taurodeoxycholate-induced acute experimental pancreatitis. METHODS Acute pancreatitis was induced by infusion of sodium taurodeoxycholate (2%) into the pancreatic duct. Bismethylspermine (Me(2)Spm) was administered as a pretreatment before the induction of pancreatitis or as a treatment after the induction of pancreatitis. The sham operation included laparotomy only. Pancreas tissue and blood were sampled at 24 h and 72 h after the infusion of taurodeoxycholate and studied for pancreatitis severity (serum amylase activity, pancreatic water content, and histology) and polyamine catabolism, which includes spermidine/spermine N(1)-acetyltransferase (SSAT) activity as well as spermidine, spermine, and putrescine concentrations in the pancreas. RESULTS Sodium taurodeoxycholate-induced acute pancreatitis manifests as increases in serum amylase and pancreatic water content, leukocytosis, and acinar cell necrosis in the pancreas. The activity of SSAT increased significantly together with an increase in the ratios of pancreatic putrescine/spermidine and putrescine/spermine at 24 h, which indicates SSAT-induced polyamine catabolism. Pancreatic water content and necrosis were reduced significantly by the treatment with Me(2)Spm at 24 h but not at 72 h when the polyamine homeostasis had recovered, and the pancreatitis had progressed. CONCLUSIONS Taurodeoxycholate-induced acute pancreatitis was associated with activation of polyamine catabolism in the pancreas. The polyamine analog Me(2)Spm ameliorated the injury in the early stage, but it did not ameliorate the late progression of the pancreatic necrosis at 72 h. Thus, besides proteolytic enzyme activation and the cascades of inflammation, polyamine catabolism may be an important pathogenetic mediator of the early stages of acute pancreatitis.

Polyamine Levels in the Pancreas and the Blood Change according to the Severity of Pancreatitis

Background/Aims: Polyamines are essential to survival, growth, and proliferation of mammalian cells. Previous studies have suggested that the pancreatic polyamine levels may change in acute pancreatitis. In this study, the changes of polyamine levels in the pancreas have been studied with respect to the severity of pancreatitis. We investigated whether there is a relationship in polyamine levels between pancreas and blood, and whether pancreatic and blood polyamine levels change according to the severity of pancreatitis. Methods: In rats, sublethal pancreatitis was induced by intraductal infusion of 2% taurodeoxycholate, while lethal pancreatitis was induced with 6% taurodeoxycholate. Results: Infusion of 6% taurodeoxycholate as compared with 2% resulted in more severe pancreatitis, as revealed by mortality, histology, and serum amylase activity. Pancreatic spermidine/spermine N1-acetyltransferase was induced early after pancreatitis and was associated with increased putrescine and decreased spermidine levels. The extent of pancreatic necrosis significantly correlated with the polyamine catabolism indicators pancreatic putrescine/spermidine ratio (r = 0.29, p < 0.01) and pancreatic putrescine/spermine ratio (r = 0.32, p < 0.01). The two pancreatic polyamine ratios correlated well also with the red blood cell polyamine ratios (r = 0.75 and r = 0.72, respectively, both p < 0.01). Furthermore, the extent of pancreatic necrosis correlated with red blood cell putrescine/spermidine (r = 0.32, p < 0.01) and putrescine/spermine (r = 0.37, p < 0.01) ratios. Conclusions: Acute experimental pancreatitis is associated with an early pancreatic spermidine/spermine N1-acetyltransferase induction and consequent changes in polyamine levels in pancreas and red blood cells, depending on the severity of pancreatitis. Because changes in red blood cell spermidine, spermine, and putrescine levels evolve already early during the time course of pancreatitis, and correlate with the extent of pancreatic necrosis, their clinical value as early markers of the severity of acute pancreatitis needs to be further evaluated.

Effects of Diameter, Number and Tightness of Sutures on Pancreatic Injury Response

Background: We have experimental data indicating that the pancreas is easily damaged by any intervention. The present study compared the effects of suture diameter, number of needle passes and suture tightness on rat pancreas. Methods: Under anesthesia, rat pancreas was sutured either with one loose stitch of 6-0 polydioxanone (PDS II) or 3-0 PDS II, with 5 passes of loose running 6-0 PDS II, or with 6-0 PDS II loop tightened to 0.6 or 1.2 N. Amylase activity and pancreatic tissue histology at the suturing site and farther away, were evaluated 1, 3, 7 and 21 days postoperatively. Results: Each suturing exposure and the sham-operation induced temporary amylase activity elevation on day 1 when compared with the baseline. In histology, 3-0 suture, 5 needle passes and 1.2-newton loop induced more damage than 6-0 suture, single needle pass and 0.6-newton loop, respectively. Similar but milder changes were observed in samples from the remote site. Conclusions: The pancreas reacts to suturing with widespread injury response resembling that of acute pancreatitis. In attempting to reduce suturing-induced widespread injury, as few and thin sutures and as loose suture tightness as possible should be used. Although these findings may seem obvious, they have not previously been proven in terms of histology.

Polyamine Catabolism in Relation to Trypsin Activation and Apoptosis in Experimental Acute Pancreatitis

Background: Overinduced polyamine catabolism (PC) in a transgenic rat model has been suggested to be a mediator of trypsin activation which is important in acinar cell necrosis. PC has also been observed in experimental taurodeoxycholate pancreatitis. We hypothesized that PC may be a mediator of trypsin activation in taurodeoxycholate pancreatitis. Methods: Pancreatitis was induced in wild-type rats by 2 or 6% taurodeoxycholate infusion or in transgenic rats by overexpressing spermidine/spermine N1-acetyltransferase (SSAT). The time courses of necrosis, caspase-3 immunostaining, SSAT, polyamine levels, and trypsinogen activation peptide (TAP) were monitored. The effect of the polyamine analogue bismethylspermine (Me2Spm) was investigated. Results: In a transgenic pancreatitis model, TAP and acinar necrosis increased simultaneously after the activation of SSAT, depletion of spermidine, and development of apoptosis. In taurodeoxycholate pancreatitis, necrosis developed along with the accumulation of TAP. SSAT was activated simultaneously or after TAP accumulation and less than in the transgenic model, with less depletion of spermidine than in the transgenic model. Supplementation with Me2Spm ameliorated the extent of acinar necrosis at 24 h, but contrary to previous findings in the transgenic model, in the taurodeoxycholate model it did not affect trypsin activation. Compared with the transgenic model, no extensive apoptosis was found in taurodeoxycholate pancreatitis. Conclusions: Contrary to transgenic SSAT-overinduced pancreatitis, PC may not be a mediator of trypsin activation in taurodeoxycholate pancreatitis. The beneficial effect of polyamine supplementation on necrosis in taurodeoxycholate pancreatitis may rather be mediated by other mechanisms than amelioration of trypsin activation.

Association between remote organ injury and tissue polyamine homeostasis in acute experimental pancreatitis – treatment with a polyamine analogue bismethylspermine

Experimental pancreatitis is associated with activation of polyamine catabolism. The polyamine analog bismethylspermine (Me(2)Spm) can ameliorate pancreatic injury. We investigated the roles of polyamine catabolism in remote organs during pancreatitis and explored the mechanism of polyamine catabolism by administering Me(2)Spm. Acute pancreatitis was induced by an infusion of 2 or 6% taurodeoxycholate before Me(2)Spm administration. Blood, urine and tissues were sampled at 24 and 72 h to assess multi-organ injury and polyamine catabolism. The effect of Me(2)Spm on mortality in experimental pancreatitis was tested separately. Liver putrescine levels were elevated following liver injury. Me(2)Spm increased the activity of spermidine/spermine N(1)-acetyltransferase (SSAT) and depleted the spermidine, spermine or putrescine levels. Lung putrescine levels increased, and SSAT and spermine decreased following lung injury. Me(2)Spm enhanced the activity of SSAT and decreased the spermidine and spermine levels. Renal injury was manifested as an increase in creatinine or a decrease in urine output. Decreases in kidney SSAT, spermidine or spermine and an increase in putrescine were found during pancreatitis. In the 2% taurodeoxycholate model, Me(2)Spm decreased urine output and raised plasma creatinine levels. Me(2)Spm increased SSAT and decreased polyamines. Excessive Me(2)Spm accumulated in the kidney, and greater amounts were found in the 6% taurodeoxycholate model in which this mortality was not reduced by Me(2)Spm. In the 2% taurodeoxycholate model, Me(2)Spm dose-dependently induced mortality at 72 h. Like pancreatic injury, remote organ injury in pancreatitis is associated with increased putrescine levels. However, Me(2)Spm could not ameliorate multi-organ injury. Me(2)Spm administration was associated with significant renal toxicity and induced mortality, suggesting that the current dose is too high and needs to be modified.