Fred Bonthuis

Reduction of peritoneal trauma by using nonsurgical gauze leads to less implantation metastasis of spilled tumor cells

OBJECTIVES To evaluate whether infliction of peritoneal trauma would promote tumor cell adherence to damaged peritoneal surfaces; to investigate whether peritoneal damage could promote tumor growth of extraperitoneal tumors; and to evaluate whether the amount of trauma correlated with the degree of tumor cell adherence and local and distant tumor growth. BACKGROUND DATA After potentially curative resection of colorectal carcinoma, the most common site for recurrence is locoregional. We previously demonstrated that surgical trauma induces a cascade of events leading to adhesion formation. The same mechanisms may be responsible for improved tumor cell adherence and growth facilitation in early local recurrence. METHODS A reproducible rat model was used in which peritoneal damage was inflicted by standardized rubbing of the peritoneum with surgical gauzes of different texture. In the first experiment, tumor cell adherence and growth at traumatized and nontraumatized peritoneal sites were assessed semiquantitatively 3 weeks after perioperative intra-abdominal injection of CC-531 tumor cells. In the second experiment, the effect of peritoneal trauma on ectopic tumor growth was investigated (CC-531 implanted under the renal capsule). In the final experiment, we evaluated how soon after peritoneal traumatization tumor cell adhesion and growth-promoting factors were active and whether they could be passively transferred to naïve nontraumatized abdominal cavities. RESULTS A significant correlation between the amount of peritoneal trauma and the degree of tumor take at damaged peritoneal surfaces was found (p < or = 0.018). Tumor take at remote peritoneal sites not directly traumatized was also significantly higher after severe trauma than after moderate trauma of the peritoneum (p < or = 0.005). In addition, a significant correlation between the degree of peritoneal trauma and the growth of ectopic tumors under the renal capsule was observed (p < or = 0.009). The final experiment demonstrated that within a few hours after infliction of peritoneal trauma, tumor growth-promoting effects could be passively transferred to naïve recipients. CONCLUSIONS Surgical trauma is an important factor in the promotion of local recurrence. The enhancing effect of trauma is not restricted to the inflicted site but rather has a generalized character. Avoidance of unnecessary surgical trauma by using gentle techniques and materials is therefore indicated.

Prevention of adhesion formation to polypropylene mesh by collagen coating: A randomized controlled study in a rat model of ventral hernia repair

IntroductionIn laparoscopic incisional hernia repair with intraperitoneal mesh, concern exists about the development of adhesions between bowel and mesh, predisposing to intestinal obstruction and enterocutaneous fistulas. The aim of this study was to assess whether the addition of a collagen coating on the visceral side of a polypropylene mesh can prevent adhesion formation to the mesh.MethodIn 58 rats, a defect in the muscular abdominal wall was created, and a mesh was fixed intraperitoneally to cover the defect. Rats were divided in two groups; polypropylene mesh (control group) and polypropylene mesh with collagen coating (Parieten mesh). Seven and 30 days postoperatively, adhesions and amount and strength of mesh incorporation were assessed. Wound healing was studied by microscopy.ResultsWith Parieten mesh, the mesh surface covered by adhesions was reduced after 30 days (42% vs 69%, p = 0.01), but infection rate was increased after both 7 (p = 0.001) and 30 days (p = 0.03), compared to the polypropylene group with no mesh infections. If animals with mesh infection were excluded in the analysis, the mesh surface covered by adhesions was reduced after 7 days (21% vs 76%, p = 0.02), as well as after 30 days (21 vs 69%, p < 0.001). Percentage of mesh incorporation was comparable in both groups. Mean tensile strength of mesh incorporation after 30 days was higher with Parieten mesh.ConclusionAlthough the coated Parieten mesh was more susceptible to mesh infection in the current model, a significant reduction of adhesion formation was still seen with the Parieten mesh after 30 days, with comparable mesh incorporation in the abdominal wall.

Long-Term Impact of Pneumoperitoneum Used for Laparoscopic Donor Nephrectomy on Renal Function and Histomorphology in Donor and Recipient Rats

ObjectiveTo investigate the long-term impact of pneumoperitoneum used for laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient. Summary Background DataLaparoscopic donor nephrectomy has the potential to increase the number of living kidney donations by reducing donor morbidity. However, function of laparoscopically procured kidneys might be at risk due to ischemia as a consequence of elevated intra-abdominal pressure during laparoscopy. MethodsIn experiment 1, 30 Brown Norway rats were randomized to three procedures: 2 hours of CO2 insufflation, 2 hours of helium insufflation, and 2 hours of gasless laparoscopy. After this, a unilateral nephrectomy was performed in all animals. Another six rats were used as controls. In experiment 2, 36 donor Brown Norway rats were subjected to a similar insufflation protocol, but after nephrectomy a syngeneic renal transplantation was performed. All rats had a follow-up period of 12 months. Urine and blood samples were collected each month for determination of renal function. After 1 year, donor and recipient kidneys were removed for histomorphologic and immunohistochemical analysis. ResultsIn donors as well as in recipients, no significant changes in serum creatinine, proteinuria, or glomerular filtration rate were detected between the CO2, the helium, and the gasless control group after 1 year. No histologic abnormalities due to abdominal gas insufflation were found. Immunohistochemical analysis did not show significant differences in the number of infiltrating cells (CD4, CD8, ED1, OX62, and OX6) and adhesion molecule expression (ICAM-1) between the three groups. ConclusionsAbdominal gas insufflation does not impair renal function in the donor 1 year after LDN. One year after transplantation, no differences in renal function or histomorphology were detected between kidney grafts exposed to either pneumoperitoneum or a gasless procedure.

A membrane cofactor protein transgenic mouse model for the study of discordant xenograft rejection

Background: In recent years, interest has been revived in the possibility of transplanting organs into humans from a phylogenetically disparate species such as the pig (xenotransplantation). Such discordant xenografts, however, are subject to hyperacute rejection (HAR) and activation of host complement plays a major role in this rejection. This problem may be solved through the use of transgenic technology by providing the grafted tissue with molecules that down‐regulate the action of host complement.

Accumulation of plasma triiodothyronine sulfate in rats treated with propylthiouracil.

Triiodothyronine sulfate (T3S) is rapidly deiodinated by the propylthiouracil (PTU)-sensitive type I deiodinase. Here we examined the effects of PTU on plasma T3S levels in rats after intravenous administration of radiolabeled T3 or T3S. Sephadex LH-20 chromatography and high-performance liquid chromatography were used to quantify conjugated and nonconjugated iodothyronines, and iodide was measured as the TCA-soluble radioactivity. In control rats, radioiodide was the main metabolite of both T3 and T3S. Plasma T3S was cleared more rapidly than plasma T3 despite increased binding to plasma proteins. PTU reduced plasma iodide levels by 66 and 78% after T3 and T3S, respectively, and decreased plasma clearance of T3S by 81%. However, PTU had no effect on plasma T3 clearance but increased plasma T3S from injected T3 4.2 times. Biliary excretion of injected T3S was less than 20% in normal rats, in contrast to 70% within 4 h in PTU-treated rats. In conclusion, T3S is an important intermediate in the in vivo metabolism of T3 in rats and accumulates in plasma if type I deiodination is inhibited.

Synthetic oligopeptides related to the β-subunit of human chorionic gonadotropin attenuate inflammation and liver damage after (Trauma) hemorrhagic shock and resuscitation

Severe hemorrhagic shock (HS) followed by resuscitation induces a massive inflammatory response, which may culminate into systemic inflammatory response syndrome, multiple organ dysfunction syndrome, and, finally, death. Treatments that effectively prevent this inflammation are limited so far. In a previous study, we demonstrated that synthetic oligopeptides related to the primary structure of human chorionic gonadotropin (HCG) can inhibit the inflammatory response and mortality that follow high-dose LPS-induced inflammation. Considering this powerful anti-inflammatory effect, we investigated whether administration of similar synthetic HCG-related oligopeptides (LQGV, AQGV, LAGV) during HS were able to attenuate the inflammatory response associated with this condition. Hemorrhagic shock was induced in rats for 60 min by blood withdrawal until a MAP of 40 mmHg was reached. Rats received a single injection with one of the hCG-related oligopeptides (LQGV, AQGV or LAGV) or 0.9% NaCl solution as control 30 min after induction of HS. Treatment with LQGV, AQGV, or LAGV prevented systemic release of TNF-&agr; and IL-6 and was associated with reduced TNF-&agr;, IL-6, and E-selectin mRNA transcript levels in the liver. LQGV treatment prevented neutrophil infiltration into the liver and was associated with reduced liver damage. Our data suggest that HCG-related oligopeptides, in particular LQGV, have therapeutic potential by attenuating the life-threatening inflammation and organ damage that is associated with (trauma) HS and resuscitation.

Short-term impact of carbon dioxide, helium, and gasless laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient

Background: Laparoscopic donor nephrectomy has the potential to increase the number of living kidney donations by reducing donor morbidity. However, studies have shown that raised intraabdominal pressure can result in transient renal dysfunction. Therefore, laparoscopically procured kidneys might be at higher risk for suffering a period of ischemia during pneumoperitoneum. The objective of this study was to investigate the short-term impact of pneumoperitoneum used for laparoscopic donor nephrectomy on renal function and histomorphology in donor and recipient. Methods: Experiment 1: Kidney donor: Initially, 36 brown Norway (BN) rats were randomized for three procedures: 2 h of carbon dioxide (CO2) insufflation (8 mmHg), 2 h of helium insufflation (8 mmHg), and 2 h of gasless technique (0 mmHg). After this, a unilateral nephrectomy was performed in all the animals. Experiment 2: Recipient: Subsequently, 36 donor BN rats were subjected to a similar insufflation protocol, but after nephrectomy, a syngeneic kidney transplantation (BN-BN) was performed. Urine and blood samples were collected on postoperative days 1, 3, 7, and 14 for determination of renal function. Subsequently, donor and recipient kidneys were removed for histomorphologic and immunohistochemical analysis. Results: In both donors and recipients, no significant changes in serum creatinine, proteinuria, or glomular filtration were detected between the CO2, the helium, and the gasless control groups. In both experiments, histologic analysis of Kidney specimens did not show any deleterious effects from abdominal gas insufflation. Although kidney grafts exposed to CO2 showed significantly higher numbers of CD45+ leukocytes 3 days after transplantation, immunohistochemical analysis did not show significant differences in number of infiltrating cells (CD4, CD8, ED1, OX6, OX62) between the two insufflation groups and the gasless control subjects. Conclusions: Abdominal gas insufflation does not have an adverse effect on the renal function of the kidney donor 1 week after laparoscopic donor nephrectomy. No differences in renal function or histomorphology were detected between syngeneic kidney grafts exposed to pneumoperitoneum and gasless control subjects.

Effects of thyroid status and thyrostatic drugs on hepatic glucuronidation of lodothyronines and other substrates in rats : Induction of phenol UDP-glucuronyltransferase by methimazole.

Glucuronidation of iodothyronines in rat liver is catalyzed by at least three UDP-glucuronyltransferases (UGTs): bilirubin UGT, phenol UGT, and androsterone UGT. Bilirubin and phenol UGT activities are regulated by thyroid hormone, but the effect of thyroid status on hepatic glucuronidation of iodothyronines is unknown. We examined the effects of hypothyroidism induced by treatment of rats with propylthiouracil (PTU) or methimazole (MMI) or by thyroidectomy as well as the effects of T4-induced hyperthyroidism on the hepatic UGT activities for T4, T3, bilirubin,p-nitrophenol (PNP), and androsterone. Bilirubin UGT activity was increased in MMI- or PTU-induced hypothyroid and thyroidectomized rats, and decreased in hyperthyroid animals. T4 and, to a lesser extent, T3 UGT activities were increased in MMI- or PTU-induced hypothyroid rats, and T4 but not T3 glucuronidation also showed a significant increase in thyroidectomized rats. T4 but not T3 UGT activity was slightly decreased in hyperthyroid rats. While PNP UGT activity was decreased in thyroidectomized rats and increased in hyperthyroid animals, it was also markedly increased by MMI and slightly increased by PTU-induced hypothyroidism. In T4-substituted rats, MMI did not affect T4, T3, bilirubin and androsterone UGT activities but again strongly induced PNP UGT activity, indicating that this represented a direct induction of PNP UGT by the drug independent of its thyrostatic action. Androsterone UGT activity was hardly affected by thyroid status. Our results suggest a modest, negative control of the hepatic glucuronidation of thyroid hormone by thyroid status, which may be mediated by changes in bilirubin UGT activity. To our knowledge, this is the first report of the marked induction of a hepatic enzyme by MMI, which is not mediated by its thyroid hormone-lowering effect.

Hyaluronate-based coating solution for prevention of surgical adhesions has no major effect on adhesion and growth of intraperitoneal tumour cells

OBJECTIVE To find out whether the perioperative use of a solution containing hyaluronic acid (HA, Sepracoat) might affect the adhesion of tumour cells. DESIGN Laboratory studies in vitro and in two experiments in rats. SETTING Teaching hospital, The Netherlands. SUBJECTS 27 female inbred WAG rats. INTERVENTIONS Mesothelial cells were cultured in monolayers and the adhesion of CC-531 colonic carcinoma cells was assessed with and without Sepracoat. Uterine horn experiment: after laparotomy Sepracoat 3 ml (n = 5) or phosphate buffered saline (PBS) (n = 4) were instilled in rats; the right uterine horn was abraided with gauze, and the left was left untouched; CC-531 cells were seeded intraperitoneally; and the tumour load at 8 different sites was scored after 3 weeks. Laparotomy model: after laparotomy Sepracoat and PBS were instilled (n = 9 rats in each group), CC-531 cells were seeded, and the wound was closed; the tumour load was scored after 3 weeks. RESULTS Sepracoat had a small but significant inhibitory effect on the adhesion of CC-531 cells in vitro. However, we were unable to repeat this effect in either rat experiment. CONCLUSION Sepracoat may inhibit adhesion of tumour cells to the mesothelium but it had no appreciable effect on intra-abdominal tumour growth in this dose in either experiment in rats.

Functional Aspects of Small-Bowel Transplantation in Rats

Although clinical small-bowel transplantation is still severely hindered by rejection of the graft, prolonged graft survival can be achieved by using cyclosporin A in several experimental models of small-bowel transplantation. In an immunologically quiescent phase after transplantation, the important question arises whether a small-bowel allograft has enough functional capacity to maintain a normal nutritional status. We investigated the functional capacity of orthotopic small-bowel transplants and evaluated the ability of the total small-bowel transplant to absorb orally given cyclosporin in the early postoperative period and the effect of this oral cyclosporin treatment on allograft survival as compared with intramuscular administration. Between 3 and 7 months postoperatively, recipients of syngeneic and allogeneic total small-bowel transplants and syngeneic jejunal segmental grafts had significantly decreased serum triglyceride levels. Total serum protein and albumin concentrations, serum cholesterol values, fecal fat excretion, and percentage of split fatty acids were normal. One year after transplantation the weight in the groups transplanted with a total small-bowel graft was not different from age-matched untreated controls. Animals grafted with a segmental graft, however, showed a significantly impaired growth and had not reached a normal weight 1 year after transplantation. Growth was also significantly impaired after near-total small-bowel resection. These animals had to be killed because of their poor condition. Cyclosporin absorption after small-bowel transplantation equalled that in normal controls. Graft survival after intramuscular treatment, however, was significantly better than after oral treatment.