Hyperbaric oxygen improves capillary morphology in severe acute pancreatitis.

OBJECTIVES: This article aims to determine the effect of acute pancreatitis on microvascular morphology and the impact of treatment with hyperbaric oxygen (HBO). METHODS: Sixty-seven male Wistar rats were induced with acute pancreatitis by retrograde bile duct injection. Rats were randomized to 12-hourly HBO or control treatment. Two rats in each group were killed at baseline and 24, 48, and 72 hours postinduction, and a cast of the pancreatic microvasculature was examined using scanning electron microscopy. RESULTS: Normal pancreatic vasculature is a dense network with a consistent capillary diameter. In acute pancreatitis, mean capillary diameter is increased at 24 hours (P < 0.001) and further increased at 48 hours (P = 0.007). From 24 hours, diameter heterogeneity is increased (P < 0.001) and capillary density is reduced (P < 0.001). Hyperbaric oxygen has a significant effect on vascular morphology changes from 48 hours after induction. Capillary diameter and heterogeneity of diameter are decreased by HBO (both P < 0.001). Capillary density is increased by HBO at 48 and 72 hours (P < 0.001). CONCLUSIONS: In acute pancreatitis, structural capillary diameter and heterogeneity of diameter increase and capillary density decreases. These parameters are all improved by HBO treatment. Hyperbaric oxygen treatment normalizes the pancreatic microvasculature after acute pancreatitis and may be a potentially effective treatment of this disease.

Hyperbaric oxygen therapy for severe acute pancreatitis.

Despite improvements in the supportive management of severe acute pancreatitis over the last decade, the morbidity and mortality rate remains high. The main feature of this condition is pancreatic necrosis leading to sepsis, with both localized and systemic inflammatory response syndromes. Early pathophysiological changes of the pancreas include alterations in microcirculation, ischemia reperfusion injury, and leukocyte and cytokine activation. The efficacy of hyperbaric oxygen (HBO) therapy in improving these pathophysiological disturbances is documented for various conditions. However, its effect in the treatment of severe acute pancreatitis is undetermined. This report documents the case of a 56-year-old woman presenting with severe acute pancreatitis treated by HBO therapy. The severity of disease was based on an Acute Physiology and Chronic Health Evaluation (APACHE II) illness grading score of 11 and a Baltazar based computed tomography severity index (CTSI) score of 9. Administration of 100% oxygen was commenced within 72 h of presentation at a pressure of 2.5 atmospheres for 90 min and given twice daily for a total of 5 days. Therapy was well tolerated with improvements in APACHE II and CTSI grading scores. HBO therapy for severe acute pancreatitis appeared to be safe and may have a role in improving treatment outcomes. Further study is required.

Hyperbaric oxygen therapy reduces severity and improves survival in severe acute pancreatitis.

Severe acute pancreatitis is characterized by pancreatic necrosis, resulting in local and systemic inflammation. Hyperbaric oxygen (HBO) therapy modulates inflammation, but has not been extensively studied in pancreatitis. This study investigates the effects of HBO in a rat model of severe acute pancreatitis. Sixty-four rats were induced with severe pancreatitis using 4% sodium taurocholate and randomized to HBO treatment or control. HBO was commenced 6 h after induction (100% oxygen at 2.5 atmospheres for 90 min) and continued every 12 h for a maximum of eight treatment episodes. Surviving animals were killed at 7 days. Severity of pancreatitis was graded macroscopically and microscopically. Lung edema was calculated using wet and dry lung weights. Macroscopic and microscopic severity scores (mean +/- SE) of HBO-treated animals with pancreatitis (8.3 +/- 0.7; 9.6 +/- 0.4) were lower than those of controls (10.5 +/- 0.5; 11.1 +/- 0.4) (p = 0.02 and p = 0.03, respectively). The HBO-treated group had reduced pancreatic necrosis compared to controls (40 +/- 4% vs. 54 +/- 4%; p = 0.003). There was no difference in pulmonary edema between the groups. Median survival in the HBO-treatment group was 51 h, compared to 26 h in controls. Day-7 survival was significantly improved in the HBO-treated animals compared to controls (40% vs. 27%; p = 0.04). HBO therapy reduces overall severity, decreases the extent of necrosis, and improves survival in severe acute pancreatitis.

Potential effects of hyperbaric oxygen therapy in acute pancreatitis.

BACKGROUND: To extract from the biomedical published reports, the effects of hyperbaric oxygen (HBO) on inflammatory disease, in particular acute pancreatitis. METHODS: This review will explain these effects and evaluate potential mechanisms of action of HBO in acute pancreatitis. A Medline/PubMed search (January 1966 to July 2004) with manual cross-referencing was conducted, including all relevant articles investigating the molecular and systemic effects of HBO on inflammatory diseases, particularly focusing on the studies of acute pancreatitis. All publication types, languages and subsets were searched. RESULTS: Original and review articles and short communications were extracted. The selected original articles covered the molecular and systemic effects of HBO and the effects in inflammatory disease states. The major findings are that HBO can act as an anti-inflammatory agent and as an antimicrobial agent. Many of the effects of HBO would be beneficial in the treatment of acute severe pancreatitis. Work carried out to date in animal models of acute pancreatitis shows promising improvements in severity but studies are limited to date. CONCLUSION: Acute pancreatitis impairs the pancreatic and systemic microcirculation and causes acute inflammation. These processes are potentially improved by HBO therapy.

Altered growth patterns of colorectal liver metastases after thermal ablation.

BACKGROUND: Thermal ablation by radiofrequency or laser is used increasingly for the treatment of colorectal liver metastases. Recurrence after thermal ablation is common and occurs both locally and at distant sites. One possible cause of this recurrence may be a result of growth stimulation of micrometastases in the remaining liver. This study examined the impact of thermal ablation on growth patterns of hepatic micrometastases. METHODS: Colorectal liver metastases were induced in male CBA-strain mice via an intrasplenic injection of a murine-derived cancer cell line. Subtotal thermal ablation of the left posterior lobe of the liver (30% of total liver volume) was performed by neodymium yttrium-aluminum-garnet laser 7 days after induction of metastases. The distribution, number, cross-sectional diameter, volume, and proliferation rate of established neoplasms were compared with controls at 21 days after tumor induction. The effect of thermal ablation of 7% of the total liver volume by laser on the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor 2 (FGF-2), transforming growth factor beta, and cellular proliferation (Ki-67 antigen) adjacent to the ablated site was assessed by immunohistochemistry in separate groups of animals at specific time points after therapy. RESULTS: Thermal ablation did not alter the overall volume, number, size, and proliferation rate of neoplasms 21 days after laser ablation. There were no extrahepatic metastases after therapy. The number of neoplasms in the regenerated posterior lobe was equivalent to control despite subtotal ablation (29 +/- 2 vs 27 +/- 2; P = NS). A greater amount of metastases occupied the regenerated thermal-ablated lobe compared with controls (55% +/- 4% vs 29% +/- 3%; P < .04). Thermal ablation stimulated liver proliferation adjacent to the treatment site at 12 hours compared with untreated controls. Stimulation peaked at 72 hours (20% +/- 1% vs 1% +/- 1%; P < .001) and persisted to 21 days after therapy. FGF-2 and VEGF expression increased in liver tissue adjacent to the ablation site compared with baseline, peaking at 12 hours (112% +/- 2% vs 102% +/- 1%; P < .001) and 72 hours (114% +/- 2% vs 101% +/- 1%; P < .001), respectively. CONCLUSIONS: Thermal ablation promotes the progression of micrometastases to form macroscopically detectable neoplasms in treated regenerating liver. This effect may relate to an increased expression of VEGF and FGF-2 adjacent to the treatment site.

Mechanisms of focal heat destruction of liver tumors.

BACKGROUND: Focal heat destruction has emerged as an effective treatment strategy in selected patients with malignant liver tumors. Radiofrequency ablation, interstitial laser thermotherapy, and microwave treatment are currently the most widely applied thermal ablative techniques. A major limitation of these therapies is incomplete tumor destruction and overall high recurrences. An understanding of the mechanisms of tissue injury induced by focal hyperthermia is essential to ensure more complete tumor destruction. Here, the currently available scientific literature concerning the underlying mechanisms involved in the destruction of liver tumors by focal hyperthermia is reviewed. METHODS: Medline was searched from 1960 to 2004 for literature regarding the use of focal hyperthermia for the treatment of liver tumors. All relevant literature was searched for further references. RESULTS: Experimental evidence suggests that focal hyperthermic injury occurs in two distinct phases. The first phase results in direct heat injury that is determined by the total thermal energy applied, tumor biology, and the tumor microenvironment. Tumors are more susceptible to heat injury than normal cells as the result of specific biological features, reduced heat dissipating ability, and lower interstitial pH. The second phase of hyperthermic injury is indirect tissue damage that produces a progression of tissue injury after the cessation of the initial heat stimulus. This progressive injury may involve a balance of several factors, including apoptosis, microvascular damage, ischemia-reperfusion injury, Kupffer cell activation, altered cytokine expression, and alterations in the immune response. Blood flow modulation and administration of thermosensitizing agents are two methods currently used to increase the extent of direct thermal injury. The processes involved in the progression of thermal injury and therapies that may potentially modulate them remain poorly understood. CONCLUSION: Focal hyperthermia for the treatment of liver tumors involves complex mechanisms. Evidence suggests that focal hyperthermia produces both direct and indirect tissue injury by differing underlying processes. Methods to enhance the effects of treatment to achieve complete tumor destruction should focus on manipulating these processes.

Progressive microvascular injury in liver and colorectal liver metastases following laser induced focal hyperthermia therapy.

BACKGROUND AND OBJECTIVES: Focal hyperthermia by laser or radiofrequency is currently the preferred method for local ablation of liver tumors. The underlying mechanism of action of focal hyperthermia, in particular the relationship between the microvascular and tissue effect is uncertain and was investigated in a murine model. STUDY DESIGN/MATERIALS AND METHODS: Focal hyperthermia produced by a Neodymium-Yttrium-Aluminium-Garnet laser (wavelength 1,064 nm) was applied to the normal liver and colorectal cancer liver metastases in inbred male CBA strain mice at 2 W for 50 seconds (100 J). Tissue injury was assessed at 0, 24, 48, 72, 120, and 168 hours following therapy by measurements of necrosis in tissue sections stained for nicotinamide adenine dinucleotide (NADH) diaphorase activity. Characteristics of microvascular injury were assessed at the various time points by scanning electron microscopy (SEM) of vascular resin casts, Laser Doppler flowmetry, and confocal in vivo microscopy. RESULTS: Focal hyperthermia produced progressive tissue and vascular injury. There was an initial reduction in blood flow and increased vascular permeability in the microcirculation of both tumor and liver tissue immediately following heat application as assessed by laser Doppler flowmetry and confocal in vivo microscopy, respectively. SEM of vascular casts showed heterogeneous regions of microvascular injury immediately following heat application. The extent of initial vascular disruption or occlusion on SEM of vascular resin casts (mean+/-SE) was significantly less than the tissue necrosis in liver (1.9+/-0.1 mm vs. 3.0 mm+/-0.2 mm P<0.001), but was equivalent to the tissue injury in tumor tissue (3.5 mm+/-0.2 mm vs. 3.6 mm+/-0.1 mm P = 0.907). This was followed by a progressive increase in both microvascular and tissue injury in liver and tumor that peaked by 72 hours following treatment. The peak microvascular injury and tissue damage in liver (6.6 mm+/-0.2 and 6.3 mm+/-0.2 mm, respectively) was significantly greater than the extent of microvascular and tissue damage in tumors (4.8 mm+/-0.2 mm and 4.5 mm+/-0.2 mm, respectively) (P<0.001). The progression of microvascular injury in liver and tumor at times preceded the tissue injury. CONCLUSION: Focal hyperthermia produces both progressive microvascular and tissue damage in liver and colorectal liver metastases. An increase in tissue injury following focal hyperthermia may be a direct result of progressive microvascular damage. Tumor vessels appear more susceptible to direct focal hyperthermia destruction than liver sinusoids. The liver sinusoids are however more susceptible to progressive damage or occlusion following the initial laser thermal stimulus.

Comparison of 980- and 1064-nm wavelengths for interstitial laser thermotherapy of the liver.

OBJECTIVE: Interstitial laser thermotherapy (ILT) of liver tumors is generally performed using neodyium yttrium-aluminium-garnet (Nd-YAG) lasers. More versatile diode units, developed predominantly for other clinical applications, may be equally suitable for ILT. This study compares the efficacy of diode and Nd-YAG lasers in achieving maximum tissue necrosis, at low power, in a murine model. METHODS: Thermal ablation was induced in the liver of CBA strain mice by diode (980-nm wavelength) and Nd-YAG (1064-nm wavelength) lasers using 400-microm diameter bare fibers. Treatment time prior to tissue charring was determined for both lasers at a power output of 2 W. Tissue temperature was recorded upon completion of therapy 3 mm from the fiber insertion site. The maximum diameter of necrosis was accurately assessed by nicotinamide adenine dinucleotide (NADH) diaphorase tissue staining. RESULTS: Maximum diameter of tissue necrosis prior to charring occurred at 20 s (40 J) with the diode laser compared to 50 s (100 J) with the Nd-YAG laser. The maximum diameter of necrosis (mean [SEM]) produced by the diode laser, 5.9 mm (0.14), was equivalent to the necrosis induced by the Nd-YAG laser, 5.9 mm (0.14) (p = 0.963). Tissue temperature 3 mm from the fiber application site immediately following ILT in the diode laser group, 40.8 degrees C (0.93), was not statistically different than that of the Nd-YAG laser group, 39.0 degrees C (0.86) (p = 0.452). Tissue charring consistently prevented treatment beyond 20 s at 2W by the diode laser. CONCLUSION: Low power ILT with diode and Nd-YAG lasers achieves equivalent maximal necrosis when applied to the liver by a bare fiber. Treatment time to produce maximal necrosis is however significantly shorter with the diode laser.

The apoptotic response of liver and colorectal liver metastases to focal hyperthermic injury.

BACKGROUND: Ablation of liver tumours by focal hyperthermia causes tissue injury not only by direct effects, but also by progressive tissue damage following the initial heat application. The mechanism by which this progressive injury occurs remains undefined. Stimulation of apoptosis following the initial heat stimulus may be involved in this progression. The role of apoptosis in the subsequent progression of focal hyperthermia injury was investigated in liver and colorectal liver metastases in a murine model. MATERIALS AND METHODS: Focal hyperthermia produced by laser (Nd-YAG--wavelength 1064 nm) was applied to the liver and colorectal liver metastases in CBA mice (2 Watts for 50 seconds). The animals were killed at 0, 12, 24, 48, 72, 120 and 168 hours after the application of focal hyperthermia. Haematoxylin and eosin staining and immunohistochemistry were performed on paraffin sections to assess the extent of tissue necrosis. Apoptosis was examined by assessment of DNA fragmentation using terminal deoxynucleotidyl transferase d-uridine triphosphate nick end labelling (TUNEL) and activated Caspase 3 immunostaining. The sequence of the apoptotic response was determined at the treated tissue margins and compared to untreated liver and tumour. RESULTS: Focal hyperthermia produced progressive tissue injury in both liver and colorectal liver metastases. TUNEL labelling was less specific than activated Caspase 3 in detecting apoptotic cells. Apoptosis was detected and peaked at 12 hours following therapy based on activated Caspase 3 staining, before returning to baseline at 72 hours. In tumour tissue, the apoptotic response was more sustained, peaking at 24 hours before returning to baseline levels by 96 hours following therapy. CONCLUSION: Increased apoptosis occurs following the application of focal hyperthermia to normal liver and colorectal metastases. The apoptotic response is more sustained in tumour tissue and contributes to the progressive injury that is evident after the initial heat stimulus.

Focal hyperthermia produces progressive tumor necrosis independent of the initial thermal effects.

Focal hyperthermia, produced using laser, radio frequency, and microwave, is used to treat liver tumors. The exact mechanisms of tissue destruction using focal hyperthermia are, however, unknown. Clinical and experimental studies suggest a progression of injury after cessation of the initial heat stimulus. This study investigates the mechanisms and time sequence of progressive tissue necrosis induced using focal hyperthermia in a murine model of colorectal liver metastases. Focal hyperthermia produced using a neodymium-yttrium aluminum garnet (Nd-YAG) laser source was applied to the normal liver and colorectal cancer liver metastases in inbred male CBA strain mice. The extent of direct lethal thermal injury was assessed histochemically using vital stain for nicotinamide adenine dinucleotide (NADH) diaphorase immediately after laser application. Tissue injury at subsequent time points was assessed using both NADH diaphorase staining and routine histology to determine the temporal relationship between tissue necrosis and time. Thermal injury occurring immediately after the application of 100 joules of energy was greater in the tumor tissue than in the normal liver (mean [standard error of the mean (SEM)]), measuring 23.5 (3.4) and 16.3 (2.6) mm(3), respectively (P=0.046), despite similar tissue temperature profiles. There was a significant increase in tissue necrosis after initial injury that was greater in the normal liver than in the tumor tissue. In the normal liver, the peak volume of necrosis was 137.4 (9.8) mm(3) and occurred at 3 days, whereas in the tumor tissue the peak was 49.0 (3.5) mm(3) at 4.5 days (P < 0.001). Focal hyperthermia produces tissue necrosis that occurs in two phases. The first phase is caused by the direct lethal thermal injury followed by a second phase involving a progression of necrosis beyond the initial thermal effects. The normal liver and the tumor tissue responded differently to focal hyperthermia. In the tumor tissue, the direct injury is more pronounced, whereas the progression of injury is more rapid and extensive in the normal liver.

Long-term survival of patients with unresectable colorectal liver metastases treated by percutaneous interstitial laser thermotherapy.

In situ ablation of colorectal cancer (CRC) liver metastases is an accepted form of treatment for selected patients. It is associated with low morbidity and mortality and increases the number of patients who may benefit from therapy compared to resection alone. This study assesses the impact of interstitial laser thermotherapy (ILT) on local tumor control and long-term survival in patients with unresectable CRC liver metastases. Percutaneous ILT was performed in patients with unresectable CRC liver metastases between January 1992 and December 1999 using a bare-tip quartz fiber connected to an Nd:YAG laser source. This was prior to the routine use of a diffusing fiber for ablative therapy. Treatment was monitored with real-time ultrasonography. Tumors were considered unresectable based on their anatomic location or the extent of liver involvement. Patients with extrahepatic disease, more than five liver metastases, or tumors larger than 10 cm in diameter were excluded from this study. Local tumor control was assessed by dynamic computed tomography (CT) 6 months after therapy. Long-term follow-up was undertaken, and the impact of various factors on survival was analyzed. Eighty patients with a mean age of 63.8 years were suitable for ILT. In total, 168 liver tumors with a median diameter of 5 cm (range 1-10 cm) were so treated. There were no procedure-related deaths. The overall complication rate was 16%, with all cases managed conservatively. Bradycardia (n = 5), pneumothorax (n = 3), and persistent pyrexia (n = 3) were the most common complications. Complete tumor ablation was noted in 67% of patients assessed by CT 6 months following the initial therapy. Median follow-up was 35 months (range 4-96 months), with 10 patients alive at the end of this period. Altogether there were 67 deaths, which were related to hepatic disease in 55 cases and to extrahepatic disease in 9; they were unrelated to malignancy in 3 others. Three patients were excluded from follow-up after ILT down-staging of tumors that allowed complete surgical resection. The median disease-free survival of patients treated by ILT was 24.6 months, with a 5-year survival of 3.8%. Poor tumor differentiation and the presence of more than two hepatic metastases were associated with lower overall survival (p < 0.01). Fourteen patients treated by ILT for postoperative hepatic recurrences had the best outcome, with a median overall survival of 36.3 months and a 5-year survival of 17.2%. Percutaneous ILT is a minimally invasive, safe, effective technique that appears to improve overall survival in specific patients with unresectable CRC liver metastases, compared to the natural history of untreated disease reported in the literature.

Effect of interstitial laser hyperthermia in a murine model of colorectal liver metastases: scanning electron microscopic study.

Interstitial laser hyperthermia (ILH) is an in situ ablative technique used to treat colorectal liver metastases. The relatively high recurrence of tumor after treatment by ILH may be related to incomplete destruction. Little is known about the effectiveness of ILH for destroying tumor microvasculature. The aim of this study was to define the changes to the microvascular architecture of tumors after treatment with ILH, specifically focusing on the completeness of tumor vasculature destruction. An intrasplenic induction model of liver metastases in 4- to 6-week-old male inbred CBA mice was used. Laser hyperthermia was applied to liver and tumor tissue using a bare optical quartz fiber from a Medilas Fibertom 4100 Nd:YAG surgical laser generator. The animals underwent microvascular corrosion casting of the livers immediately after application of ILH. Microvascular casts were then prepared and studied by scanning electron microscopy. ILH produced complete, uniform destruction of the tumor microvasculature with compete hemostasis. Blood flow in vessels larger than 100 microm diameter had a relatively protective effect, although ILH was able to overcome this barrier effectively by increasing the energy applied. ILH produces complete destruction of tumor microvasculature with hemostasis. The protective effect of blood flow in larger vessels can be overcome by the appropriate use of higher energy levels.

Interstitial laser hyperthermia and the biological characteristics of tumor: study in a murine model of colorectal liver metastases.

BACKGROUND DATA: Percutaneously applied interstitial laser hyperthermia (ILH) is a minimally invasive therapy that is currently used in the treatment of liver metastases. Despite its documented efficacy, theoretical considerations and evidence based on animal studies suggest the potential for stimulating tumor growth, especially following surgery. This study investigates the influence of ILH on tumor behaviour in an animal model of colorectal liver metastases. MATERIALS AND METHODS: A model of colorectal cancer liver metastases in male inbred CBA mice was used. Laser hyperthermia was applied to tumor tissue using a bare optical quartz from a Medilas fibertom 4100 Nd:YAG surgical laser generator. Liver injury by ILH was initially produced in three experimental groups of animals at different time points in the development of metastases. ILH was applied (i) to normal liver 10 days prior to tumor induction, (ii) immediately prior to tumor induction, and (iii) 15 days after tumor induction to achieve approximately 8% liver destruction. Animals were killed 21 days after tumor induction, and the effects of ILH on overall tumor development were compared with controls using stereological assessment of tumor volume and by histology. In a separate experimental group, the effects of ILH on fully established tumors were examined. Suitable tumors were selected 21 days after induction and partially destroyed by ILH at a standard energy setting. Animals were then killed 15 days later, and the growth rate of the residual viable tumors was compared to control tumors having undergone sham procedures. RESULTS: No significant stimulation of tumor growth was evident in any of the experimental groups following ILH, irrespective of the time of application. Incomplete tumor destruction also had no influence on subsequent tumor growth. CONCLUSION: ILH does not influence the biological characteristics of tumors during any stage of the metastatic process.