Name: indocyanine green-guided intraoperative imaging to facilitate video-assisted retroperitoneal debridement for treating acute necrotizing pancreatitis
Uploaded: 2022-09-08T15:00:00
Description: Watch this Scientific Journal Video about Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis at JoVE.c

The authors thank Prof. Yu Guo and Prof. Yunpeng Hua (Department of Liver Surgery, the First Affiliated Hospital, Sun Yat-sen University) for providing advice and careful review. This work was supported by the National Natural Science Foundation of China (81201919), the Natural Science Foundation of Guangdong Province (2017A030313495).

Study protocol was approved by the ethics committee of the First Affiliated Hospital of Sun Yat-sen University and the study was conducted in accordance with the Helsinki Declaration. Written informed consents were obtained from patients.
1. Inclusion-exclusion criteria
<ol>
	<li>Include adult acute pancreatitis patients with evidence of infected necrosis that required an indication for invasive intervention. 
	NOTE: Acute pancreatitis is defined when at least two of th...

<ol>
	<li>Trikudanathan, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+concepts+in+severe+acute+and+necrotizing+pancreatitis:+An+evidence-based+approach.">Current concepts in severe acute and necrotizing pancreatitis: An evidence-based approach.</a> Gastroenterology. 156 (7), 1994-2007 (2019).</li><li>Baron, T. H., DiMaio, C. J., Wang, A. Y., Morgan, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=American+Gastroenterological+Association+clinical+practice+update:+Management+of+pancreatic+necrosis.">American Gastroenterological Association clinical practice update: Management of pancreatic necrosis.</a> Gastroenterology. 158 (1), 67-75 (2020).</li><li>Raraty, M. G. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimal+access+retroperitoneal+pancreatic+necrosectomy:+improvement+in+morbidity+and+mortality+with+a+less+invasive+approach.">Minimal access retroperitoneal pancreatic necrosectomy: improvement in morbidity and mortality with a less invasive approach.</a> Annals of Surgery. 251 (5), 787-793 (2010).</li><li>Dhingra, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+or+multiport+percutaneous+endoscopic+necrosectomy+performed+with+the+patient+under+conscious+sedation+is+a+safe+and+effective+treatment+for+infected+pancreatic+necrosis+(with+video).">Single or multiport percutaneous endoscopic necrosectomy performed with the patient under conscious sedation is a safe and effective treatment for infected pancreatic necrosis (with video).</a> Gastrointestinal Endoscopy. 81 (2), 351-359 (2015).</li><li>Gomatos, I. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Outcomes+from+minimal+access+retroperitoneal+and+open+pancreatic+necrosectomy+in+394+patients+with+necrotizing+pancreatitis.">Outcomes from minimal access retroperitoneal and open pancreatic necrosectomy in 394 patients with necrotizing pancreatitis.</a> Annals of Surgery. 263 (5), 992-1001 (2016).</li><li>Boni, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+applications+of+indocyanine+green+(ICG)+enhanced+fluorescence+in+laparoscopic+surgery.">Clinical applications of indocyanine green (ICG) enhanced fluorescence in laparoscopic surgery.</a> Surgical Endoscopy. 29 (7), 2046-2055 (2015).</li><li>Nardi, P. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+angiography+with+indocyanine+green+to+assess+anastomosis+perfusion+in+patients+undergoing+laparoscopic+colorectal+resection:+results+of+a+multicenter+randomized+controlled+trial.">Intraoperative angiography with indocyanine green to assess anastomosis perfusion in patients undergoing laparoscopic colorectal resection: results of a multicenter randomized controlled trial.</a> Surgical Endoscopy. 34 (1), 53-60 (2020).</li><li>Newton, A. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraoperative+near-infrared+imaging+can+identify+neoplasms+and+aid+in+real-time+margin+assessment+during+pancreatic+resection.">Intraoperative near-infrared imaging can identify neoplasms and aid in real-time margin assessment during pancreatic resection.</a> Annals of Surgery. 270 (1), 12-20 (2019).</li><li>Cai, Y., Zheng, Z., Gao, P., Li, Y., Peng, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+duodenum-preserving+total+pancreatic+head+resection+using+real-time+indocyanine+green+fluorescence+imaging.">Laparoscopic duodenum-preserving total pancreatic head resection using real-time indocyanine green fluorescence imaging.</a> Surgical Endoscopy. 35 (3), 1355-1361 (2021).</li><li>Santvoort, H. C. v., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dutch+pancreatitis+study+group.+A+step-up+approach+or+open+necrosectomy+for+necrotizing+pancreatitis.">Dutch pancreatitis study group. A step-up approach or open necrosectomy for necrotizing pancreatitis.</a> The New England Journal of Medicine. 362 (16), 1491-1502 (2010).</li><li>Bakker, O. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dutch+Pancreatitis+Study+Group.+Endoscopic+transgastric+vs+surgical+necrosectomy+for+infected+necrotizing+pancreatitis:+a+randomized+trial.">Dutch Pancreatitis Study Group. Endoscopic transgastric vs surgical necrosectomy for infected necrotizing pancreatitis: a randomized trial.</a> JAMA. 307 (10), 1053-1061 (2012).</li><li>Brunschot, S. v., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dutch+Pancreatitis+Study+Group.+Endoscopic+or+surgical+step-up+approach+for+infected+necrotising+pancreatitis:+a+multicentre+randomised+trial.">Dutch Pancreatitis Study Group. Endoscopic or surgical step-up approach for infected necrotising pancreatitis: a multicentre randomised trial.</a> Lancet. 391 (10115), 51-58 (2018).</li><li>Bang, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+endoscopic+transluminal+approach,+compared+with+minimally+invasive+surgery,+reduces+complications+and+costs+for+patients+with+necrotizing+pancreatitis.">An endoscopic transluminal approach, compared with minimally invasive surgery, reduces complications and costs for patients with necrotizing pancreatitis.</a> Gastroenterology. 156 (4), 1027-1040 (2019).</li></ol>

The present study reveals that ICG-guided real-time intraoperative near-infrared fluorescence imaging may provide benefit to perfusion assessment and visualization of adjacent normal tissues during debridement in VARD.
In the minimally invasive era, the step-up approach consisting of PCD or endoscopic transmural drainage followed by endoscopic necrosectomy or surgical debridement, such as VARD, has been regarded as standard treatment of severe acute necrotizing pancreatitis patients<sup class=...

Acute pancreatitis (AP) is one of the most common digestive diseases and brings enormous medical and economic burden to patients. About 20% of AP patients develop severe acute pancreatitis (SAP) that gets complicated with infected necrosis or persistent organ dysfunction1. SAP is usually associated with higher morbidity rate and mortality rate (up to 30%)1. In SAP patients with infected necrosis having persistent organ dysfunction or failure to recover after percutaneous drainage (PCD), or suffering from gastrointestinal or biliary obstruction, operative debridement should be considered1,2.
In the minimally invasive era, there are multiple approaches to operative debridement beside open surgery, including endoscopic transluminal necrosectomy, laparoscopic or open transgastric debridement, and video-assisted retroperitoneal debridement (VARD), which is the part of the step-up approach1,2. VARD is the preferred approach for patients with left-sided distribution of infected necrosis extended to paracolic gutter or deep to the retroperitoneum2. As there are rarely any visually clear separating surface in white light image under laparoscopy between necrotic debris and adjacent inflammatory normal tissues due to extensive retroperitoneal adhesions, VARD is inevitably accompanied by the risk of vascular injury, external pancreatico-cutaneous, or enterocutaneous fistulae3,4,5.
Real-time intraoperative near-infrared fluorescence imaging with indocyanine green (ICG) has been applied to facilitate perfusion assessment of bowel6,7 and visualization of biliary and vascular anatomy8,9. In view of the above disadvantages of VARD, we apply real-time near-infrared fluorescence imaging with ICG during VARD that enables the visualization of the well-perfused adjacent normal tissues and vascular structure. This modified technique (ICG-guided VARD) can provide a clear separating surface during operative debridement and reduced risk of vascular or enteric injury. ICG-guided VARD may facilitate surgeons to perform safer debridement in treating severe acute necrotizing pancreatitis.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>The 4K Ultra HD Fluorescence Endoscopic Navigation System</td>
			<td>Guangdong OptoMedic Technologies Inc</td>
			<td>OPTO-CAM214K</td>
			<td>fluorescence laparoscopy</td>
		</tr>
		<tr>
			<td>indocyanine green</td>
			<td>DanDong YiChuang Pharmaceutical CO., LTD</td>
			<td>H20055881</td>
			<td>indocyanine green injection for fluorescence imaging</td>
		</tr>
	</tbody>
</table>

indocyanine green-guided intraoperative imaging to facilitate video-assisted retroperitoneal debridement for treating acute necrotizing pancreatitis

Video-assisted retroperitoneal debridement (VARD) is a feasible, minimally invasive necrosectomy method for treating severe acute necrotizing pancreatitis, if it does not resolve or is accompanied with infected necrosis in the retroperitoneum. As there are rarely any visually clear separating surface in white light image between necrotic debris and adjacent inflammatory normal tissues due to extensive retroperitoneal adhesions, VARD is accompanied with the risk of vascular injury, external pancreatico-cutaneous or enterocutaneous fistulae. In view of the above disadvantages, we apply real-time intraoperative near-infrared fluorescence imaging with indocyanine green (ICG) during VARD, which enables visualization of the well-perfused adjacent normal tissues. This modified technique (ICG-guided VARD) can provide a clear separating surface during debridement and reduce the risk of vascular or enteric injury. ICG-guided VARD may facilitate surgeons to perform safer debridement in treating severe acute necrotizing pancreatitis.

ICG-guided VARD had been successfully performed in three severe acute necrotizing pancreatitis patients from June 2021. Characteristics of these patients at baseline and after VARD are included in Table 1. The first patient who received ICG-guided VARD was a male, 41-year-old patient who was admitted on 20th June 2021. He suffered from moderately acute necrotizing pancreatitis. Abdominal contrast enhanced CT scan revealed (as shown in Figure 1 and Table 1) t...

Watch this Scientific Journal Video about Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis at JoVE.c

Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis

This protocol presents Indocyanine Green-guided video-assisted retroperitoneal debridement (ICG-guided VARD) for treating severe acute necrotizing pancreatitis.

Video-assisted retroperitoneal debridement (VARD) is a feasible, minimally invasive necrosectomy method for treating severe acute necrotizing ...

This protocol presents indocyanine green-guided video-assisted retroperitoneal debridement for treating severe acute necrotizing pancreatitis. The ICG-guided VARD is an easy and feasible approach to visualize the well perfused adjacent normal tissues and vascular structure of mesenteric vessels. Begin by inserting a 12 millimeter laparoscopic trocar along the dilated tract to the retroperitoneal necrotic collections.Apply carbon dioxide pneumoretroperitoneum if a wider debridement space is needed by insufflating carbon dioxide through the 12 millimeter trocar. Then place a near infrared fluorescence laparoscopic via the observing trocar and make an additional 10 millimeter incision subcostal in the left or right flank at the mid-axillary line as described in the text manuscript. Insert the 10 millimeter operative trocar and switch the display mode of the laparoscopic to multi-display mode Before separating necrotic debris from adjacent normal tissue.Inject the first bolus of indocyanine green or ICG intravenously in a peripheral vein, then flush with 10 milliliters of saline. After 10 to 20 seconds, visualize peak perfusion of adjacent normal tissues or vessels in the fluorescence field. A more distinct separating surface will be distinguishable from the debris.Remove the poorly perfused and loosely adherent necrotic debris using laparoscopic graspers. Avoid tearing the underlying bowel or vessel. Inject another bolus of ICG intravenously if ICG fluorescence decay.After removing the bulk of necrotic debris, irrigate the cavity of the retroperitoneum with saline and aspirate with a laparoscopic aspirator until the lavage fluid becomes clear. Place at least one pair of drainage tubes in the deepest region of the cavity after debridement before proceeding with the fascia and skin suturing. The 41-year-old male patient suffered from moderately acute necrotizing pancreatitis and received the ICG-guided VARD.Abdominal contrast enhanced CT scan revealed that the necrotic collections consist of the lesser arc peripancreatic space and extended to the bilateral retroperitoneum. Infected necrosis in the right retroperitoneum was persistent after initial percutaneous drainage and the first VARD was performed about four weeks after the onset of pancreatitis. The ICG perfusion of adjacent normal tissues or vessels was visualized in the fluorescence field and a clearer separating surface was distinguished from the debris.Infected necrosis was resolved and drainage tubes were removed seven days after ICG-guided VARD. ICG-guided VARD warrants further randomized clinical trials to confirm its practicability and safely treating acute necrotizing pancreatitis.

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Medicine

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration have been developed. The ideal animal model should closely mimic the human intervertebral disc with regard to morphology, biomechanical properties and the absence of notochordal cells. The sheep lumbar intervertebral disc model fulfils these criteria. We present an ovine model of intervertebral disc degeneration utilizing a drill bit injury through a lateral retroperitoneal approach. The lateral approach significantly reduces the incision and potential morbidity associated with the traditional anterior approach to the ovine spine. Utilization of a drill-bit method of injury affords the ability to produce a consistent and reproducible injury, of precise dimensions, that initiates a consistent degree of intervertebral disc degeneration. The focal nature of the annular and nucleus pulposus defect more closely mimics the clinical condition of focal intervertebral disc herniation. Sheep recover rapidly following this procedure and are typically mobile and eating within the hour. Intervertebral disc degeneration ensues and sheep undergo necropsy and subsequent analysis at periods from eight weeks. We believe that the drill bit injury model of intervertebral disc degeneration offers advantages over more conventional annular injury models.

Intervertebral disc degeneration is a significant contributor to back pain and a leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration exist. We demonstrate an ovine model of intervertebral disc degeneration, utilizing a drill bit, which achieves a consistent disc injury and reproducible level of disc degeneration.

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous ...

Robotic Lateral Pancreaticojejunostomy for Chronic Pancreatitis

Lateral pancreaticojejunostomy (LPJ) has shown good postoperative outcomes in patients with painful, morphine dependent, chronic pancreatitis (CP). The recent rise of robotic and laparoscopic pancreatic surgery has found benefits such as reduced time to functional recovery. Few studies have reported on the feasibility, technique and outcome of robotic LPJ, especially including transection of the gastroduodenal artery. The present study describes the main steps for robotic LPJ in a patient with painful chronic pancreatitis with a dilated main pancreatic duct. The patient underwent robotic LPJ. The LPJ anastomosis is performed using a running suture technique in a longitudinal side-to-side manner. Routinely, the gastroduodenal artery is transected to drain the entire length of the main pancreatic duct. The patient is in French position; 7 trocars are placed (4 robotic, 2 laparoscopic assistants, 1 liver-retractor). After docking of the robot system, the omental bursa is opened, and the right gastroepiploic artery and vein are ligated at their base at the lower border of the pancreas. Intraoperative ultrasonography is performed in order to determine trajectory of the dilated main pancreatic duct which is opened for its entire length after the gastroduodenal artery has been suture ligated both cranially and caudally from the main pancreatic duct. A Roux limb is created, and a latero-lateral PJ is fashioned using several 3-0 barbed sutures. A stapled jejuno-jejunostomy is created at sufficient distance from the pancreatic anastomosis, aided by a 50 cm suture. The described technique for robotic LPJ is a complex but feasible operation for patients with treatment refractory CP and a dilated main pancreatic duct. Due to its complexity, implementation in high volume centers with extensive experience with CP surgery may improve outcomes.

Robotic lateral pancreaticojejunostomy (RLPJ) may be used in patients with painful, morphine dependent, chronic pancreatitis and a dilated main pancreatic duct. We describe a standardized and reproducible technique for RLPJ, which includes transection of the gastroduodenal artery.

Lateral pancreaticojejunostomy (LPJ) has shown good postoperative outcomes in patients with painful, morphine dependent, chronic pancreatitis (CP). The ...

Posterior Approach for Debridement of the Psoas Abscess

This method focuses on outlining a safe zone for irrigation and debridement of a psoas abscess through a posterior approach. Initially, an anterior approach to the spine was performed to ensure that the anterior longitudinal ligament and the psoas muscle could be visualized. All the abdominal organs were removed. Subsequently, a posterior approach was performed to remove the paraspinal muscles from L1&#8211;L5. The transverse processes, pars interarticularis and lamina of L1&#8211;L5 were identified. The exiting nerve root was identified between the transverse processes and followed into the substance of the psoas muscle. Using the anterior and posterior approach, the lumbar plexus was isolated from the substance of the psoas muscle. Before and after various steps of dissection, digital photographs were obtained. These images were uploaded into ImageJ and multiple measurements, including the distance between the lateral superior and inferior tip of each TP to the most lateral region of the plexus, the distance between the lateral superior and inferior tip of the TP to the lateral edge of the psoas, and the width of the lumbar plexus were recorded. The safe zone for entering the substance of the psoas muscle was defined between the lateral edge of the psoas muscle and the lateral edge of the lumbar plexus. The relationship of this interval to the tip of the transverse process at each level was measured and reported.

This is a cadaveric study investigating the landmarks for the posterior approach for irrigation and debridement of the psoas abscess. The interval between the transverse processes (TP) was used to access the substance of the psoas muscle.

This method focuses on outlining a safe zone for irrigation and debridement of a psoas abscess through a posterior approach. Initially, an anterior ...

Sodium Taurocholate Induced Severe Acute Pancreatitis in C57BL/6 Mice

Biliary acute pancreatitis induction by sodium taurocholate infusion has been widely used by the scientific community due to the representation of the human clinical condition and reproduction of inflammatory events corresponding to the onset of clinical biliary pancreatitis. The severity of pancreatic damage can be assessed by measuring the concentration, speed, and volume of the infused bile acid. This study provides an updated checklist of the materials and methods used in the protocol reproduction and shows the main results from this acute pancreatitis (AP) model. Most of the previous publications have limited themselves to reproducing this model in rats. We have applied this method in mice, which provides additional advantages (i.e., the availability of an arsenal of reagents and antibodies for these animals along with the possibility of working with genetically modified strains of mice) that may be relevant to the study. For acute pancreatitis induction in mice, we present a systematic protocol, with a defined dose of 2.5% sodium taurocholate at an infusion speed 10 &#181;L/min for 3 min in C57BL/6 mice that reaches its maximal level of severity within 12 h of induction and highlight results with outcomes that validate the method. With practice and technique, the total estimated time, from the induction of anesthesia to the completion of the infusion, is 25 min per animal.

Animal models for severe acute pancreatitis enable the study of pathophysiological changes at the initial stage, facilitating observation of the evolution of inflammatory events. Here we provide a protocol for the induction of severe acute biliary pancreatitis by retrograde infusion of sodium taurocholate into the pancreatic duct of anesthetized C57BL/6 mice.

Biliary acute pancreatitis induction by sodium taurocholate infusion has been widely used by the scientific community due to the representation of the ...

Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a lack of effective treatments in the current clinical practice. With the easy accessibility of transgenic and knockout strains and their small size, which allows minimal doses of drugs required for in vivo evaluation, a well-established experimental model in mice is preferred for AP research. Moreover, SAP induced through sodium taurocholate (TC) is currently one of the most widely used and best characterized models. This model has been investigated for novel therapies and possible molecular events during the process of AP. Here, we present the generation of an AP mouse model using sodium taurocholate and a simple homemade microsyringe. Moreover, we also provide the methodology for the subsequent histology and serological testing.

A mouse model of severe acute pancreatitis is described herein. The procedure presented here is very rapid, simple, and accessible, thereby potentially allowing the study of the molecular mechanisms and different therapeutic interventions in acute pancreatitis in a convenient way.

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a ...

Robotic Left Hepatectomy using Indocyanine Green Fluorescence Imaging for an Intrahepatic Complex Biliary Cyst

Biliary cysts (BC) are rare congenital dilatations of intra- and extrahepatic parts of the biliary tract and bear a significant risk of carcinogenesis. Surgery is the cornerstone treatment for patients with BC. While total BC excision and Roux-Y hepaticojejunostomy is the treatment method of the choice in patients with extrahepatic BC (i.e., Todani I-IV), patients with intrahepatic BC (i.e., Todani V) benefit the most from a surgical liver resection. In recent years, minimally invasive liver surgery (MILS) including robotic MILS has gained more acceptance as a feasible, safe, and effective procedure for the treatment of both benign and malignant indications. Robotic major MILS is still considered technically demanding and a detailed description of the technical approach during robotic major MILS has only been limitedly discussed in the literature. The current article describes the main steps for a robotic left hepatectomy in a patient with a large BC Todani Type V. The patient is in French position with 5 trocars placed (4 robotic, 1 laparoscopic assistant). After mobilizing the left hemiliver, the left and right hepatic artery are dissected carefully followed by a cholecystectomy. Intraoperative ultrasound is performed to confirm localization and margins of the BC. The Left hepatic artery and left portal vein are isolated, clipped, and divided. Indocyanine green (ICG) fluorescence imaging is used regularly during the entire procedure to visualize and confirm biliary tract anatomy and the BC. Parenchymal transection is performed with robotic cautery hook for the superficial part and robotic cautery spatula, bipolar cautery, and vessel sealer for the deeper parenchyma. The postoperative course was uncomplicated. A robotic left hepatectomy is technically demanding, yet a feasible and safe procedure. ICG-fluorescence imaging aids in delineating the BC and bile duct anatomy. Further, comparative studies are needed to confirm clinical benefits of robotic MILS for benign and malignant indications.

Robotic liver surgery has gained more acceptance as a feasible, safe, and effective procedure for the treatment of both benign and malignant indications. However, robotic left hepatectomy is still technically demanding. We describe our surgical technique of a robotic left hepatectomy using indocyanine green fluorescence imaging for a large biliary cyst.

Biliary cysts (BC) are rare congenital dilatations of intra- and extrahepatic parts of the biliary tract and bear a significant risk of carcinogenesis. ...

In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis

Necrotizing enterocolitis (NEC) is a devastating disease affecting preterm infants, characterized by intestinal inflammation and necrosis. Enteroids have recently emerged as a promising system to model gastrointestinal pathologies. However, currently utilized methods for enteroid manipulation either lack access to the apical surface of the epithelium (three-dimensional [3D]) or are time-consuming and resource-intensive (two-dimensional [2D] monolayers). These methods often require additional steps, such as microinjection, for the model to become physiologically translatable. Here, we describe a physiologically relevant and inexpensive protocol for studying NEC in vitro by reversing enteroid polarity, resulting in the apical surface facing outward (apical-out). An immunofluorescent staining protocol to examine enteroid barrier integrity and junctional protein expression following exposure to tumor necrosis factor-alpha (TNF-&#945;) or lipopolysaccharide (LPS) under normoxic or hypoxic conditions is also provided. The viability of 3D apical-out enteroids exposed to normoxic or hypoxic LPS or TNF-&#945; for 24 h is also evaluated. Enteroids exposed to either LPS or TNF-&#945;, in combination with hypoxia, exhibited disruption of epithelial architecture, a loss of adherens junction protein expression, and a reduction in cell viability. This protocol describes a new apical-out NEC-in-a-dish model which presents a physiologically relevant and cost-effective platform to identify potential epithelial targets for NEC therapies and study the preterm intestinal response to therapeutics.

In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis

This protocol describes an apical-out necrotizing enterocolitis (NEC)-in-a-dish model utilizing small intestinal enteroids with reversed polarity, allowing access to the apical surface. We provide an immunofluorescent staining protocol to detect NEC-related epithelial disruption and a method to determine the viability of apical-out enteroids subjected to the NEC-in-a-dish protocol.

Necrotizing enterocolitis (NEC) is a devastating disease affecting preterm infants, characterized by intestinal inflammation and necrosis. Enteroids have ...

Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is similar in color to the thyroid gland. The lack of effective animal models in preclinical research is a severe limitation for the development of PG identification techniques. This protocol allows for the establishment of a simple and effective rat model for PG identification. In this model, black iron oxide nanoparticles (IONPs) are injected locally in the thyroid gland and rapidly diffuse within the thyroid gland but not the PG. A negatively stained PG and a positively stained thyroid gland can be easily identified by the naked eye without requiring external microscopes. The position of the PG can be identified by increasing the color contrast between the thyroid gland and the PG, based on the color of the black IONPs. This rat model is low-cost and convenient for PG identification, and the IONPs are a novel PG contrast agent.

To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent.

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is ...