Use Of Fluorescence-guided Surgery Research Articles

Fluorescence-guided surgery in patients with tumors near motor areas of the brain

There is no common concept regarding the effectiveness of the use of fluorescence-guided surgery in patients with tumors located in eloquent areas of the brain, in which resection can be stopped because the high risk of an increase of neurological deficit. Objective is to evaluate the effectiveness of fluorescence-guided surgery in patients with tumors located near the motor cortex and corticospinal tract. Methods. Our research includes 108 adult patients with gliomas near the motor cortex and corticospinal tract divided into two groups depending on the use of fluorescence-guided surgery: control (34 patients without fluorescence) and main (62 patients with fluorescence). Results. There is no difference between study groups in the radicality of surgery and neurological outcomes. Conclusion. Fluorescence-guided surgery can be a useful tool as part of complex intraoperative monitoring in patients with tumors located near the motor cortex and corticospinal tract despite the absence of a statistically significant difference.

Indocyanine Green (ICG) Fluorescence-Assisted Open Surgery Using the Rubina® Lens System in the Pediatric Population: A Single-Center Prospective Case Series.

There are scarce papers about the use of fluorescence-guided surgery (FGS) in the open surgical field. This study aimed to assess the usefulness of FGS in an open setting in the pediatric population and to report our preliminary experience using the Rubina® Lens system. All patients undergoing ICG fluorescence-assisted open surgery over the period September 2022-September 2023 were enrolled. Each surgical procedure was performed using the Rubina® Lens for ICG fluorescence visualization. A total of 25 patients, 14 boys and 11 girls with a median age at surgery of 5.8 years-old (range 0-15), were enrolled. Surgical indications were dermoid/epidermoid cysts of the head (n = 7), lymphangiomas of the head/neck (n = 2), thyroglossal duct cysts (n = 7), gynecomastia (n = 3), preauricular fistula (n = 2), second branchial cleft fistula (n = 1), fibrolipoma of the shoulder (n = 1) and myofibroma of the gluteal/perineal region (n = 2). In all procedures, an intralesional injection of 2.5 mg/mL ICG solution using a 30-gauge needle was administered. No adverse reactions to ICG occurred. Median operative time was 68.6 min (range 35-189). The visualization of ICG-NIRF with the Rubina® Lens was achieved in all cases. No intraoperative complications were reported. Postoperative complications occurred in 3/25 patients (12%), with gynecomastia (n = 1), thyroglossal duct cyst (n = 1) and neck lymphangioma (n = 1), who developed a fluid collection in the surgical site, requiring needle aspiration in outpatient care (Clavien-Dindo 2). Complete mass excision was confirmed with pathology reports. Based on this initial experience, FGS using the Rubina® Lens was very helpful in open surgery, providing enhanced visualization of anatomy and identification of margins, real-time reliability and low complication rate. It was easy to use, time saving, feasible and clinically safe. Previous experience in MIS is necessary to adopt this technology. The accuracy of the injection phase is important to avoid diffusion of the ICG into the perilesional tissue.

IndoCyanine Green fluorescence guided resections in hepatobiliary surgery

Background: Fluorescence-guided surgery (FGS) has recently gained popularity as a promising technique for treating visceral, hepatobiliary, and pancreatic neoplasms. It involves using laser sources to illuminate injected substances that emit a fluorescence signal, guiding surgical procedures, and providing real-time visualization of otherwise undetectable structures. This review explores the advancements in hepatobiliary surgery using IndoCyanine Green (ICG) fluorescence guided resections. Methods: The review examined the use of FGS in identifying subcapsular liver tumors, millimetric hepatocellular carcinoma, intrahepatic cholangiocarcinoma, liver metastases, and various benign liver neoplasms. In addition, fluorescence cholangiography using ICG injection was explored to improve liver surgery's accuracy and safety. Results: The review found that ICG fluorescence-guided resections can potentially improve surgical outcomes by enhancing the accuracy and safety of procedures. The use of fluorescence cholangiography allows for the efficient identification of the bile ducts and helps surgeons avoid damaging critical structures during liver surgery. Conclusion: ICG fluorescence-guided resections represent a promising method for improving surgical outcomes and patient safety for visceral and hepatobiliary Surgery. It is a quick, easy, inexpensive, and safe device that can be used for various surgical applications. As imaging systems continue to improve, fluorescence imaging can become a widely used intraoperative navigation tool for open, laparoscopic, and robotic surgery.

Immunohistochemical Evaluation of Candidate Biomarkers for Fluorescence-Guided Surgery of Myxofibrosarcoma Using an Objective Scoring Method.

Myxofibrosarcoma (MFS) is the most common soft-tissue sarcoma subtype in elderly patients. Local recurrence (LR) remains a major concern as the lack of intraoperative guidance and an infiltrative growth pattern with long, slender tails hamper surgeons' ability to achieve adequate resection margins for MFS. Fluorescence-guided surgery (FGS) could overcome this concern by delineating tumor tissue during surgery. One of the most important steps to successful FGS is to define a tumor-specific biomarker that is highly overexpressed in tumor tissue while low or absent in adjacent healthy tissue. The aim of this study is to evaluate the expression of eight previously selected promising biomarkers for FGS in MFS tissue samples with adjacent healthy tissue using immunohistochemistry (IHC). The following eight biomarkers were stained in seventeen paraffin-embedded MFS samples: tumor endothelial marker-1 (TEM-1, also known as endosialin/CD248), vascular endothelial growth factor receptor-1 (VEGFR-1, also known as Flt-1), vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Flk1), vascular endothelial growth factor-A (VEGF-A), epidermal growth factor receptor (EGFR), insulin-like growth factor-1 receptor (IGF-1R), platelet derived growth factor receptor-α (PDGFR-α), and cluster of differentiation 40 (CD40, also known as TNFRSF5). A pathologist specializing in sarcoma annotated the margin between the tumor and adjacent healthy tissue in each MFS tissue sample. Subsequently, we used an objective IHC scoring method to assess and compare the difference in staining intensity between the tumor and adjacent healthy tissue, which is crucial for the use of FGS. TEM-1, VEGF-A, and PDGFR-α stained all MFS tumors, while the other biomarkers did not show expression in all MFS tumors. Ultimately, TEM-1 was identified as the most suitable biomarker for FGS in MFS based on higher tumor-to-background (TBR) staining intensity compared to VEGF-A and PDGFR-α, regardless of preoperative therapy. TEM-1-targeted FGS tracers should be further investigated to optimize MFS treatment.

Can fluorescence-guided surgery improve optimal surgical treatment for ovarian cancer? A systematic scoping review of clinical studies

BackgroundThe predicament of achieving optimal surgical intervention faced by surgeons in treating ovarian cancer has driven research into improving intra-operative detection of cancer using fluorescent materials.ObjectiveTo provide a literature overview...

Spontaneous intracerebral haemorrhage secondary to 5-ALA-induced thrombocytopaenia in a paediatric patient: case report and literature review

IntroductionThe primary objective of neurosurgical management of malignant gliomas is maximal safe resection of the tumour. One of the main obstacles in achieving this is the ability to accurately discriminate between tumour edges and the surrounding healthy brain tissue. The use of fluorescence-guided surgery utilising 5-aminolevulinic acid (5-ALA), first introduced more than 20 years ago, has become an invaluable adjunct in high-grade glioma surgery in adults. However, as 5-ALA is not licensed for use in paediatric patients, the safety profile for such use remains undetermined.Case reportWe describe the case of a 4-year-old boy who underwent 5-ALA-guided resection of a fourth ventricle anaplastic ependymoma. Although complete resection was achieved and the patient awoke from surgery well with no neurological deficits, the patient developed acute transaminitis, anaemia, thrombocytopaenia and coagulopathy postoperatively. The patient had a sudden neurological deterioration on postoperative day 2; imaging revealed that he had suffered a spontaneous right frontal intracerebral haemorrhage. The patient returned to theatre for surgical decompression and evacuation of the haematoma, and ultimately went on to make a full recovery.ConclusionThe use of 5-ALA in paediatric patients can be helpful in maximising surgical resection, but the associated safety profile remains undefined. Further research is urgently warranted in order to characterise the efficacy and risk of the use of 5-ALA in the paediatric population.

Use of fluorescence image-guided surgery and autofluorescence in thyroid and parathyroid surgery

Fluorescence-guided surgery (FGS) has seen increased interest in recent decades. Technological advances have made it more widely accessible for a variety of applications, including thyroid and parathyroid surgery. Parathyroid autofluorescence can be utilized to help identify parathyroid glands during thyroid or parathyroid surgery and reduce rates of postoperative hypocalcemia after thyroidectomy. Fluorescent dyes such as indocyanine green (ICG) may be used to evaluate perfusion of parathyroid glands during thyroid or parathyroid surgery and help guide decision-making about auto-transplantation or which gland to leave as a remnant. As an emerging technology, additional research is needed to determine the optimal use of FGS in thyroid and parathyroid surgery, including the developing field of molecularly targeted fluorophores. FGS is an exciting and promising field that may help make endocrine surgery safer, faster, and more effective.

Mind the gap—the use of sodium fluoresceine for resection of brain metastases to improve the resection rate

Introduction and purposeBrain metastases appear to be well resectable due to dissectable tumor margins, but postoperative MRI quite often depicts residual tumor with potential influence on tumor control and overall survival. Therefore, we introduced sodium fluoresceine into the routine workflow for brain metastasis resection. The aim of this study was to evaluate whether the use of fluorescence-guided surgery has an impact on postoperative tumor volume and local recurrence.Material and methodsWe retrospectively included patients who underwent surgical resection for intracranial metastases of systemic cancer between 11/2017 and 05/2021 at our institution. Tumor volumes were assessed pre- and postoperatively on T1-CE MRI. Clinical and epidemiological data as well as follow-up were gathered from our prospective database.ResultsSeventy-nine patients (33 male, 46 female) were included in this study. Median preoperative tumor volume amounted to 11.7cm3 and fluoresceine was used in 53 patients (67%). Surgeons reported an estimated gross total resection (GTR) in 95% of the cases, while early postoperative MRI could confirm GTR in 72%. Patients resected using fluoresceine demonstrated significantly lower postoperative residual tumor volumes with a difference of 0.7cm3 (p = 0.044) and lower risk of local tumor recurrence (p = 0.033). The use of fluorescence did not influence the overall survival (OS). Postoperative radiotherapy resulted in a significantly longer OS (p = 0.001).DiscussionWhile GTR rates may be overrated, the use of intraoperative fluorescence may help neurosurgeons to achieve a more radical resection. Fluoresceine seems to facilitate surgical resection and increase the extent of resection thus reducing the risk for local recurrence.

Narrative review on applications of fluorescence-guided surgery in adult and paediatric urology

: More recently, there has been a substantial increase in the use of fluorophores to guide open and laparoscopic procedures with the ultimate aim to optimize both oncological and functional outcomes and to reduce operative time and blood loss. Fluorescent dyes are currently adopted to allow safer and more accurate dissections during both oncological and reconstructive procedures thanks to their ability of highlighting tumour margins and tissue vascularization. Urology is one of the fields in which fluorescence-guided surgery (FGS) has proved to be most useful. In particular, it has assumed a pivotal role in the surgical treatment of oncological patients affected by kidney, bladder, prostate and penis cancers and in the management of paediatric urological conditions. This review aims to provide an update on the use of FGS in adult and paediatric urology, drawing attention to its most recent and interesting applications in this very innovative field of research. Although FGS has been only recently introduced in the clinical scenario, it can be already considered a powerful tool to improve oncological, anatomical and functional outcomes in both adult and paediatric urology. An increased identification of lymph nodes (LNs), a more accurate visualization of tumour margins and a better definition of blood supply and lymphatic drainage have proved to be greatly beneficial for patients undergoing urological procedures, whether they are adults or children. Longitudinal studies with larger sample sizes are still needed to draw firm conclusions and to confirm its benefits in urology.

Effects of the tumour microenvironment on protoporphyrin IX accumulation in glioblastoma

Abstract Aims Glioblastoma is the most common primary brain tumour and has a poor prognosis. Standard clinical intervention involves the resection of the tumour volume, chemotherapy and radiotherapy. However, achieving gross-total resection is challenging due to poorly defined boundaries as a result of tumour infiltration. Fluorescence-guided surgery (FGS) utilises an apparently selective accumulation of protoporphyrin IX (PPIX) that occurs in areas of glioblastoma after systemic administration of the metabolite 5-aminolevulinic acid (5-ALA). We have investigated the metabolic basis for the heterogeneity of the PPIX fluorescent signal, and its implications for glioma biology. Method Using glioblastoma cell lines and patient-derived primary cells, we have monitored the uptake of 5-ALA and conversion to the fluorescent molecule PPIX. Stable isotope tracing coupled with GCMS and LCMS was used to analyse intra- and extracellular metabolite levels arising from exogenous 5-ALA administration under both normoxic (21% O2) and hypoxic (1% O2) conditions. Results Uptake of exogenous 5-ALA from culture media and conversion to PPIX is observed in a time and dose-dependent manner in both normoxia and hypoxia. High levels of PPIX accumulation are associated with reduced cell proliferation despite the majority of the PPIX synthesised not being retained within the tumour cell, but exported into the medium. Under hypoxic conditions, reduced fluorescence is observed as a result of the decrease in oxygen availability likely affecting the oxygen-dependent enzymes. Stable isotope tracing experiments indicate an increase in the glutamine-derived succinate pool in response to exogenous 5-ALA, which is dependent on flux through the heme pathway. Conclusion Our data suggest that different microenvironments within the tumour alter the activity of the heme biosynthetic pathway, resulting in differential fluorescence in glioblastoma. It paves the way by which we could work to alter the glioblastoma microenvironment in order to further improve the use of FGS in guiding surgery across these devastating tumours.

Real-time fluorescence image-guided gastrointestinal oncologic surgery: Towards a new era.

Technological improvements are crucial in the evolution of surgery. Real-time fluorescence-guided surgery (FGS) has spread worldwide, mainly because of its usefulness during the intraoperative decision-making processes. The success of any gastrointestinal oncologic resection is based on the anatomical identification of the primary tumor and its regional lymph nodes. FGS allows also to evaluate the blood perfusion at the gastrointestinal stumps after colorectal or esophageal resections. Therefore, a reduction on the anastomotic leak rates has been postulated as one of the foreseeable benefits provided by the use of FGS in these procedures. Although the use of fluorescence in lymph node detection was initially described in breast cancer surgery, the technique is currently applied in gastric or splenic flexure cancers, as they both present complex and variable lymphatic drainages. FGS allows also to perform intraoperative lymphograms or sentinel lymph node biopsies. New applications of FGS are being developed to assist in the detection of peritoneal metastases or in the evaluation of the tumor resection margins. The present review aims to provide a general overview of the current status of real-time FGS in gastrointestinal oncologic surgery. We put a special focus on the different applications of FGS, discussing the main findings and limitations found in the contemporary literature and also the promising near future applications.

BIMG-17. EFFECTS OF THE TUMOUR MICROENVIRONMENT ON PROTOPORPHYRIN IX ACCUMULATION IN GLIOBLASTOMA

Glioblastoma is the most common primary brain tumour and has a poor prognosis. The median survival is less than two years despite clinical intervention that usually involves the resection of the tumour volume, chemotherapy and radiotherapy. Achieving gross-total resection is challenging due to poorly defined boundaries as a result of tumour infiltration. Fluorescence-guided surgery (FGS) utilises an apparently selective accumulation of protoporphyrin IX (PPIX) that occurs in areas of glioblastoma after administration of the metabolite, 5-aminolevulinic acid (5-ALA). 5-ALA and the fluorescent metabolite, PPIX, sit within the endogenous heme biosynthetic pathway, which suggests that FGS is not only an important clinical tool, but also highlights differing metabolic phenotypes naturally present throughout the tumour. Genetic and mechanistic studies into this phenomenon have shown that differential expression of metabolite transporters, altered activity of the heme pathway enzymes and variable nutrient availability are all factors in the accumulation of PPIX. However, little is known about the cellular driving force for the uptake of 5-ALA and subsequent conversion into PPIX. Our data suggest that different microenvironments within the tumour alter the activity of the heme biosynthetic pathway, resulting in differential fluorescence in glioblastoma. It paves the way for work to alter the glioblastoma microenvironment in order to further improve the use of FGS in guiding surgery across these devastating tumours.

Use of intraoperative fluorescence to enhance robot-assisted radical prostatectomy.

Robot-assisted radical prostatectomy has become the standard of care for the removal of localized prostate cancer. Positive outcomes depend upon the precise removal of the prostate and associated tissue without damage to nearby structures. This process can be aided by fluorescence-guided surgery to enhance the visual contrast between different structures. Herethe authors have conducted a systematic review using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines to identify ten investigations into the use of fluorescence-guided surgery in robot-assisted radical prostatectomy. These studies used fluorescent tracers to identify structures, including the prostate, neurovascular bundle and lymph nodes. These studies demonstrate the safe and effective use of fluorescence-guided surgery in robot-assisted radical prostatectomy and pave the way for further developments in this field.

Abstract PS3-04: Breast tumor targeted fluorescence guidance for intraoperative margin assessment

Abstract BACKGROUND: A majority of the 270,000 women diagnosed with breast cancer in the United States annually are candidates for breast conserving surgery (lumpectomy), however, up to 30% of patients require reoperation due to positive margins. The ideal resolution is to make residual in-breast disease visible at the time of surgery. The goal of this research is to develop a systemically administered fluorescent molecular imaging agent that is targeted to breast tumors for intraoperative margin assessment using fluorescence guided surgery (FGS). METHODS: To identify targetable cell-surface markers for use in FGS, gene expression profiling of publicly available Affymetrix mRNA microarray data from patients with invasive ductal or invasive lobular carcinoma tumors (n=34) and surrounding normal breast tissues (n=32) was performed. Protein expression of select targets was determined by immunohistochemistry (IHC) staining of patient tissue specimens. A humanized CEACAM6 (carcinoembryonic cell adhesion molecule 6) specific antibody fragment (scFv-Fc [IgG4]) was conjugated to the near-infrared IRDye800CW fluorescent dye (LiCor) by NHS-ester linkage to create CEACAM6-800. The conjugate was characterized for in vitro specificity by immunocytochemistry (ICC) and live cell uptake studies. As proof-of-principle, the conjugate (15 µg) was intravenously injected into nude mice bearing orthotopic human MCF-7 breast cancer mammary fat pad (MFP) xenograft tumors with endogenous expression of CEACAM6. FGS was performed using the “SurgVision” clinical imaging platform (approved for clinical use in Europe) at 24 h post-injection.RESULTS: We identified 263 genes with higher mRNA expression in tumor relative to normal. From this list, we selected 9 genes for confirmation of protein expression by IHC. Of the 9 targets, only CEACAM6 had no protein expression in normal breast tissues and robust expression in ~50% of tumors. ICC staining using the CEACAM6-800 conjugate and multiple human ductal carcinoma tumor cell lines confirmed the cell surface localization of CEACAM6. In live-cell uptake studies, CEACAM6-800 accumulated on the cell surface as early as 10 min after addition to the media and was internalized after 90 min. Blocking studies demonstrated specificity. In the proof-of-principal FGS study, we observed high fluorescence in MCF-7 MFP tumors relative to background and the tumors and their residuals were completely removed in the first attempt. CONCLUSION: We have confirmed CEACAM6 protein as a cell-surface adhesion receptor target for breast cancer FGS. The CEACAM6-800, fluorescent-dye to antibody-fragment conjugate was prepared and tested in vitro for sub-cellular localization and specific uptake. In vivo studies demonstrated specific uptake into MFP tumors and FGS for tumor resection. We are currently performing studies to determine the in vivo pharmacokinetics of CEACAM6-800 uptake and clearance in tumor and normal tissues and pre-clinical post-surgery survival using our pre-clinical models. These studies will enable future translational studies. Citation Format: Narges Tafreshi, Mikalai Budzevich, Weihong Sun, Daruka Mahadevan, Marie Catherine Lee, David L Morse. Breast tumor targeted fluorescence guidance for intraoperative margin assessment [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS3-04.

Fluorescence-guided surgical management of medication-related osteonecrosis of the jaws.

Medication-related osteonecrosis of the jaw (MRONJ) results in progressive destruction of the jawbones, and advanced cases demand surgical intervention. The total removal of necrotic bone is required to prevent recurrence and infection. However, determining the limits between necrotic and healthy bone is a challenge. The use of fluorescence to detect tissue alterations and determine necrosis extension is a promising method to avoid inadequate bone debridement. In the literature, there are several studies and reported cases that successfully use fluorescence-guided surgery (FGS) of MRONJ. The objective of this study was to present a critical review of the literature regarding the intraoperative use of optic fluorescence to differentiate healthy and necrotic bone in MRONJ. Studies that evaluated the intraoperative use of optic fluorescence to determine the surgical margins of MRONJ were searched in Pubmed/Medline and Scopus databases using the following terms: "medication-related osteonecrosis of the jaws" and "fluorescence". Eighteen papers describing the intraoperative use of FGS in ONJ management were selected, totalizing 218 patients. Of those 18 papers, only 5 were prospective studies, and the other 13 were isolated case reports, case series, and technical notes. The use of FGS to delimitate the resection margin of MRONJ is a promising method. There is no need for the application of exogenous fluorophore to perform FGS and the most often used light device was the VELScope® system. Further prospective studies with larger samples are still required to ascertain the fluorescence validity as a supporting method in MRONJ surgical treatment and to establish clinical protocols.

Indocyanine green matching phantom for fluorescence-guided surgery imaging system characterization and performance assessment

.Significance: Expanded use of fluorescence-guided surgery with devices approved for use with indocyanine green (ICG) has led to a range of commercial systems available. There is a compelling need to be able to independently characterize system performance and allow for cross-system comparisons.Aim: The goal of this work is to expand on previous proposed fluorescence imaging standard designs to develop a long-term stable phantom that spectrally matches ICG characteristics and utilizes 3D printing technology for incorporating tissue-equivalent materials.Approach: A batch of test targets was created to assess ICG concentration sensitivity in the 0.3- to 1000-nM range, tissue-equivalent depth sensitivity down to 6 mm, and spatial resolution with a USAF test chart. Comparisons were completed with a range of systems that have significantly different imaging capabilities and applications, including the Li-Cor® Odyssey, Li-Cor® Pearl, PerkinElmer® Solaris, and Stryker® Spy Elite.Results: Imaging of the ICG-matching phantoms with all four commercially available systems showed the ability to benchmark system performance and allow for cross-system comparisons. The fluorescence tests were able to assess differences in the detectable concentrations of ICG with sensitivity differences >10× for preclinical and clinical systems. Furthermore, the tests successfully assessed system differences in the depth-signal decay rate, as well as resolution performance and image artifacts. The manufacturing variations, photostability, and mechanical design of the tests showed promise in providing long-term stable standards for fluorescence imaging.Conclusions: The presented ICG-matching phantom provides a major step toward standardizing performance characterization and cross-system comparisons for devices approved for use with ICG. The developed hybrid manufacturing platform can incorporate long-term stable fluorescing agents with 3D printed tissue-equivalent material. Further, long-term testing of the phantom and refinements to the manufacturing process are necessary for future implementation as a widely adopted fluorescence imaging standard.

Considerations for NIR-I and short-wave infrared (SWIR) fluorescence imaging within a clinical operating room.

Short-wave infrared (SWIR/NIR-II) fluorescence imaging has received increased attention for use in fluorescence-guided surgery (FGS) due to the potential for higher resolution imaging of subsurface structures and reduced autofluorescence compared to conventional NIR-I imaging. As with any fluorescence imaging modality introduced in the operating room, an appropriate accounting of contaminating background signal from other light sources in the operating room is an important step. Herein, we report the background signals in the SWIR and NIR-I emitted from commonly-used equipment in the OR, such as ambient and operating lights, LCD screens and surgical guidance systems. These results can guide implementation of protocols to reduce background signal.

Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models

IntroductionClaudins are tight-junction proteins, which maintain an epithelial barrier in normal colon cells. Overexpression of Claudin-1 has been implicated for development of colon cancer. We postulated that Claudin-1 may be a useful target in near-infrared imaging and fluorescence-guided surgery. MethodsWe conjugated Claudin-1 antibody to LI-COR IR800DyeCW (Claudin-1-IRDye800CW). Western blotting of 9 human colon cancer cell lysates was performed. Animal imaging was performed with the LI-COR Pearl Trilogy Fluorescence Imaging System. A dose-response study was carried out with subcutaneous LS174T colon cancer cell line models. Increasing doses of Claudin-1-IRDye800CW via tail vein injection were administered to three groups of mice. Two groups of mice were used as controls (antibody alone, and dye alone). In vivo imaging was performed at 24, 48, and 72 h after administration of the conjugated dye. Orthotopic implantation of patient-derived tumors and cell lines was performed and peritoneal carcinomatosis models were created. After tumor growth, mice were administered Claudin-1-IRDye800CW and imaged in vivo 48 h later. The mice were euthanized and laparotomy was performed to assess internal organs and toxicity. ResultsWestern blotting revealed that all colon cancer cell lysates expressed varying amounts of Claudin-1. All tumors demonstrated strong and specific fluorescence labeling at 800 nm, even with the lowest dose of 12.5 μg of Claudin-1-IRDye800CW. ConclusionsClaudin-1 is a useful target for near-infrared antibody-based imaging for visualization of colorectal tumors for future use in fluorescence-guided surgery.

Fluorescence guided surgery in liver tumors: applications and advantages.

The use of fluorescence-guided surgery for benign and malignant hepatobiliary (HPB) neoplasms has significantly increased and improved imaging methods creating new interesting perspectives. A major challenge in HPB surgery is performing radical resection with maximal preservation of the liver parenchyma and obtaining a low rate of complications. Despite the developments, visual inspection, palpation, and intraoperative ultrasound remain the most utilized tools during surgery today. In laparoscopic and robotic HPB surgery palpation is not possible. Fluorescence imaging enables identification of subcapsular liver tumors through accumulation of indocyanine green (ICG), after preoperative intravenous injection, in cancerous tissues of hepatocellular carcinoma and in noncancerous hepatic parenchyma, around intrahepatic cholangiocarcinoma and liver metastases, and it can also be used for visualizing extrahepatic bile duct anatomy and hepatic segmental borders, increasing the accuracy and the easiness of open and minimally invasive hepatectomy. (www.actabiomedica.it)

Near-infrared–conjugated humanized anti-carcinoembryonic antigen antibody targets colon cancer in an orthotopic nude-mouse model

BackgroundThe success of a curative surgery for cancer is dependent on the complete removal of all cancer cells. Tumor visualization by the surgeon can be enhanced through fluorescent-antibody targeting. To further develop such technology, we selected humanized anti-carcinoembryonic antigen (CEA) conjugated to a near-infrared dye to target orthotopically-implanted human colon cancer in nude mice. Materials and methodsThe HT-29 human colon cancer cell line was grown in culture and subcutaneously injected in mice. After 3 wk of growth, tumors were resected and cut into 2 mm3 fragments that were sutured to the cecum of five additional nude mice for orthotopic implantation. The tumors were allowed to grow for 4 wk at which point 3 had successful orthotopic tumor growth and were selected for injection of the humanized anti-CEA antibody conjugated to the near-infrared dye IRDye800CW (anti-CEA-IRDye800CW). The antibody-dye conjugate (75 μg) was administered via tail vein injection. Images were obtained with the Pearl Trilogy Small Animal Imaging System with both 700 and 800 nm channels and evaluated using Image Studio. ResultsLaparotomy was performed 24 h after labeling the tumors. When imaged through the 800 nm channel, the tumors were observed to be strongly labeled with anti-CEA-IRDye800. At 48 h, laparotomy was repeated which again demonstrated strong labeling of the tumors through the 800 nm channel, but with a lower absolute intensity (in relative units), than at 24 h. ConclusionsHumanized anti-CEA-IRDye800CW can rapidly and effectively label CEA-expressing human colon cancer in an orthotopic nude mouse model. Given the ability of this technology to target and label tumors with great specificity, the anti-CEA-IRDye800CW is currently being developed for clinical use in fluorescence-guided surgery.