Catheter-based in vivo imaging of enzyme activity and gene expression: feasibility study in mice

PURPOSE: To construct and evaluate an interventional catheter-based imaging system for intravital monitoring of molecularly sensitive near-infrared fluorescent probes and optical marker genes. MATERIALS AND METHODS: An imaging device that was based on a miniaturized fiberoptic sensor (MIFS) was built in which images created with a 2.7-F fiberoptic catheter were relayed through a dichroic mirror, through a bandpass filter, and on two independent cameras. This system permitted simultaneous recording of white-light and fluorescent images. Spatial resolution, spectral transmissions, and sensitivity were determined in vitro. In vivo testing was performed in nude mice bearing intraperitoneal tumors that express green fluorescent protein and in a mouse model of ovarian carcinoma with enzyme-activatable near-infrared probes sensitive to tumoral protease activity. Signal intensity on images of tumors and that on images of normal tissue were measured and compared with t test. RESULTS: The catheter, which was advanced through an 18-gauge sheath, showed resolution of 7 line pairs per millimeter and detection limit for fluorochrome Cy5.5 of 1-10 pmol. Detection of endogenous green fluorescent protein gene expression was feasible in tumor nodules smaller than 1 mm in diameter (mean tumor signal intensity, 153.26 +/- 26.45 [SD], compared with that of adjacent nontumoral tissue of 36.73 +/- 11.69; P <.008). Similarly, activation of the near-infrared probe by tumoral proteases could be detected in peritoneal tumor seeds of ovarian cancer model with mean tumor signal intensity of 246.33 +/- 7.77 compared with that of adjacent nontumoral tissue of 41.56 +/- 18.64 (P <.001). Mean contrast-to-noise ratio in the near-infrared channel exceeded white-light contrast-to-noise ratio by a factor of 6.7 (P <.02). CONCLUSION: With this system, in vivo MIFS imaging of gene expression, enzyme activity, and potentially other molecular events is feasible, through direct interventional access to several organs and body cavities and potentially through transvascular approaches.     

Imaging of differential protease expression in breast cancers for detection of aggressive tumor phenotypes

PURPOSE: To determine if different expression levels of tumor cathepsin-B activity in well differentiated and undifferentiated breast cancers could be revealed in vivo with optical imaging. MATERIALS AND METHODS: A well differentiated human breast cancer (BT20, n = 8) and a highly invasive metastatic human breast cancer (DU4475, n = 8) were implanted orthotopically in athymic nude mice. Tumor-bearing animals were examined in vivo with near-infrared fluorescence (NIRF) imaging 24 hours after intravenous injection of an enzyme-sensing imaging probe. Immunohistochemistry, Western blotting (on cells and whole tumor samples), and correlative fluorescence microscopy were performed. RESULTS: Both types of breast cancers activated the NIRF probe so that tumors became readily detectable. However, in tumors of equal size, there was a 1.5-fold higher fluorescence signal in the highly invasive breast cancer (861 arbitrary units +/- 88) compared with the well differentiated lesion (566 arbitrary units +/- 36, P <.01). Western blotting confirmed a higher cathepsin-B protein content in the highly invasive breast cancer (DU4475) of about 1.4-fold (whole tumor samples) to 1.7-fold (cells). Immunohistochemistry and fluorescence microscopy findings confirmed the imaging findings. CONCLUSION: Cathepsin-B enzyme activity can be determined in vivo with NIRF optical imaging, while differences in tumoral expression may correlate with tumor aggressiveness.     

Dual optical and nuclear imaging in human melanoma xenografts using a single targeted imaging probe

INTRODUCTION: Dual-labeled imaging agents that allow both nuclear and optical imaging after a single injection would be advantageous in certain applications. In this study, we synthesized and characterized a dual-labeled RGD (Arg-Gly-Asp) peptide and compared nuclear and optical images obtained with this agent. METHODS: 111In-DTPA-Lys(IRDye800)-c(KRGDf) composed of both the 111In chelator diethylenetriaminepentaacetic acid (DTPA) and the near-infrared (NIR) fluorescent dye IRDye800 (excitation/emission, 765/792 nm) was synthesized. The probe was characterized with regard to in vitro biological activity and in vivo pharmacokinetics and the ability to target integrin alphavbeta3. Tumors of mice injected with the dual-labeled probe were imaged both by gamma scintigraphy and NIR fluorescence optical camera. RESULTS: DTPA-Lys(IRDye800)-c(KRGDf), DTPA-Lys-c(KRGDf) and c(KRGDf) inhibited the adhesion of melanoma M21 cells to vitronectin-coated surface with the similar biological activity. Both 111In-DTPA-Lys(IRDye800)-c(KRGDf) and 111In-DTPA-Lys-c(KRGDf) had significantly higher uptakes in alphavbeta3-positive M21 melanoma than in alphavbeta3-negative M21-L melanoma at 4-48 h after their injection. Side-by-side comparison of images obtained using 111In-DTPA-Lys(IRDye800)-c(KRGDf) revealed that in living mice, both optical imaging and gamma scintigraphy enabled noninvasive detection of the bound probe to alphavbeta3-positive tumors, with optical images providing improved resolution and sensitive detection of the superficial lesions and gamma images providing sensitive detection of deeper structures. CONCLUSION: The dual-labeled imaging probe 111In-DTPA-Lys(IRDye800)-c(KRGDf) was found to specifically bind to alphavbeta3 in melanoma tumor cells. Employing both nuclear and optical imaging with a single imaging probe may facilitate translation of NIR fluorescence optical imaging into clinical applications.     

Near-infrared optical imaging of protease activity for tumor detection

PURPOSE: To build and test an optical imaging system that is sensitive to near-infrared fluorescent molecular probes activated by specific enzymes in tumor tissues in mice. MATERIALS AND METHODS: The imaging system consisted of a source that delivered 610-650-nm excitation light within a lighttight chamber, a 700-nm longpass filter for selecting near-infrared fluorescence emission photons from tissues, and a charge-coupled device (CCD) for recording images. The molecular probe was a biocompatible autoquenched near-infrared fluorescent compound that was activated by tumor-associated proteases for cathepsins B and H. Imaging experiments were performed 0-72 hours after intravenous injection of the probe in nude mice that bore human breast carcinoma (BT-20). RESULTS: The imaging system had a maximal spatial resolution of 60 microns, with a field of view of 14 cm2. The detection threshold of the nonquenched near-infrared fluorescent dye was subpicomolar in the imaging phantom experiments. In tissue, 250 pmol of fluorochrome was easily detected during the 10-second image acquisition. After intravenous injection of the probe into the tumor-bearing animals, tumors as small as 1 mm became detectable because of tumor-associated enzymatic activation of the quenched compound. CONCLUSION: Tumor proteases can be used as molecular targets, allowing visualization of millimeter-sized tumors. The development of this technology, probe design, and optical imaging systems hold promise for molecular imaging, cancer detection, and evaluation of treatment.     

In vitro and in vivo investigation of matrix metalloproteinase expression in metastatic tumor models

INTRODUCTION: Overexpression of matrix metalloproteinases (MMPs), particularly MMP-2 and MMP-9, has been correlated with poor prognosis in several cancer types including lung, colon and breast. Noninvasive detection of MMP expression might allow physicians to better determine when more aggressive cancer therapy is appropriate. The peptide CTT (CTTHWGFTLC) was identified as a selective inhibitor of MMP-2/9 that inhibits the growth of MDA-MB-435 human breast cancer xenografts. METHODS: CTT was conjugated with the bifunctional chelator DOTA (1,4,7,10-tetraazacyclotetradecane-N,N',N',N'-tetraacetic acid) for radiolabeling with (64)Cu (t(1/2)=12.7 h, 17.4% beta(+), 39% beta(-)), a radionuclide suitable for positron emission tomography (PET). In vitro affinity was determined in a fluorogenic substrate assay. Tumor gelatinase targeting was evaluated in both biodistribution and microPET imaging studies. RESULTS: Cu(II)-DOTA-CTT inhibited hMMP-2 (EC(50)=8.7 microM) and mMMP-9 (EC(50)=18.2 microM) with similar affinity to CTT (hMMP-2 EC(50)=13.2 microM; mMMP-9 EC(50)=11.0 microM). In biodistribution and microPET imaging studies, (64)Cu-DOTA-CTT was taken up by MMP-2/9-positive B16F10 murine melanoma tumors. Subsequently, imaging studies using (64)Cu-DOTA-CTT were performed on MDA-MB-435 tumor-bearing mice. With zymography, tumor MMP-2/9 expression in this model was shown to be inconsistent, resulting in microPET detection of the MDA-MB-435 tumor in only 1 of 24 imaged mice. Following limited imaging success, (64)Cu-DOTA-CTT was shown to have poor in vivo stability. CONCLUSIONS: Despite some evidence for selective uptake of (64)Cu-DOTA-CTT by gelatinase-expressing tumors, the low affinity for MMP-2 and MMP-9 and in vivo instability make this an inadequate radioligand for in vivo tumor evaluation.     

Colonic adenocarcinomas: near-infrared microcatheter imaging of smart probes for early detection--study in mice

PURPOSE: To prospectively evaluate the ability of micro-fiberoptic catheters, which simultaneously record white light and near-infrared (NIR) images, to reveal colonic neoplasms after the intravenous administration of activatable "smart" probes that increase in NIR fluorescence subsequent to protease activation. MATERIALS AND METHODS: The institutional animal care committee approved all animal experiments. CT26 tumor cells were orthotopically implanted into the descending colon of C57BL6/J mice (n=10). Thirteen days later, mice intravenously received either 2 nmol of a protease-sensing probe that had cathepsin B as a major activator (n=5) or saline (control animals [n=5]). One day later, animals were noninvasively examined to the point of the splenic flexure by using microcatheter imaging. Excised colons were subsequently evaluated with epifluorescence imaging, histologic examination, and cathepsin B immunohistochemistry. Student t test was used for statistical analysis, with P<.05 considered to indicate a significant difference. RESULTS: Results with fiberoptic imaging demonstrated that all tumors were visible with the protease-activatable probe, even when they were not readily apparent at white light imaging. A target-to-background ratio (TBR) of 8.86 for tumor to adjacent normal mucosa was achieved in the NIR channel after probe administration (P=.001), whereas white light images resulted in a TBR of 1.14 (P>.5) based on luminosity. The tumoral NIR fluorescence intensity was more than 30-fold greater in probe-injected animals than in control animals, indicating that essentially all of the signal recorded in lesions was from activatable probe administration. Results of immunohistochemistry confirmed cathepsin B overexpression in the tumor compared with adjacent mucosa. CONCLUSION: The use of NIR imaging microcatheters combined with protease-activatable smart probes results in a beacon effect that highlights tumors with high TBRs; this technique thus may be a potentially useful adjunct to white light colonoscopy in the future.     

Dual-function probe for PET and near-infrared fluorescence imaging of tumor vasculature.

To date, the in vivo imaging of quantum dots (QDs) has been mostly qualitative or semiquantitative. The development of a dual-function PET/near-infrared fluorescence (NIRF) probe can allow for accurate assessment of the pharmacokinetics and tumor-targeting efficacy of QDs. METHODS: A QD with an amine-functionalized surface was modified with RGD peptides and 1,4,7,10-tetraazacyclodocecane-N,N',N',N'-tetraacetic acid (DOTA) chelators for integrin alpha(v)beta(3)-targeted PET/NIRF imaging. A cell-binding assay and fluorescence cell staining were performed with U87MG human glioblastoma cells (integrin alpha(v)beta(3)-positive). PET/NIRF imaging, tissue homogenate fluorescence measurement, and immunofluorescence staining were performed with U87MG tumor-bearing mice to quantify the probe uptake in the tumor and major organs. RESULTS: There are about 90 RGD peptides per QD particle, and DOTA-QD-RGD exhibited integrin alpha(v)beta(3)-specific binding in cell cultures. The U87MG tumor uptake of (64)Cu-labeled DOTA-QD was less than 1 percentage injected dose per gram (%ID/g), significantly lower than that of (64)Cu-labeled DOTA-QD-RGD (2.2 +/- 0.3 [mean +/- SD] and 4.0 +/- 1.0 %ID/g at 5 and 18 h after injection, respectively; n = 3). Taking into account all measurements, the liver-, spleen-, and kidney-to-muscle ratios for (64)Cu-labeled DOTA-QD-RGD were about 100:1, 40:1, and 1:1, respectively. On the basis of the PET results, the U87MG tumor-to-muscle ratios for DOTA-QD-RGD and DOTA-QD were about 4:1 and 1:1, respectively. Excellent linear correlation was obtained between the results measured by in vivo PET imaging and those measured by ex vivo NIRF imaging and tissue homogenate fluorescence (r(2) = 0.93). Histologic examination revealed that DOTA-QD-RGD targets primarily the tumor vasculature through an RGD-integrin alpha(v)beta(3) interaction, with little extravasation. CONCLUSION: We quantitatively evaluated the tumor-targeting efficacy of a dual-function QD-based probe with PET and NIRF imaging. This dual-function probe has significantly reduced potential toxicity and overcomes the tissue penetration limitation of optical imaging, allowing for quantitative targeted imaging in deep tissue.     

Quantitative real-time catheter-based fluorescence molecular imaging in mice

PURPOSE: To prospectively evaluate an optical imaging system designed to perform quantitative, intravital catheter-based imaging of fluorescent molecular probes. MATERIALS AND METHODS: This study was performed according to a protocol approved by the institutional animal care committee. A fiberoptic catheter imaging system was developed to implement a normalization algorithm for real-time quantitative near-infrared (NIR) imaging. The system was validated with in vitro imaging of fluorochrome phantoms and in vivo fluorescence measurements obtained in tumors implanted in murine abdomens (n = 7) after administration of an enzyme-activatable NIR probe. Standard analysis of variance tests were used to determine significant dissimilarities in signal from distinct fluorochrome concentrations. The clinical utility of the system was further evaluated by imaging orthotopically implanted murine colonic adenocarcinomas (n = 4). RESULTS: Raw NIR fluorescence intensities, which were measured with a fiberoptic catheter placed above wells of varying NIR fluorochrome concentration, varied markedly (>100%) with catheter position, while the corrected NIR signal was confined to a range of values within 10% of their mean for each individual fluorochrome concentration and were significantly distinct (P < .001) between relevant concentration ranges. Similar results were observed for the in vivo measurements from the abdominally implanted tumors, with raw NIR signal varying 20% from the mean and corrected NIR signal varying only 1% from the mean. The colonic studies revealed that the correction method was robust enough for use during minimally invasive imaging procedures. CONCLUSION: The authors have developed and implemented a method for quantitative real-time catheter-based fluorescence imaging that resolves NIR signal dependence on changes in catheter position.     

Near-Infrared Fluorescence Imaging of Matrix Metalloproteinase 2 Activity as a Biomarker of Vascular Remodeling in Hemodialysis Access

Purpose

To establish the capability of near-infrared fluorescence (NIRF) imaging for the detection of matrix metalloproteinase 2 (MMP-2) activity as a biomarker of vascular remodeling (VR) in arteriovenous fistulae (AVFs) in vivo.

宽视场荧光内镜的构建及其初步应用

 目的 为探索胃癌分子成像技术的新方法,构建一套新型灵活的宽视场荧光内镜系统,并用于裸鼠胃癌移植瘤模型荧光成像研究。方法 通过合理的光路设计及光学组件机加工,装配宽视场荧光内镜系统,通过对不同浓度的2-DG-750荧光探针进行成像,分析系统性能;对尾静脉注射100 μl 2-DG-750荧光探针的裸鼠胃癌移植瘤模型进行在体荧光成像,对ROI的像素值和成像图进行分析。结果 体外成像ROI的像素值与荧光探针浓度存在较好的线性回归关系,在2-DG-750剂量为31.3 pmol时,该系统即可检测到荧光信号,提示该系统灵敏性好;在体成像显示用该系统联合2-DG-750荧光探针观察到了肿瘤组织的较高荧光信号强度。结论 宽视场荧光内镜系统具有很好的灵敏性,可以同时实现白光及荧光的双模式成像,具有实现光学分子成像诊断胃癌的潜在应用价值。     

Recurrent glomus tumors of fingertips: MR imaging evaluation

PURPOSE: To determine the magnetic resonance (MR) imaging findings in recurrent glomus tumors of the fingertips. MATERIALS AND METHODS: Twenty-four consecutive patients with recurrent pain after previous excision of a glomus tumor of the fingertip underwent MR imaging studies and surgery. T1-weighted spin-echo MR images were obtained in each patient before and after intravenous injection of contrast material; T2-weighted spin-echo and three-dimensional gradient-recalled echo images were also obtained. MR angiography was performed in four patients. Postsurgical histopathologic analysis revealed recurrent glomus tumors in 22 patients. Signal intensity, enhancement, and margins of the scar tissue and the recurrent tumors at MR were assessed. RESULTS: The postsurgical scars were depicted in 21 (88%) of 24 patients with all sequences but were best demonstrated on gradient-recalled echo MR images. Seven patients had undergone multiple surgical procedures and had extensive scar tissue and, in one case, a neuroma. In all patients, MR imaging revealed a nodule compatible with the diagnosis of a recurrent glomus tumor. In 13 (54%) of 24 patients, the nodule had typical features of a glomus tumor. In eight (33%) of 24 patients, the tumors had low signal intensity or isointensity compared with the nail bed on T2-weighted images. In six (25%) of 24 patients, the tumors had faint enhancement after intravenous gadolinium chelate administration. The margins of the tumors were blurred by scar tissue in nine of 24 cases. CONCLUSION: MR imaging can aid in the evaluation of recurrent glomus tumors.     

[<Superscript>18</Superscript>F]FBEM-Z<Subscript>HER2:342</Subscript>-Affibody molecule—a new molecular tracer for in vivo monitoring of HER2 expression by positron emission tomography

Purpose The expression of human epidermal growth factor receptor-2 (HER2) receptors in cancers is correlated with a poor prognosis. If assessed in vivo, it could be used for selection of appropriate therapy for individual patients and for monitoring of the tumor response to targeted therapies. We have radiolabeled a HER2-binding Affibody molecule with fluorine-18 for in vivo monitoring of the HER2 expression by positron emission tomography (PET). Materials and methods The HER2-binding Z_HER2:342–Cys Affibody molecule was conjugated with N-2-(4-[¹⁸F]fluorobenzamido)ethyl]maleimide ([¹⁸F]FBEM). The in vitro binding of the resulting radioconjugate was characterized by receptor saturation and competition assays. For in vivo studies, the radioconjugate was injected into the tail vein of mice bearing subcutaneous HER2-positive or HER2-negative tumors. Some of the mice were pre-treated with non-labeled Z_HER2:342−Cys. The animals were sacrificed at different times post-injection, and the radioactivity in selected tissues was measured. PET images were obtained using an animal PET scanner. Results In vitro experiments indicated specific, high-affinity binding to HER2. PET imaging revealed a high accumulation of the radioactivity in the tumor as early as 20 min after injection, with a plateau being reached after 60 min. These results were confirmed by biodistribution studies demonstrating that, as early as 1 h post-injection, the tumor to blood concentration ratio was 7.5 and increased to 27 at 4 h. Pre-saturation of the receptors with unlabeled Z_HER2:342–Cys lowered the accumulation of radioactivity in HER2-positive tumors to the levels observed in HER2-negative ones. Conclusion Our results suggest that the [¹⁸F]FBEM-Z_HER2:342 radioconjugate can be used to assess HER2 expression in vivo.     

ICG纳米探针在早期结肠癌诊断中的应用及其价值

 

目的 评估ICG纳米探针在结肠癌荧光分子成像中的靶向性、荧光效应及其早期诊断价值。方法 建立裸鼠结肠癌皮下移植瘤模型,应用人血清白蛋白(HSA)包裹的ICG纳米探针,并以Folate RsenseTM680、吲哚菁绿作为对照组,经裸鼠结肠癌皮下移植瘤模型尾静脉注射探针后,进行活体荧光分子成像,观察HAS-ICG纳米探针的成像效果,量化分析肿瘤部位的荧光信号强度。结果 活体荧光分子成像结果显示:探针HAS-ICG、Folate RsenseTM680均在实验瘤鼠皮下肿瘤部位出现浓聚,浓聚高峰分别在注射后1 h、24 h。与对照组相比,HAS-ICG纳米探针比商业探针有较好的靶向标记性和信噪比。结论 HSA-ICG纳米探针具有很好的标记HCT116结肠肿瘤细胞的能力,可通过荧光分子成像技术诊断早期裸鼠结肠癌变。

     

Near-infrared fluorescence imaging of HER-2 protein over-expression in tumour cells

The aim of this study was to evaluate in vitro and in vivo imaging of HER-2-over-expressing tumours using near-infrared optical imaging. A fluorochrome probe was designed by coupling Cy5.5 to anti-HER-2 antibodies. Cells over-expressing (SK-BR-3 cells) or normally expressing (PE/CA-PJ34 cells) the HER-2 protein were incubated with the probe. After removing unbound probe molecules, fluorescence intensities were determined (a.u.: arbitrary units). Cells were additionally investigated using FACS and laser scanning microscopy. The probe was also injected intravenously into tumours bearing SK-BR-3 (n=3) or PE/CA-PJ34 (n=3). Whole-body fluorescence images were generated and analysed. The incubation of SK-BR-3 cells with the probe led to higher fluorescence intensities [2,133 (±143) a.u.] compared to controls [975 (±95) a.u.]. The results from FACS and immunocytochemical analysis were in agreement with these findings. A distinct dependency between the fluorescence intensity and the cell number used in the incubations was detected. In vivo, the relative fluorescence intensities in SK-BR-3 tumours were higher than in PE/CA-PJ34 tumours at 16–24 h after probe application. HER-2-over-expressing tumours were depictable in their original size. Labelling of HER-2 with Cy5.5 is suitable for in vitro and in vivo detection of HER-2-over-expressing tumour cells.     

Optical imaging of spontaneous breast tumors using protease sensing 'smart' optical probes

OBJECTIVE: The objective of this study was to determine if spontaneous breast cancer lesions can be detected by fluorescence reflectance imaging (FRI) and fluorescence mediated tomography (FMT) using protease-sensing optical probes. MATERIALS AND METHODS: Transgenic (FVB/N-TgN (WapHRAS)69Lin Y)) mice, which spontaneously develop breast cancer, were injected intravenously with a cathepsin-sensing fluorescent imaging probe. FRI and FMT were performed 24 hours after probe injection and region of interest (ROI) analysis was performed. Magnetic resonance images were acquired for anatomic coregistration with the FMT data. Moreover, correlative immunohistochemistry and fluorescence microscopy were performed. RESULTS: All tumor nodules were clearly delineated by FRI showing an average signal intensity of 380 +/- 106 AU. Similarly, tumors were clearly detected by FMT imaging. Immunohistochemistry confirmed cathepsin-B expression of primary tumors and fluorescence microscopy revealed a strong Cy 5.5 deposition in the tissue. CONCLUSIONS: FRI and FMT using "smart" protease sensing probes permits detection of experimental spontaneous breast cancers. Because the expression levels of various proteases correlate with patient outcome, this technique may not only help to detect, but also to differentiate breast cancers noninvasively.     

颅脑血管外皮细胞瘤的CT、MRI与病理对照研究

目的：探讨颅脑血管外皮细胞瘤的临床、病理和ＣＴ、ＭＲＩ特征。材料与方法：收集经ＣＴ和ＭＲＩ诊断为脑膜瘤,经手术病理证实为血管外皮细胞瘤１２例,年龄３４￣５８岁。ＣＴ检查７例,均采用增强前、后常规扫描。ＭＲＩ检查１２例,采用ＳＥ序列增强前、后扫描。结果：１２例肿瘤全部发生在颅内脑外,７例ＣＴ平扫呈低等混合密度２例,呈等高混合密度５例;增强后扫描呈不均匀强化６例,均匀强化１例。骨窗显示病灶局部侵蚀性骨质破坏４例。ＭＲＩ平扫,Ｔ１ＷＩ呈等高不均匀信号１０例,呈等信号２例;Ｔ２ＷＩ呈不均匀等高信号１０例,呈等信号２例。１２例中７例显示脑膜尾征;增强后扫描１ 均呈不均匀强化。结论：血管外皮细胞瘤具有一定的ＣＴ、ＭＲＩ特征性表现：分叶状、丰富的血管流空、肿瘤内密度或信号不均匀、无肿瘤内钙化和骨质增生、局部颅骨呈溶骨性破坏。     

Contrast-enhanced near-infrared laser mammography with a prototype breast scanner: feasibility study with tissue phantoms and preliminary results of imaging experimental tumors

RATIONALE AND OBJECTIVES: Near-infrared (NIR) optical mammography without contrast has a low specificity. The application of optical contrast medium may improve the performance. The concentration-dependent detectability of a new NIR contrast medium was determined with a prototype optical breast scanner. In vivo imaging of experimental tumors was performed. METHODS: The NIR contrast agent NIR96010 is a newly synthesized, hydrophilic contrast agent for NIR mammography. A concentration-dependent contrast resolution was determined for tissue phantoms consisting of whole milk powder and gelatin. A central part of the phantoms measuring 2 x 2 cm2 without contrast was replaced with phantom material containing 1 micromol/L to 25 nmol/L NIR96010. The composite phantoms were measured with a prototype NIR breast scanner with lasers of lambda1 = 785 nm and lambda2 = 850 nm wavelength. Intensity profiles and standard deviations of the transmission signal in areas with and without contrast were determined by linear fit procedures. Signal-to-noise ratios and spatial resolution as a function of contrast concentration were determined. Near-infrared imaging of five tumor-bearing SCID mice (MX1 breast adenocarcinoma, tumor diameter 5-10 mm) was performed before and after intravenous application of 2 micromol/kg NIR96010. RESULTS: Spectrometry showed an absorption maximum of the contrast agent at 755 nm. No spectral shifts occurred in protein-containing solution. Signal-to-noise ratio in the transmission intensity profiles ranged from 1.1 at 25 nmol/L contrast to 28 at 1 micromol/L. At concentrations <40 nmol/L, no differentiation from the background was possible. The transitional area between the contrast-free edge of the phantom and the central contrast-containing part appeared in the profiles as a steep increase with a width of 4.2 +/- 1.8 mm. The experimental tumors were detectable in nonenhanced images as well as contrast-enhanced images, with better delineation after contrast administration. In postcontrast absorption profiles, a 44.1% +/- 11.3% greater absorption increase was seen in tumor tissue compared with normal tissue. CONCLUSIONS: The laser wavelength lambda1 of the prototype laser mammography device was not situated at maximum absorption of the contrast agent NIR96010 but on the descending shoulder of the absorption spectrum. This implies a 20% signal loss for contrast detection. Despite the nonideal measurement conditions, concentrations as low as 40 nmol/L were detectable in vitro. In vivo, all tumors were detectable in color-coded nonenhanced scans as well as in contrast-enhanced scans, with better delineation after contrast administration.     

裸鼠胃癌移植瘤的近红外荧光成像

目的：通过对非特异性探针Cy5.5在裸鼠体内分布及显像研究,探讨近红外荧光成像对胃癌的早期诊断及动态监测价值。方法：用MGC-803细胞株建立胃癌动物模型,进行早期、实时活体及离体显像实验。结果：近红外荧光成像可检测早期胃癌移植瘤的平均大小为2.807mm×3.045mm,与游标卡尺测得的肿瘤大小呈直线相关,r=0.924,P〈0.05。Cy5.5主要分布在肿瘤组织,主要代谢器官为肾脏;注入探针30min后,裸鼠肿瘤部位成像清晰,荧光寿命、荧光强度均高于对照部位（P〈0.05）。60min后,肿瘤区的荧光强度始高于血液（P〈0.05）。90min时达峰值。肿瘤部位的平均荧光寿命为（3.1376±0.9894）ns,明显高于对侧部位（P〈0.05）。结论：近红外荧光成像可用于胃癌的早期诊断及瘤体的动态监测。     

阔韧带平滑肌瘤的MR诊断

目的:探讨阔韧带平滑肌瘤的MR表现及诊断价值.方法:回顾性分析13例经手术和病理证实为阔韧带平滑肌瘤的MR表现.结果:13例中肿块位于子宫右侧4例,左侧8例,两侧1例.肿块呈圆形、类圆形10例,不规则形或分叶状3例.肿块大于5 cm占84.6%(11/13).T1WI肿块均呈等或略低信号;T2WI为中等信号,其中11例肿块中见不均匀斑点状、梭条状高信号影,2例见大片状液化坏死灶.注射钆喷替酸葡甲胺(Gd-DTPA)后扫描肿块实质部分明显强化,显示包膜完整,与子宫等周围脏器分界清晰10例,包膜不清,与子宫有粘连且分界不清3例;包膜不强化且显示更清晰.结论:阔韧带平滑肌瘤的MR表现具有一定的特征性:盆腔内、子宫外大而有完整包膜的实性或实性为主的肿块,T1WI呈等或略低信号,T2WI呈中等信号,其内常可见不均匀斑点状、梭条状高信号影,增强扫描肿瘤强化明显.MR具有良好的软组织分辨率、多方位成像的特点,有利于肿瘤结构、成份与包膜显示,对明确诊断有肯定的价值.     

Transarterial Delivery of a Biodegradable Single-Agent Theranostic Nanoprobe for Liver Tumor Imaging and Combinatorial Phototherapy

PurposeTo assess selective accumulation of biodegradable nanoparticles within hepatic tumors after transarterial delivery for in vivo localization and combinatorial phototherapy.Materials and MethodsA VX2 hepatic tumor model was used in New Zealand white rabbits. Transarterial delivery of silicon naphthalocyanine biodegradable nanoparticles was performed using a microcatheter via the proper hepatic artery. Tumors were exposed via laparotomy, and nanoparticles were observed by near-infrared (NIR) fluorescence imaging. For phototherapy, a handheld NIR laser (785 nm) at 0.6 W/cm² was used to expose tumor or background liver, and tissue temperatures were assessed with a fiberoptic temperature probe. Intratumoral reactive oxygen species formation was assessed using a fluorophore (2′,7′-dichlorodihydrofluorescein diacetate).ResultsNanoparticles selectively accumulated within viable tumor by NIR fluorescence. Necrotic portions of tumor did not accumulate nanoparticles, consistent with a vascular distribution. NIR-dependent heat generation was observed with nanoparticle-containing tumors, but not in background liver. No heat was generated in the absence of NIR laser light. Reactive oxygen species were formed in nanoparticle-containing tumors exposed to NIR laser light, but not in background liver treated with NIR laser or in tumors in the absence of NIR light.ConclusionsBiodegradable nanoparticle delivery to liver tumors from a transarterial approach enabled selective in vivo tumor imaging and combinatorial phototherapy.