scholarly journals Multi-Modal PET and MR Imaging in the Hen’s Egg Test-Chorioallantoic Membrane (HET-CAM) Model for Initial In Vivo Testing of Target-Specific Radioligands

Cancers ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 1248
Author(s):  
Gordon Winter ◽  
Andrea B. F. Koch ◽  
Jessica Löffler ◽  
Mika Lindén ◽  
Christoph Solbach ◽  
...  
Keyword(s):  
Animal Experiments ◽  
Chorioallantoic Membrane ◽  
Xenograft Model ◽  
In Vivo Model ◽  
Pet Tracer ◽  
Hen’S Egg ◽  
Novel Target ◽  
Cam Model ◽  
Proof Of Principle

The validation of novel target-specific radioligands requires animal experiments mostly using mice with xenografts. A pre-selection based on a simpler in vivo model would allow to reduce the number of animal experiments, in accordance with the 3Rs principles (reduction, replacement, refinement). In this respect, the chick embryo or hen’s egg test–chorioallantoic membrane (HET-CAM) model is of special interest, as it is not considered an animal until day 17. Thus, we evaluated the feasibility of quantitative analysis of target-specific radiotracer accumulation in xenografts using the HET-CAM model and combined positron emission tomography (PET) and magnetic resonance imaging (MRI). For proof-of-principle we used established prostate-specific membrane antigen (PSMA)-positive and PSMA-negative prostate cancer xenografts and the clinically widely used PSMA-specific PET-tracer [68Ga]Ga-PSMA-11. Tracer accumulation was quantified by PET and tumor volumes measured with MRI (n = 42). Moreover, gamma-counter analysis of radiotracer accumulation was done ex-vivo. A three- to five-fold higher ligand accumulation in the PSMA-positive tumors compared to the PSMA-negative tumors was demonstrated. This proof-of-principle study shows the general feasibility of the HET-CAM xenograft model for target-specific imaging with PET and MRI. The ultimate value for characterization of novel target-specific radioligands now has to be validated in comparison to mouse xenograft experiments.

3D monitoring of tumor volume in an in vivo model

2020 ◽  
Vol 76 (2) ◽  
pp. 123-131
Author(s):  
Johannes Troebs ◽  
Claudia Asam ◽  
Eric Pion ◽  
Lukas Prantl ◽  
Thiha Aung ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Real Time ◽  
Xenograft Model ◽  
Chemotherapeutic Agents ◽  
In Vivo Model ◽  
Primary Tumors ◽  
Tumor Weight ◽  
Digital Microscope ◽  
Cam Model

BACKGROUND: The ability to evaluate tumor development within experimental oncology is of upmost importance. However, determining tumor volumes in 3D in vivo tumor models is challenging. The chick chorioallantoic membrane (CAM) model represents an optimized xenograft model that surpasses many disadvantages that are inherent to rodent models and provides the opportunity of real-time monitoring of tumor growth. OBJECTIVE: The objective of this study was to introduce a new method that enables monitoring of tumor growth within the CAM model throughout the course of the experiment. METHODS: Sarcoma cell lines and sarcoma primary tumors were grafted onto the CAM of fertilized chicken eggs. A digital microscope (Keyence VHX-6000) was used for 3D volume monitoring before and after tumor excision and compared it to tumor weight. RESULTS: Accuracy of tumor volumes was validated through correlation with tumor weight. In and ex ovo tumor volumes correlated significantly with tumor weight values. CONCLUSIONS: The described method can be used to assess the effects of chemotherapeutic agents on the growth of tumors that have been grafted onto the CAM and further advance personalized cancer therapy. In summary, we established a promising protocol that enables in vivo real-time tracking of tumor growth in the CAM model using a digital microscope.

The chick embryo chorioallantoic membrane model: a research approach for ex vivo and in vivo experiments

2021 ◽  
Vol 28 ◽  
Author(s):  
Ana Isabel Fraguas-Sánchez ◽  
Cristina Martín-Sabroso ◽  
Ana Isabel Torres-Suárez
Keyword(s):  
Animal Models ◽  
Chorioallantoic Membrane ◽  
New Drugs ◽  
Biological Research ◽  
Research Areas ◽  
Chick Chorioallantoic Membrane ◽  
Biodistribution Studies ◽  
Toxicological Studies ◽  
Cam Model

Background: The chick chorioallantoic membrane (CAM) model has attracted a great deal of interest in pharmaceutical and biological research as an alternative or complementary in vivo assay to animal models. Traditionally, CAM assay has been widely used to perform some toxicological studies, specifically to evaluate the skin, ocular and embryo toxicity of new drugs and formulations, and perform angiogenesis studies. Due to the possibility to generate the tumors onto the CAM, this model has also become an excellent strategy to evaluate the metastatic potential of different tumours and test the efficacy of novel anticancer therapies in vivo. Moreover, in the recent years, its use has considerably grown in other research areas, including the evaluation of new anti-infective agents, the development of biodistribution studies and tissue engineering research. Objectives: This manuscript provides a critical overview of the use of CAM model in pharmaceutical and biological research, especially to test the toxicity of new drugs and formulations and the biodistribution and the efficacy of novel anticancer and anti-infective therapies, analyzing its advantages and disadvantages compared to animal models. Conclusion: The chick chorioallantoic membrane model shows great utility in several research areas, such as cancer, toxicology, biodistribution studies and anti-infective therapies. In fact, it has become an intermediate stage between in vitro experiments and animal studies, and, in the case of toxicological studies (skin and ocular toxicity), has even replaced the animal models.

scholarly journals Investigating the ocular toxicity potential and therapeutic efficiency of in situ gel nanoemulsion formulations of brinzolamide

2020 ◽  
Vol 9 (4) ◽  
pp. 578-587
Author(s):  
Sima Talaei ◽  
Mohammad Mehdi Mahboobian ◽  
Mojdeh Mohammadi
Keyword(s):  
Intraocular Pressure ◽  
Membrane Surface ◽  
Chorioallantoic Membrane ◽  
In Situ Gel ◽  
Toxicity Potential ◽  
Vessel Injury ◽  
Hen’S Egg

Abstract Glaucoma is an ocular disease i.e. more common in older adults with elevated intraocular pressure and a serious threat to vision if it is not controlled. Due to the limitations regarding the conventional form of brinzolamide (Azopt®), two optimum formulations of in situ gel nanoemulsion were developed. To ensure the safety and efficacy of developed formulations for ocular drug delivery, the current study was designed. MTT assay was carried out on the human retinal pigmentation epithelial cells. To investigate the irritation potential of the chosen formulations, hen’s egg test-chorioallantoic membrane as a borderline test between in vivo and in vitro methods has been done. The modified Draize method was utilized to evaluate eye tolerance against the selected formulations. Intraocular pressure was measured by applying the prepared formulations to the eyes of normotensive albino rabbits in order to assess the therapeutic efficacy. Based on MTT test, cell viability for NE-2 at 0.1% and NE-1 at 0.1 and 0.5% concentrations was acceptable. The results of the hen’s egg test-chorioallantoic membrane test indicated no sign of vessel injury on the chorioallantoic membrane surface for both formulations. Also, during 24 h, both formulations were well-tolerated by rabbit eyes. The pharmacodynamics effects of formulations had no difference or were even higher than that of suspension in case of adding lower concentration (0.5%) of brinzolamide to the formulations. With regard to the results of the mentioned methods, our advanced formulations were effective, safe, and well-tolerated, thus can be introduced as an appropriate vehicle for ocular delivery of brinzolamide.

scholarly journals In-Vivo Studies of the Angiogenic Potential of Mandur Bhasma

2021 ◽  
pp. 79-84
Author(s):  
Ekta Tomar ◽  
Sonali Wairagade ◽  
Arachana Gharote ◽  
Ranjit S. Ambad ◽  
Dhruba Hari Chandi
Keyword(s):  
Experimental Study ◽  
Analytical Study ◽  
Chorioallantoic Membrane ◽  
In Vivo Studies ◽  
Classical Literature ◽  
Standard Drug ◽  
Angiogenic Potential ◽  
Egg Shell ◽  
Cam Model

Background: Mandur Bhasma is a herbo-mineral compound. It is prepared by Putapaka method. It is described as Raktasanjanan. In the current study, Mandur Bhasma was prepared with a standardized method w.s.r to Rasatarangini and an experimental study was done to observe the Angiogenic property of Mandur Bhasma. The current study will analyze angiogenic potential of Mandur Bhasma using chick CAM model. This research is intended to study the possible role of Mandur Bhasma on angiogenesis and establishing properties of Mandur Bhasma as an angiogenic by newer means. The experimental study inside the egg shell will be carried out on a membrane known as “chorioallantoic membrane”. Objectives: To Prepare Mandur Bhasma Physicochemical and Analytical study of Mandur Bhasma To verify the angiogenic potential of Mandur bhasma using the chicken chorioallantoic membrane (CAM) model. To compare Angeogenic potential of Mandur bhasma with standard drug progesterone Methodology: Relevant classical literature regarding Mandur will be reviewed and the data will be collected. Mandur Shodhan with Gomutra and Mandur Maran with Triphala decoction will be done. Analytical Study like Organoleptic Test for Rasa, Gandha, Varna, Sparsha, Physicochemical Tests and other analytical test like ICP-AES /ICPMS, XRD structure of Bhasma, EDAX-NANO Particle Size will be done. Expected Results: Changes will be observed in objective outcomes. Conclusion: Conclusion will be drawn by suitably analyzing data.

scholarly journals LncRNA DLEU1 Contributes to the Growth and Invasion of Colorectal Cancer via Targeting miR-320b/PRPS1

2021 ◽  
Vol 11 ◽  
Author(s):  
Dong Xu ◽  
Fei Yang ◽  
Yongchao Fan ◽  
Wanling Jing ◽  
Jianfei Wen ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
Cell Apoptosis ◽  
Human Cancer ◽  
Xenograft Model ◽  
Underlying Mechanism ◽  
Migration And Invasion ◽  
Phosphoribosyl Pyrophosphate ◽  
Novel Target

Growing evidences suggest that long non-coding RNAs (lncRNAs) are closely correlated to the development of human cancer, such as colorectal cancer (CRC). A previous report suggested that DLEU1 accelerated CRC development. However, DLEU1’s underlying mechanism in CRC remains unclear. In our study, the level of DLEU1 in CRC tissues is investigated by qRT-PCR. Our data exhibited that DLEU1 level was observably increased in CRC tissues and CRC cell lines and was closely associated with bad prognosis of CRC patients. CRC cell proliferation was repressed by sh-LncRNA DLEU1, whereas cell apoptosis was markedly stimulated. Moreover, knockdown of DLEU1 inhibited cell migration and invasion. Mechanistically, through interacting with miR-320b in CRC, DLEU1 promoted the level of PRPS1 which was a target of miR-320b. The rescue experiment confirmed that knockdown of DLEU1 repressed cell proliferation, migration and invasion while stimulated cell apoptosis via miR-320b/phosphoribosyl pyrophosphate synthetase 1 (PRPS1) axis. Meanwhile, the data of xenograft model exhibited that inhibition of DLEU1 suppressed tumor growth in vivo. In summary, DLEU1 knockdown may repress PRPS1 expression via miR-320b, and then repress cell proliferation, migration and invasion while stimulate cell apoptosis. Our research may provide a novel target for the treatment of CRC.

scholarly journals In Situ DNA/Protein Interaction Assay to Visualize Transcriptional Factor Activation

2020 ◽  
Vol 3 (4) ◽  
pp. 80
Author(s):  
Michela Corsini ◽  
Emanuela Moroni ◽  
Cosetta Ravelli ◽  
Elisabetta Grillo ◽  
Marco Presta ◽  
...  
Keyword(s):  
Protein Interaction ◽  
Nuclear Translocation ◽  
Chorioallantoic Membrane ◽  
Camp Response Element Binding ◽  
Transcriptional Factors ◽  
In Vivo Model ◽  
Element Binding Protein ◽  
Dna Protein Interaction

The chick embryo chorioallantoic membrane (CAM) represents a powerful in vivo model to study several physiological and pathological processes including inflammation and tumor progression. Nevertheless, the possibility of deepening the molecular processes in the CAM system is biased by the absence/scarcity of chemical and biological reagents, designed explicitly for avian species. This is particularly true for transcriptional factors, proteinaceous molecules that regulate various cellular responses, including proliferation, survival, and differentiation. Here, we propose a detailed antibody-independent protocol to visualize the activation and nuclear translocation of transcriptional factors in cells or in tissues of different animal species. As a proof of concept, DNA/cAMP response element-binding protein (CREB) interaction was characterized on the CAM tissue using oligonucleotides containing the palindromic binding sequence of CREB. Scrambled oligonucleotides were used as controls. In situ DNA/protein interaction protocol is a versatile method that is useful for the study of transcription factors in the cell and tissue of different origins.

scholarly journals Degradation of Mitochondria and Oxidative Stress as the Main Mechanism of Toxicity of Pristine Graphene on U87 Glioblastoma Cells and Tumors and HS-5 Cells

2019 ◽  
Vol 20 (3) ◽  
pp. 650 ◽  
Author(s):  
Sławomir Jaworski ◽  
Barbara Strojny ◽  
Ewa Sawosz ◽  
Mateusz Wierzbicki ◽  
Marta Grodzik ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Chorioallantoic Membrane ◽  
In Vivo Model ◽  
Pristine Graphene ◽  
Infrared Measurements ◽  
The Impact

Due to the development of nanotechnologies, graphene and graphene-based nanomaterials have attracted immense scientific interest owing to their extraordinary properties. Graphene can be used in many fields, including biomedicine. To date, little is known about the impact graphene may have on human health in the case of intentional exposure. The present study was carried out on U87 glioma cells and non-cancer HS-5 cell lines as in vitro model and U87 tumors cultured on chicken embryo chorioallantoic membrane as in vivo model, on which the effects of pristine graphene platelets (GPs) were evaluated. The investigation consisted of structural analysis of GPs using transmission electron microscopy, Fourier transmission infrared measurements, zeta potential measurements, evaluation of cell morphology, assessment of cell viability, investigation of reactive oxygen species production, and investigation of mitochondrial membrane potential. The toxicity of U87 glioma tumors was evaluated by calculating the weight and volume of tumors and performing analyses of the ultrastructure, histology, and protein expression. The in vitro results indicate that GPs have dose-dependent cytotoxicity via ROS overproduction and depletion of the mitochondrial membrane potential. The mass and volume of tumors were reduced in vivo after injection of GPs. Additionally, the level of apoptotic and necrotic markers increased in GPs-treated tumors.

Study on the Patency of the Contralateral Iliac Vein After Stenting Across the Iliocaval Confluence With an Experimental In Vivo Model

2019 ◽  
Vol 53 (8) ◽  
pp. 644-648 ◽  
Author(s):  
Xicheng Zhang ◽  
Wennuo Huang ◽  
Huiming Yu ◽  
Yong Chen ◽  
Jiaxin Liu ◽  
...  
Keyword(s):  
Doppler Ultrasonography ◽  
Histopathological Examination ◽  
Animal Experiments ◽  
Vena Cava ◽  
Iliac Vein ◽  
In Vivo Model ◽  
Cross Sectional ◽  
Stenosis Rate ◽  
Stent Strut

Objective: Stenting is the preferred treatment for iliac vein lesions. For the treatment of occlusions in the junction of the iliac vein and the inferior vena cava (IVC), the stent needs to be positioned in the IVC to cover the lesion. However, the pathological changes in the contralateral iliac vein due to stent coverage on its ostium remain unclear. We observed the patency of the contralateral iliac vein via animal experiments. Methods: The stents were placed in the left iliac vein and extended into the IVC in 8 beagle dogs. Doppler ultrasonography, angiography, and histopathological examination were used to assess the patency and histopathological changes in the contralateral iliac vein. Results: Angiography showed patency of the contralateral iliac vein and no sign of thrombosis or stenosis. Twelve months after stenting, Doppler ultrasonography showed a stenotic change in the ostium of the contralateral iliac vein. The histopathological examination showed that the stent strut at the ostium of the contralateral iliac vein was mostly covered by the intima, and the cross-sectional stenosis rate was greater than 60%. Conclusions: The coverage of the iliac vein stent on the ostium of the contralateral iliac vein does not cause complete occlusion of the contralateral vein but can cause significant stenosis at the ostium of the contralateral iliac vein, which is considered to be a potential risk factor for thrombosis.

Comparing surgical tissue versus biopsy tissue in the development of a clear cell renal cell carcinoma xenograft model.

2016 ◽  
Vol 34 (2_suppl) ◽  
pp. 519-519
Author(s):  
Yiyu Dong ◽  
Brandon Manley ◽  
A. Ari Hakimi ◽  
Jonathan A. Coleman ◽  
Paul Russo ◽  
...  
Keyword(s):  
Xenograft Model ◽  
In Vivo Model ◽  
P Value ◽  
Cell Renal Cell Carcinoma ◽  
Immunodeficient Mice ◽  
Biopsy Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Selection Of

519 Background: The use of xenograft tumor models is considered the ideal platform to investigate the effects and toxicities of novel drugs in primary human tumors. The establishment of a personalized xenograft model using preoperative or pretherapy biopsy for patients with metastatic or high risk disease could improve selection of targeted therapy. We report on our xenograft model using various tissue sources including biopsies and correlation with patient’s clinical features. Methods: 56 specimens from primary and metastatic ccRCC from 48 patients were collected. After surgery (n=35) or biopsy (n=21) the specimen was transplanted either subcutaneously or after cell culture to immunodeficient mice. Tumor engraftment was followed for up to 4 months. Successfully engrafted patient-derived tumors were passaged to further mice. Conformation of xenograft tumors with formalin-fixed, paraffin-embedded and Hematoxylin and eosin stained tumor sections was done to assure morphological concordance with the patients tumor. We used a two-tailed two proportion z-test to compare the number of successful xenografts harvested from surgical tissue or biopsy tissue. Results: Overall 25 of the 56 specimens were successful in growing tumor in our immunodeficient mice. The frequency of success based on the type and site of tissue harvest may be seen in Table 1. We found biopsy tissue to be significantly more successful compared to surgical tissue, 61.9% compared to 34.2% (p-value=0.044). Conclusions: We believe our xenograft model, using biopsy tissue, demonstrates the feasibility of a real time personalized in vivo model to aid in the selection of targeted treatments for systemic therapy in ccRCC patients. [Table: see text]

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