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Article ; Online: Fc engineering: serum half-life modulation through FcRn binding.

Olafsen, Tove

Methods in molecular biology (Clifton, N.J.)

2012 Volume 907, Page(s) 537–556

Abstract: Controlling the half-life of pharmaceuticals through Fc engineering is a desirable approach to achieve optimal exposure and targeting. The long serum residence time of gamma immunoglobulins is attributed to the Fc binding to the neonatal Fc receptor ( ... ...

Abstract	Controlling the half-life of pharmaceuticals through Fc engineering is a desirable approach to achieve optimal exposure and targeting. The long serum residence time of gamma immunoglobulins is attributed to the Fc binding to the neonatal Fc receptor (FcRn). The residues in the Fc region that interact with FcRn have been mapped and individual mutations of these residues have demonstrated reduced affinity to FcRn and faster blood clearance. Here, we describe site-specific mutagenesis of Fc residues in a scFv-Fc fusion protein, as well as the mammalian production, purification, characterization, and the in vivo pharmacokinetics of these antibody fragments.
MeSH term(s)	Animals ; Chromatography, Affinity ; Chromatography, Thin Layer ; Half-Life ; Histocompatibility Antigens Class I/genetics ; Histocompatibility Antigens Class I/isolation & purification ; Histocompatibility Antigens Class I/metabolism ; Humans ; Iodine Radioisotopes ; Mice ; Mutagenesis, Site-Directed ; Protein Binding ; Protein Engineering/methods ; Receptors, Fc/genetics ; Receptors, Fc/isolation & purification ; Receptors, Fc/metabolism ; Recombinant Fusion Proteins/isolation & purification ; Recombinant Fusion Proteins/pharmacokinetics ; Serum/metabolism
Chemical Substances	Fc receptor, neonatal ; Histocompatibility Antigens Class I ; Iodine Radioisotopes ; Receptors, Fc ; Recombinant Fusion Proteins
Language	English
Publishing date	2012
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-61779-974-7_31
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: 89Zr-ImmunoPET for the Specific Detection of EMP2-Positive Tumors.

Chan, Ann M / Olafsen, Tove / Tsui, Jessica / Salazar, Felix B / Aguirre, Brian / Zettlitz, Kirstin A / Condro, Michael / Wu, Anna M / Braun, Jonathan / Gordon, Lynn K / Ashki, Negin / Whitelegge, Julian / Xu, Shili / Ikotun, Oluwatayo / Lee, Jason Thanh / Wadehra, Madhuri

Molecular cancer therapeutics

2024

Abstract: Epithelial membrane protein-2 (EMP2) is upregulated in a number of tumors and therefore remains a promising target for monoclonal antibody (mAb)-based therapy. In the current study, image guided therapy for an anti-EMP2 mAb was evaluated by positron ... ...

Abstract	Epithelial membrane protein-2 (EMP2) is upregulated in a number of tumors and therefore remains a promising target for monoclonal antibody (mAb)-based therapy. In the current study, image guided therapy for an anti-EMP2 mAb was evaluated by positron emission tomography (PET) in both syngeneic and immunodeficient cancer models expressing different levels of EMP2 in order to enable a better understanding of its tumor uptake and off target accumulation and clearance. The therapeutic efficacy of the anti-EMP2 mAb was initially evaluated in high- and low-expressing tumors, and the mAb reduced tumor load for the high EMP2 expressing 4T1 and HEC-1-A tumors. To create an imaging agent, the anti-EMP2 mAb was conjugated to p-SCN-Bn-deferoxamine (DFO) and radiolabeled with 89Zr. Tumor targeting and tissue biodistribution were evaluated in syngeneic tumor models (4T1, CT26, and Panc02) and human tumor xenograft models (Ramos, HEC-1-A, and U87MG/EMP2). PET imaging revealed radioactive accumulation in EMP2 positive tumors within 24h post-injection, and the signal was retained for 5 days. High specific uptake was observed in tumors with high EMP2 expression (4T1, CT26, HEC-1-A, U87MG/EMP2), with less accumulation in tumors with low EMP2 expression (Panc02, Ramos). Biodistribution at 5 days post-injection revealed that the tumor uptake ranged from 2 to ~16 %ID/cc. The results show that anti-EMP2 mAbs exhibit EMP2-dependent tumor uptake with low off-target accumulation in preclinical cancer models. The development of improved anti-EMP2 antibody fragments may be useful to track EMP2 positive tumors for subsequent therapeutic interventions.
Language	English
Publishing date	2024-02-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	2063563-1
ISSN	1538-8514 ; 1535-7163
ISSN (online)	1538-8514
ISSN	1535-7163
DOI	10.1158/1535-7163.MCT-23-0465
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Article: Heart Uptake of [

Li, Junfeng / Hu, Weidong / Peng, Jiangling / Wong, Patty / Kandeel, Fouad / Olafsen, Tove / Shively, John E

Pharmaceuticals (Basel, Switzerland)

2022 Volume 15, Issue 12

Abstract: The world-wide high incidence of non-alcoholic fatty liver disease (NAFLD) is of concern for its progression to insulin resistance, steatohepatitis and cardiovascular disease (CVD). The increased uptake of fatty acids in critical organs plays a major ... ...

Abstract	The world-wide high incidence of non-alcoholic fatty liver disease (NAFLD) is of concern for its progression to insulin resistance, steatohepatitis and cardiovascular disease (CVD). The increased uptake of fatty acids in critical organs plays a major role in NAFLD progression. Male Ceacam1−/− mice that develop NAFLD, insulin resistance and CVD on normal chow are a potential model for studying the dysregulation of fatty acid uptake. [18F]fluoro-4-thia-oleate ([18F]FTO) was chosen as a fatty acid reporter because of its higher uptake and retention in the heart in an animal model of CVD. Male wild-type (WT) or Ceacam1−/− mice fasted 4−6 h were administered [18F]FTO i.v., and dynamic PET scans were conducted in an MR/PET small animal imaging system along with terminal tissue biodistributions. Quantitative heart image analysis revealed significantly higher uptake at 35 min in Ceacam1−/− (6.0 ± 1.0% ID/cc) vs. WT (3.9 ± 0.6% ID/cc) mice (p = 0.006). Ex vivo heart uptake/retention (% ID/organ) was 2.82 ± 0.45 for Ceacam1−/− mice vs. 1.66 ± 0.45 for WT mice (p < 0.01). Higher kidney and pancreas uptake/retention in Ceacam1−/− was also evident, and the excretion of [18F]FTO into the duodenum was observed for both WT and Ceacam1−/− mice starting at 10 min. This study suggests that the administration of [18F]FTO as a marker of fatty acid uptake and retention may be an important tool in analyzing the effect of NAFLD on lipid dysregulation in the heart.
Language	English
Publishing date	2022-12-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2193542-7
ISSN	1424-8247
ISSN	1424-8247
DOI	10.3390/ph15121577
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Article ; Online: Multimodality PET and Near-Infrared Fluorescence Intraoperative Imaging of CEA-Positive Colorectal Cancer.

Lwin, Thinzar M / Minnix, Megan / Li, Lin / Sherman, Anakim / Hong, Teresa / Wong, Jeffery Y C / Olafsen, Tove / Poku, Erasmus / Bouvet, Michael / Fong, Yuman / Shively, John E / Yazaki, Paul J

Molecular imaging and biology

2023 Volume 25, Issue 4, Page(s) 727–734

Abstract: Purpose: Molecular imaging is a major diagnostic component for cancer management, enabling detection, staging of disease, targeting therapy, and monitoring the therapeutic response. The coordination of multimodality imaging techniques further enhances ... ...

Abstract	Purpose: Molecular imaging is a major diagnostic component for cancer management, enabling detection, staging of disease, targeting therapy, and monitoring the therapeutic response. The coordination of multimodality imaging techniques further enhances tumor localization. The development of a single agent for real-time non-invasive targeted positron emission tomography (PET) imaging and fluorescence guided surgery (FGS) will provide the next generation tool in the surgical management of cancer. Procedures: The humanized anti-CEA M5A-IR800 "sidewinder" (M5A-IR800-SW) antibody-dye conjugate was designed with a NIR 800 nm dye incorporated into a PEGylated linker and conjugated with the metal chelate p-SCN-Bn-deferoxamine (DFO) for zirconium-89 PET imaging ( Results: The Conclusions: This study demonstrates the potential of a pegylated anti-CEA M5A-IR800-Sidewinder for NIR fluorescence/PET/MR multimodality imaging for intraoperative fluorescence guided surgery.
MeSH term(s)	Humans ; Mice ; Animals ; Tissue Distribution ; Positron-Emission Tomography/methods ; Immunoconjugates ; Disease Models, Animal ; Cell Line, Tumor ; Zirconium ; Colorectal Neoplasms/diagnostic imaging ; Colorectal Neoplasms/pathology ; Polyethylene Glycols
Chemical Substances	Immunoconjugates ; Zirconium (C6V6S92N3C) ; Polyethylene Glycols (3WJQ0SDW1A)
Language	English
Publishing date	2023-06-21
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2079160-4
ISSN	1860-2002 ; 1536-1632
ISSN (online)	1860-2002
ISSN	1536-1632
DOI	10.1007/s11307-023-01831-8
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Article: Biodistribution of Intra-Arterial and Intravenous Delivery of Human Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles in a Rat Model to Guide Delivery Strategies for Diabetes Therapies.

Li, Junfeng / Komatsu, Hirotake / Poku, Erasmus K / Olafsen, Tove / Huang, Kelly X / Huang, Lina A / Chea, Junie / Bowles, Nicole / Chang, Betty / Rawson, Jeffrey / Peng, Jiangling / Wu, Anna M / Shively, John E / Kandeel, Fouad R

Pharmaceuticals (Basel, Switzerland)

2022 Volume 15, Issue 5

Abstract: Umbilical cord mesenchymal stem cell-derived extracellular vesicles (UC-MSC-EVs) have become an emerging strategy for treating various autoimmune and metabolic disorders, particularly diabetes. Delivery of UC-MSC-EVs is essential to ensure optimal ... ...

Abstract	Umbilical cord mesenchymal stem cell-derived extracellular vesicles (UC-MSC-EVs) have become an emerging strategy for treating various autoimmune and metabolic disorders, particularly diabetes. Delivery of UC-MSC-EVs is essential to ensure optimal efficacy of UC-MSC-EVs. To develop safe and superior EVs-based delivery strategies, we explored nuclear techniques including positron emission tomography (PET) to evaluate the delivery of UC-MSC-EVs in vivo. In this study, human UC-MSC-EVs were first successfully tagged with I-124 to permit PET determination. Intravenous (I.V.) and intra-arterial (I.A.) administration routes of [
Language	English
Publishing date	2022-05-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2193542-7
ISSN	1424-8247
ISSN	1424-8247
DOI	10.3390/ph15050595
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Article ; Online: Safety and Biodistribution Profile of Poly(styrenyl acetal trehalose) and Its Granulocyte Colony Stimulating Factor Conjugate.

Ko, Jeong Hoon / Forsythe, Neil L / Gelb, Madeline B / Messina, Kathryn M M / Lau, Uland Y / Bhattacharya, Arvind / Olafsen, Tove / Lee, Jason T / Kelly, Kathleen A / Maynard, Heather D

Biomacromolecules

2022 Volume 23, Issue 8, Page(s) 3383–3395

Abstract: Poly(styrenyl acetal trehalose) (pSAT), composed of trehalose side chains linked to a polystyrene backbone via acetals, stabilizes a variety of proteins and enzymes against fluctuations in temperature. A promising application of pSAT is conjugation of ... ...

Abstract	Poly(styrenyl acetal trehalose) (pSAT), composed of trehalose side chains linked to a polystyrene backbone via acetals, stabilizes a variety of proteins and enzymes against fluctuations in temperature. A promising application of pSAT is conjugation of the polymer to therapeutic proteins to reduce renal clearance. To explore this possibility, the safety of the polymer was first studied. Investigation of acute toxicity of pSAT in mice showed that there were no adverse effects of the polymer at a high (10 mg/kg) concentration. The immune response (antipolymer antibody and cytokine production) in mice was also studied. No significant antipolymer IgG was detected for pSAT, and only a transient and low level of IgM was elicited. pSAT was also safe in terms of cytokine response. The polymer was then conjugated to a granulocyte colony stimulating factor (GCSF), a therapeutic protein that is approved by the Federal Drug Administration, in order to study the biodistribution of a pSAT conjugate. A site-selective, two-step synthesis approach was developed for efficient conjugate preparation for the biodistribution study resulting in 90% conjugation efficiency. The organ distribution of GCSF-pSAT was measured by positron emission tomography and compared to controls GCSF and GCSF-poly(ethylene glycol), which confirmed that the trehalose polymer conjugate improved the in vivo half-life of the protein by reducing renal clearance. These findings suggest that trehalose styrenyl polymers are promising for use in therapeutic protein-polymer conjugates for reduced renal clearance of the biomolecule.
MeSH term(s)	Acetals ; Animals ; Granulocyte Colony-Stimulating Factor ; Mice ; Polymers/chemistry ; Proteins/chemistry ; Tissue Distribution ; Trehalose/chemistry
Chemical Substances	Acetals ; Polymers ; Proteins ; Granulocyte Colony-Stimulating Factor (143011-72-7) ; Trehalose (B8WCK70T7I)
Language	English
Publishing date	2022-06-29
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	1526-4602
ISSN (online)	1526-4602
DOI	10.1021/acs.biomac.2c00511
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: PET imaging and treatment of pancreatic cancer peritoneal carcinomatosis after subcutaneous intratumoral administration of a novel oncolytic virus, CF33-hNIS-antiPDL1.

Zhang, Zhifang / Yang, Annie / Chaurasiya, Shyambabu / Park, Anthony K / Kim, Sang-In / Lu, Jianming / Olafsen, Tove / Warner, Susanne G / Fong, Yuman / Woo, Yanghee

Molecular therapy oncolytics

2021 Volume 24, Page(s) 331–339

Abstract: Peritoneal carcinomatosis of gastrointestinal malignancies remains fatal. CF33-hNIS-antiPDL1, a chimeric orthopoxvirus expressing the human sodium iodide symporter (hNIS) and anti-human programmed death-ligand 1 antibody, has demonstrated robust ... ...

Abstract	Peritoneal carcinomatosis of gastrointestinal malignancies remains fatal. CF33-hNIS-antiPDL1, a chimeric orthopoxvirus expressing the human sodium iodide symporter (hNIS) and anti-human programmed death-ligand 1 antibody, has demonstrated robust preclinical activity against pancreatic adenocarcinoma (PDAC). We investigated the ability of CF33-hNIS-antiPDL1 to infect, help detect, and kill peritoneal tumors following intratumoral (i.t.) injection of subcutaneous (s.c.) tumors
Language	English
Publishing date	2021-12-31
Publishing country	United States
Document type	Journal Article
ISSN	2372-7705
ISSN	2372-7705
DOI	10.1016/j.omto.2021.12.022
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Article: Safety and Biodistribution Profile of Poly(styrenyl acetal trehalose) and Its Granulocyte Colony Stimulating Factor Conjugate

Ko, Jeong Hoon / Forsythe, Neil L. / Gelb, Madeline B. / Messina, Kathryn M. M. / Lau, Uland Y. / Bhattacharya, Arvind / Olafsen, Tove / Lee, Jason T. / Kelly, Kathleen A. / Maynard, Heather D.

Biomacromolecules. 2022 June 29, v. 23, no. 8

2022

Abstract	Poly(styrenyl acetal trehalose) (pSAT), composed of trehalose side chains linked to a polystyrene backbone via acetals, stabilizes a variety of proteins and enzymes against fluctuations in temperature. A promising application of pSAT is conjugation of the polymer to therapeutic proteins to reduce renal clearance. To explore this possibility, the safety of the polymer was first studied. Investigation of acute toxicity of pSAT in mice showed that there were no adverse effects of the polymer at a high (10 mg/kg) concentration. The immune response (antipolymer antibody and cytokine production) in mice was also studied. No significant antipolymer IgG was detected for pSAT, and only a transient and low level of IgM was elicited. pSAT was also safe in terms of cytokine response. The polymer was then conjugated to a granulocyte colony stimulating factor (GCSF), a therapeutic protein that is approved by the Federal Drug Administration, in order to study the biodistribution of a pSAT conjugate. A site-selective, two-step synthesis approach was developed for efficient conjugate preparation for the biodistribution study resulting in 90% conjugation efficiency. The organ distribution of GCSF–pSAT was measured by positron emission tomography and compared to controls GCSF and GCSF–poly(ethylene glycol), which confirmed that the trehalose polymer conjugate improved the in vivo half-life of the protein by reducing renal clearance. These findings suggest that trehalose styrenyl polymers are promising for use in therapeutic protein–polymer conjugates for reduced renal clearance of the biomolecule.
Keywords	acute toxicity ; antibodies ; half life ; immune response ; polystyrenes ; positron-emission tomography ; renal clearance ; temperature ; therapeutics ; trehalose
Language	English
Dates of publication	2022-0629
Size	p. 3383-3395.
Publishing place	American Chemical Society
Document type	Article
ISSN	1526-4602
DOI	10.1021/acs.biomac.2c00511
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Antibody vectors for imaging.

Olafsen, Tove / Wu, Anna M

Seminars in nuclear medicine

2010 Volume 40, Issue 3, Page(s) 167–181

Abstract: Noninvasive molecular imaging approaches include nuclear, optical, magnetic resonance imaging, computed tomography, ultrasound, and photoacoustic imaging, which require accumulation of a signal delivered by a probe at the target site. Monoclonal ... ...

Abstract	Noninvasive molecular imaging approaches include nuclear, optical, magnetic resonance imaging, computed tomography, ultrasound, and photoacoustic imaging, which require accumulation of a signal delivered by a probe at the target site. Monoclonal antibodies are high affinity molecules that can be used for specific, high signal delivery to cell surface molecules. However, their long circulation time in blood makes them unsuitable as imaging probes. Efforts to improve antibodies pharmacokinetics without compromising affinity and specificity have been made through protein engineering. Antibody variants that differ in antigen binding sites and size have been generated and evaluated as imaging probes to target tissues of interest. Fast clearing fragments, such as single-chain variable fragment (scFv; 25 kDa), with 1 antigen-binding site (monovalent) demonstrated low accumulation in tumors because of the low exposure time to the target. Using scFv as building block to produce larger, bivalent fragments, such as scFv dimers (diabodies, 50 kDa) and scFv-fusion proteins (80 kDa minibodies and 105 kDa scFv-Fc), resulted in higher tumor accumulation because of their longer residence time in blood. Imaging studies with these fragments after radiolabeling have demonstrated excellent, high-contrast images in gamma cameras and positron emission tomography scanners. Several studies have also investigated antibody fragments conjugated to fluorescence (near infrared dyes), bioluminescence (luciferases), and quantum dots for optical imaging and iron oxides nanoparticles for magnetic resonance imaging. However, these studies indicate that there are several factors that influence successful targeting and imaging. These include stability of the antibody fragment, the labeling chemistry (direct or indirect), whether critical residues are modified, the number of antigen expressed on the cell, and whether the target has a rapid recycling rate or internalizes upon binding. The preclinical data presented are compelling, and it is evident that antibody-based molecular imaging tracers will play an important future role in the diagnosis and management of cancer and other diseases.
MeSH term(s)	Antibodies, Monoclonal/chemistry ; Humans ; Image Enhancement/methods ; Isotope Labeling/trends ; Molecular Probe Techniques/trends ; Radioimmunodetection/trends ; Radiopharmaceuticals/chemistry ; Tomography, Emission-Computed/trends
Chemical Substances	Antibodies, Monoclonal ; Radiopharmaceuticals
Language	English
Publishing date	2010-03-27
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	120248-0
ISSN	1558-4623 ; 0001-2998
ISSN (online)	1558-4623
ISSN	0001-2998
DOI	10.1053/j.semnuclmed.2009.12.005
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Article ; Online: Biodistribution of Intra-Arterial and Intravenous Delivery of Human Umbilical Cord Mesenchymal Stem Cell-Derived Extracellular Vesicles in a Rat Model to Guide Delivery Strategies for Diabetes Therapies

Junfeng Li / Hirotake Komatsu / Erasmus K. Poku / Tove Olafsen / Kelly X. Huang / Lina A. Huang / Junie Chea / Nicole Bowles / Betty Chang / Jeffrey Rawson / Jiangling Peng / Anna M. Wu / John E. Shively / Fouad R. Kandeel

Pharmaceuticals, Vol 15, Iss 595, p

2022 Volume 595

Abstract	Umbilical cord mesenchymal stem cell-derived extracellular vesicles (UC-MSC-EVs) have become an emerging strategy for treating various autoimmune and metabolic disorders, particularly diabetes. Delivery of UC-MSC-EVs is essential to ensure optimal efficacy of UC-MSC-EVs. To develop safe and superior EVs-based delivery strategies, we explored nuclear techniques including positron emission tomography (PET) to evaluate the delivery of UC-MSC-EVs in vivo. In this study, human UC-MSC-EVs were first successfully tagged with I-124 to permit PET determination. Intravenous (I.V.) and intra-arterial (I.A.) administration routes of [ 124 I]I-UC-MSC-EVs were compared and evaluated by in vivo PET-CT imaging and ex vivo biodistribution in a non-diabetic Lewis (LEW) rat model. For I.A. administration, [ 124 I]I-UC-MSC-EVs were directly infused into the pancreatic parenchyma via the celiac artery. PET imaging revealed that the predominant uptake occurred in the liver for both injection routes, and further imaging characterized clearance patterns of [ 124 I]I-UC-MSC-EVs. For biodistribution, the uptake (%ID/gram) in the spleen was significantly higher for I.V. administration compared to I.A. administration (1.95 ± 0.03 and 0.43 ± 0.07, respectively). Importantly, the pancreas displayed similar uptake levels between the two modalities (0.20 ± 0.06 for I.V. and 0.24 ± 0.03 for I.A.). Therefore, our initial data revealed that both routes had similar delivery efficiency for [ 124 I]I-UC-MSC-EVs except in the spleen and liver, considering that higher spleen uptake could enhance immunomodulatory application of UC-MSC-EVs. These findings could guide the development of safe and efficacious delivery strategies for UC-MSC-EVs in diabetes therapies, in which a minimally invasive I.V. approach would serve as a better delivery strategy. Further confirmation studies are ongoing.
Keywords	extracellular vesicles (EVs) ; umbilical cord mesenchymal stem cell (UCMSC) ; diabetes ; I-124 ; positron emission tomography (PET) ; intravenous (I.V.) administration ; Medicine ; R ; Pharmacy and materia medica ; RS1-441
Subject code	571
Language	English
Publishing date	2022-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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