Xenogeneic Transplantation Promoted Human Exosome Sequestration in Rat Specific Organs
Purpose: Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model. Methods: Exos were isolated from normal Human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 breast cancer cells. Diameter size and...
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| Published in | Advanced pharmaceutical bulletin Vol. 14; no. 2; pp. 426 - 433 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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Iran
Tabriz University of Medical Sciences
01.07.2024
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| Abstract | Purpose: Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model. Methods: Exos were isolated from normal Human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 breast cancer cells. Diameter size and zeta potential distribution were studied using dynamic light scattering (DLS). The morphology of isolated Exos was monitored by scanning electron microscopy (SEM) images. Using western blotting, protein levels of exosomal tetraspanins were detected. For the in vivo study, Dil-labeled normal and cancer Exos were injected into the tail vein (100 µg exosomal protein/rat) three times at 1-hour intervals. After 24 hours, rats were euthanized and the cellular uptake of Exos was monitored in different organs using immunofluorescence staining (IF). Results: The size distribution and mean zeta potential of HUVEC and MDA-MB-231 cells Exos were 80±29.94 and 64.77±25.49 nm, and −7.58 and −11.8 mV, respectively. Western blotting revealed CD9, CD81, and CD63 in normal and cancer Exos. The SEM images exhibited typical nano-sized round-shape Exo particles. IF staining indicated sequestration of administrated Exos in splenic tissue and lungs. The distribution of Exo in kidneys, aorta, and hepatic tissue was less. These features were more evident in the group that received cancer Exos. We found no obvious adverse effects in rats that received normal or cancer Exos. Conclusion: Normal and cancerous xenogeneic human Exos can be sequestrated prominently in splenic tissue and lungs. Novel delivery approaches and engineering tools are helpful in the target delivery of administrated Exos to the injured sites. |
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| AbstractList | Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model.
Exos were isolated from normal Human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 breast cancer cells. Diameter size and zeta potential distribution were studied using dynamic light scattering (DLS). The morphology of isolated Exos was monitored by scanning electron microscopy (SEM) images. Using western blotting, protein levels of exosomal tetraspanins were detected. For the
study, Dil-labeled normal and cancer Exos were injected into the tail vein (100 µg exosomal protein/rat) three times at 1-hour intervals. After 24 hours, rats were euthanized and the cellular uptake of Exos was monitored in different organs using immunofluorescence staining (IF).
The size distribution and mean zeta potential of HUVEC and MDA-MB-231 cells Exos were 80±29.94 and 64.77±25.49 nm, and -7.58 and -11.8 mV, respectively. Western blotting revealed CD9, CD81, and CD63 in normal and cancer Exos. The SEM images exhibited typical nano-sized round-shape Exo particles. IF staining indicated sequestration of administrated Exos in splenic tissue and lungs. The distribution of Exo in kidneys, aorta, and hepatic tissue was less. These features were more evident in the group that received cancer Exos. We found no obvious adverse effects in rats that received normal or cancer Exos.
Normal and cancerous xenogeneic human Exos can be sequestrated prominently in splenic tissue and lungs. Novel delivery approaches and engineering tools are helpful in the target delivery of administrated Exos to the injured sites. Purpose: Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model. Methods: Exos were isolated from normal Human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 breast cancer cells. Diameter size and zeta potential distribution were studied using dynamic light scattering (DLS). The morphology of isolated Exos was monitored by scanning electron microscopy (SEM) images. Using western blotting, protein levels of exosomal tetraspanins were detected. For the in vivo study, Dil-labeled normal and cancer Exos were injected into the tail vein (100 µg exosomal protein/rat) three times at 1-hour intervals. After 24 hours, rats were euthanized and the cellular uptake of Exos was monitored in different organs using immunofluorescence staining (IF). Results: The size distribution and mean zeta potential of HUVEC and MDA-MB-231 cells Exos were 80±29.94 and 64.77±25.49 nm, and −7.58 and −11.8 mV, respectively. Western blotting revealed CD9, CD81, and CD63 in normal and cancer Exos. The SEM images exhibited typical nano-sized round-shape Exo particles. IF staining indicated sequestration of administrated Exos in splenic tissue and lungs. The distribution of Exo in kidneys, aorta, and hepatic tissue was less. These features were more evident in the group that received cancer Exos. We found no obvious adverse effects in rats that received normal or cancer Exos. Conclusion: Normal and cancerous xenogeneic human Exos can be sequestrated prominently in splenic tissue and lungs. Novel delivery approaches and engineering tools are helpful in the target delivery of administrated Exos to the injured sites. In addition to the direct membrane fusion, the direct interaction of Exo surface ligands with cell surface receptors can also help the uptake procedure.6 8 In recent years, whole-cell- and EV (Exos)-based therapies have been used along with conventional modalities for the treatment of various complications and abnormalities.9 Compared to the whole cell therapy, Exos can in part, but not completely, circumvent the problems associated with crossing biological interfaces and allo-recognition rejection.4 The field of Exo-based drug delivery is at the center of attention for increasing targeting efficiency.10 For this purpose, researchers have purified Exos from diverse biofluids with healthy and cancerous origins, and culture media using different protocols. The injection of allogeneic and xenogeneic Exos can stimulate antigen-presenting cells and allo-/xeno-reactive responses.11 The physicochemical properties and high-rate in vivo biodistribution can increase the likelihood of elimination by hepatic and splenic macrophages.12 It should not be forgotten that the low levels of recognition elements on circulating Exo surface and rapid cell entry can reduce the direct interaction of Exos with immune cells compared to allogeneic/xenogeneic cells.12 In modalities associated with Exo therapy, the delivery of active compounds to the injured site is the subject of debate. Materials and Methods Animal ethics For experimental procedures, permission was obtained from the Local Committee of the Ethics at Tabriz University of Medical Sciences (IR.TBZMED.VCR.REC.1400.350).14 In this study, 15 male Wistar rats, ranging from 6 to 8 weeks old and weighing about 120 g, were used. Morphological assessment The morphology of normal and cancer Exos was investigated using scanning electron microscopy (SEM) microscopes as previously described.16,17 For ТЕМ imaging, one drop (approximately 20 pL) of both purified Exos suspended in PBS was separately placed on carboncoated 300-mesh copper grids and subjected to uranyl acetate staining (2% wt./v). |
| Author | Mahdipour, Mahdi Ghaffari-Nasab, Arshad Mobarak, Halimeh Rahbarghazi, Reza |
| AuthorAffiliation | 4 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 1 Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 2 Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran 3 Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran |
| AuthorAffiliation_xml | – name: 2 Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran – name: 3 Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran – name: 4 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran – name: 1 Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran |
| Author_xml | – sequence: 1 givenname: Halimeh orcidid: 0000-0001-6834-1483 surname: Mobarak fullname: Mobarak, Halimeh organization: Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran., Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran – sequence: 2 givenname: Mahdi orcidid: 0000-0002-2729-4593 surname: Mahdipour fullname: Mahdipour, Mahdi organization: Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran., Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran – sequence: 3 givenname: Arshad surname: Ghaffari-Nasab fullname: Ghaffari-Nasab, Arshad organization: Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran – sequence: 4 givenname: Reza orcidid: 0000-0003-3864-9166 surname: Rahbarghazi fullname: Rahbarghazi, Reza organization: Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran |
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| Cites_doi | 10.3402/jev.v4.27066 10.3402/jev.v4.26238 10.1136/thx.2010.138438 10.1021/acs.molpharmaceut.8b00901 10.1016/j.canlet.2015.10.020 10.1038/s41580-022-00460-3 10.1007/978-0-387-75847-3_8 10.1182/blood-2004-03-0824 10.1016/j.bbcan.2019.04.004 10.1186/s13287-022-03201-7 10.1038/s41598-020-57497-7 10.3892/mmr.2013.1738 10.1186/s12943-015-0426-x 10.1186/s12943-019-0964-8 10.3389/fcell.2021.628103 10.1007/s13770-021-00361-0 10.2147/ijn.s269069 10.3389/fphar.2016.00533 10.1113/jp282053 10.1016/j.ynpai.2022.100095 10.1002/biof.1497 10.1016/j.bioadv.2022.213276 10.17226/12910 10.1186/s13287-021-02596-z 10.1186/s12964-019-0390-y 10.1038/nature15756 10.1016/j.ebiom.2019.03.011 10.7150/thno.51571 10.1002/jcb.22733 10.1186/s40659-020-0273-0 10.1002/advs.201901779 10.1186/s13287-020-01921-2 10.3390/cells11131989 10.1002/0471143030.cb0322s30 10.3389/fimmu.2020.591065 10.1080/20013078.2018.1555419 10.1186/s13578-021-00650-0 10.1073/pnas.1304266110 |
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| Keywords | Human exosomes Cellular uptake Intravenous administration Rat Off-target sequestration |
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| Snippet | Purpose: Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model.... Here, we aimed to study the distribution pattern of normal and cancer xenogeneic exosomes (Exos) and possible interspecies reactions in a rat model. Exos were... In addition to the direct membrane fusion, the direct interaction of Exo surface ligands with cell surface receptors can also help the uptake procedure.6 8 In... |
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| SubjectTerms | Biodistribution Biotechnology Breast cancer Cells cellular uptake human exosomes intravenous administration Medical research Membranes Morphology off-target sequestration Original Proteins rat Scanning electron microscopy |
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| Title | Xenogeneic Transplantation Promoted Human Exosome Sequestration in Rat Specific Organs |
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