{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T14:06:22Z","timestamp":1790777182939,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2022,11,9]],"date-time":"2022-11-09T00:00:00Z","timestamp":1667952000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Deanship of Scientific Research at King Saud University","award":["IFKSURG-346"],"award-info":[{"award-number":["IFKSURG-346"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Due to their interesting size-dependent magnetic characteristics and relative biocompatibility, magnetic superparamagnetic iron oxide (SPIO) nanoparticles have been widely exploited as probes for cell and subcellular structure identification, as well as medication and gene delivery. A thorough understanding of the mechanics of the interaction between nanoparticles and macrophages is vital in managing dynamic processes in nanomedicine. In this study, the interaction behavior and uptake of SPIO nanoparticles by M1- and M2-type macrophages were investigated. Mice monocytes were differentiated into M1 and M2 macrophages, and the uptake of SPIO nanoparticles was studied using a TEM microscope. A high resolution image of 1 nm resolution, an image processing technique, was developed to extract the SPIO-NPs from tomographic TEM microscopic images. Lysosomes appear to be the zones of high concentrations of SPIO inside macrophages. Lysosomes were first selected in each image, and then segmentation by the Otsu thresholding method was used to extract the SPIO-NPs. The Otsu threshold method is a global thresholding technique used to automatically differentiate SPIOs from the background. The SPIO-NPs appear in red colors, and the other pixels in the image are considered background. Then, an estimation of the SPIO-NP uptakes by lysosomes is produced. Higher uptake of all-sized nanoparticles was observed in M1- and M2-type macrophages. An accurate estimation of the number of SPIO-NPs was obtained. This result will help in controlling targeted drug delivery and assessing the safety impact of the use of SPIO-NPs in nanomedicine for humans.<\/jats:p>","DOI":"10.3390\/ijms232213801","type":"journal-article","created":{"date-parts":[[2022,11,10]],"date-time":"2022-11-10T01:48:08Z","timestamp":1668044888000},"page":"13801","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Estimation of SPIO Nanoparticles Uptakes by Macrophages Using Transmission Electron Microscopy"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3638-7916","authenticated-orcid":false,"given":"Adham","family":"Aleid","sequence":"first","affiliation":[{"name":"Department of Biomedical Technology, King Saud University, Riyadh 11433, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5274-084X","authenticated-orcid":false,"given":"Khalid","family":"Alhussaini","sequence":"additional","affiliation":[{"name":"Department of Biomedical Technology, King Saud University, Riyadh 11433, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed","family":"Almijalli","sequence":"additional","affiliation":[{"name":"Department of Biomedical Technology, King Saud University, Riyadh 11433, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1448-4072","authenticated-orcid":false,"given":"Ali S.","family":"Saad","sequence":"additional","affiliation":[{"name":"Department of Biomedical Technology, King Saud University, Riyadh 11433, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1166\/jno.2020.2730","article-title":"Extraction of Iron Oxide Nanoparticles from 3 Dimensional MRI Images Using K-Mean Algorithm","volume":"15","author":"Alanazi","year":"2020","journal-title":"J. Nanoelectron. Optoelectron."},{"key":"ref_2","first-page":"666","article-title":"Visualization and quantification of SPIO nanoparticles in intracellular spaces of macrophages for nanomedicine applications","volume":"27","author":"Saad","year":"2016","journal-title":"Biomed. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1998","DOI":"10.3389\/fimmu.2019.01998","article-title":"Nanoparticles Targeting Macrophages as Potential Clinical Therapeutic Agents Against Cancer and Inflammation","volume":"10","author":"Hu","year":"2019","journal-title":"Front. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2665","DOI":"10.1021\/nn300060u","article-title":"Nanomagnetic sensing of blood plasma protein interactions with iron oxide nanoparticles: Impact on macrophage uptake","volume":"6","author":"Lartigue","year":"2012","journal-title":"ACS Nano"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"7216","DOI":"10.1021\/acsnano.0c02289","article-title":"Polyglycerol grafting shields nanoparticles from protein corona formation to avoid macrophage uptake","volume":"14","author":"Zou","year":"2020","journal-title":"ACS Nano"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e29528","DOI":"10.4161\/tisb.29528","article-title":"Barriers to drug delivery in solid tumors","volume":"2","author":"Sriraman","year":"2014","journal-title":"Tissue Barriers"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"8174","DOI":"10.1039\/C5CS00499C","article-title":"Understanding and Exploiting Nanoparticles\u2019 Intimacy with the Blood Vessel and Blood","volume":"44","author":"Magdiel","year":"2015","journal-title":"Chem. Soc. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5936","DOI":"10.1002\/adfm.201401664","article-title":"Back to basics: Exploiting the innate physico-chemical characteristics of nanomaterials for biomedical applications","volume":"24","author":"Tay","year":"2014","journal-title":"Adv. Funct. Mater."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.biomaterials.2015.01.072","article-title":"Rapid determination of plasmonic nanoparticle agglomeration status in blood","volume":"51","author":"Samir","year":"2015","journal-title":"Biomaterials"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1021\/nl4032549","article-title":"Nanoparticles strengthen intracellular tension and retard cellular migration","volume":"14","author":"Chor","year":"2014","journal-title":"Nano Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1080\/17460441.2020.1733525","article-title":"Targeting macrophages: A novel avenue for cancer drug discovery","volume":"15","author":"Kumar","year":"2020","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1278","DOI":"10.2174\/092986709787846604","article-title":"Iron oxide nanoparticle platform for biomedical applications","volume":"16","author":"Xie","year":"2009","journal-title":"Curr. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"21266","DOI":"10.3390\/ijms141121266","article-title":"Magnetic nanoparticles: Surface effects and properties related to biomedicine applications","volume":"14","author":"Issa","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Almijalli, M., Saad, A., Alhussaini, K., Aleid, A., and Alwasel, A. (2021). Towards Drug Delivery Control Using Iron Oxide Nanoparticles in Three-Dimensional Magnetic Resonance Imaging. Nanomaterials, 11.","DOI":"10.3390\/nano11081876"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Beduneau, A., Ma, Z., Grotepas, C.B., Kabanov, A., Rabinow, B.E., Gong, N., Mosley, R.L., Dou, H., Boska, M.D., and Gendelman, H.E. (2009). Facilitated monocyte-macrophage uptake and tissue distribution of superparmagnetic iron-oxide nanoparticles. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004343"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1038\/nri2448","article-title":"Exploring the full spectrum of macrophage activation","volume":"8","author":"Mosser","year":"2008","journal-title":"Nat. Rev. Immunol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.imbio.2006.06.002","article-title":"Classical and alternative activation of mononuclear phagocytes: Picking the best of both worlds for tumor promotion","volume":"211","author":"Movahedi","year":"2006","journal-title":"Immunobiology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1007\/978-1-59745-396-7_12","article-title":"Derivation and characterization of murine alternatively activated (M2) macrophages","volume":"531","author":"Ho","year":"2009","journal-title":"Methods Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1038\/nmat2442","article-title":"Understanding biophysicochemical interactions at the nanobio interface","volume":"8","author":"Nel","year":"2009","journal-title":"Nat. Mater."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2873","DOI":"10.1007\/s00018-009-0053-z","article-title":"Nanocarriers\u2019 entry into the cell: Relevance to drug delivery","volume":"66","author":"Hillaireau","year":"2009","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_21","first-page":"3284","article-title":"Targeting macrophages using nanoparticles: A potential therapeutic strategy for atherosclerosis","volume":"9","author":"Zang","year":"2021","journal-title":"J. Mater. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"218","DOI":"10.3389\/fimmu.2020.00218","article-title":"Targeting of hepatic macrophages by therapeutic nanoparticle","volume":"11","author":"Colino","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Hufnagel, M., Neuberger, R., Wall, J., Link, M., Friesen, A., and Hartwig, A. (2021). Impact of Differentiated Macrophage-Like Cells on the Transcriptional Toxicity Profile of CuO Nanoparticles in Co-Cultured Lung Epithelial Cells. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22095044"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"19737","DOI":"10.1073\/pnas.1917424117","article-title":"Specific targeting of ovarian tumor-associated macrophages by large, anionic nanoparticles","volume":"117","author":"Haber","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2721","DOI":"10.2217\/nnm-2017-0248","article-title":"Targeted macrophages delivery of rifampicin-loaded lipid nanoparticles to improve tuberculosis treatment","volume":"12","author":"Vieira","year":"2017","journal-title":"Nanomedicine"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"114238","DOI":"10.1016\/j.addr.2022.114238","article-title":"Why nanoparticles prefer liver macrophage cell uptake in vivo","volume":"185","author":"Ngo","year":"2022","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Kang, S.H., Shin, Y.S., Lee, D.-H., Park, I.S., Kim, S.K., Ryu, D., Park, Y., Byun, S.-H., Choi, J.-H., and Hong, S.J. (2022). Interactions of Nanoparticles with Macrophages and Feasibility of Drug Delivery for Asthma. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031622"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Swartzwelter, B.J., Verde, A., Rehak, L., Madej, M., Puntes, V.F., De Luca, A.C., Boraschi, D., and Italiani, P. (2021). Interaction between Macrophages and Nanoparticles: In Vitro 3D Cultures for the Realistic Assessment of Inflammatory Activation and Modulation of Innate Memory. Nanomaterials, 11.","DOI":"10.3390\/nano11010207"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1126\/sciadv.aay1601","article-title":"Nanoparticle interactions with immune cells dominate tumor retention and induce T cell\u2013mediated tumor suppression in models of breast cancer","volume":"6","author":"Korangath","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1002\/cmmi.1516","article-title":"Real-time high-resolution magnetic resonance tracking of macrophage subpopulations in a murine inflammation model: A pilot study with a commercially available cryogenic probe","volume":"8","author":"Luciani","year":"2013","journal-title":"Contrast Media Mol. Imaging"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4487","DOI":"10.1016\/j.bmc.2017.06.040","article-title":"Nanoparticle-macrophage interactions: A balance between clearance and cell-specific targeting","volume":"25","author":"Rattan","year":"2017","journal-title":"Bioorg. Med. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1172\/JCI29950","article-title":"Ly-6chi monocytes dominate hypercholesterolemia-associated monocytosis and give rise to macrophages in atheromata","volume":"117","author":"Swirski","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_33","first-page":"1","article-title":"Preferential magnetic nanoparticle uptake by bone marrow derived macrophages sub populations: Effect of surface coating on polarization, toxicity, and in vivo MRI detection","volume":"15","year":"2013","journal-title":"J. Nanopart. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1007\/s12650-014-0259-5","article-title":"3D Visualization of iron oxide nanoparticles in MRI of inflammatory model","volume":"18","author":"Saad","year":"2014","journal-title":"J. Vis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1564","DOI":"10.1002\/mrm.22843","article-title":"Enhanced magnetic cell labeling efficiency using-NH2 coated MPIOs","volume":"65","author":"Tang","year":"2011","journal-title":"Magn. Reason. Med."},{"key":"ref_36","first-page":"953","article-title":"Enhanced cellular uptake of aminosilane-coated superparamagnetic iron oxide nanoparticles in mammalian cell lines","volume":"7","author":"Zhu","year":"2012","journal-title":"Int. J. Nanomed."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1007\/s11671-008-9226-1","article-title":"Effects of proteins from culture medium on surface property of silanes functionalized magnetic nanoparticles","volume":"4","author":"Chen","year":"2008","journal-title":"Nanoscale Res. Lett."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.colsurfb.2013.02.027","article-title":"On the role of surface composition and curvature on biointerface formation and colloidal stability of nanoparticles in a protein-rich model system","volume":"108","author":"Natte","year":"2013","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1109\/TNANO.2021.3116185","article-title":"Using Otsu\u2019s Method for Image Segmentation to Determine the Particle Density, Surface Coverage and Cluster Size Distribution of 3 nm Si Nanoparticles","volume":"20","author":"Ashraf","year":"2021","journal-title":"IEEE Trans. Nanotechnol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/13801\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:13:22Z","timestamp":1760145202000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/22\/13801"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,9]]},"references-count":39,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["ijms232213801"],"URL":"https:\/\/doi.org\/10.3390\/ijms232213801","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,9]]}}}