PMID- 34909150 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220105 IS - 2041-6520 (Print) IS - 2041-6539 (Electronic) IS - 2041-6520 (Linking) VI - 12 IP - 45 DP - 2021 Nov 24 TI - Correlative 3D cryo X-ray imaging reveals intracellular location and effect of designed antifibrotic protein-nanomaterial hybrids. PG - 15090-15103 LID - 10.1039/d1sc04183e [doi] AB - Revealing the intracellular location of novel therapeutic agents is paramount for the understanding of their effect at the cell ultrastructure level. Here, we apply a novel correlative cryo 3D imaging approach to determine the intracellular fate of a designed protein-nanomaterial hybrid with antifibrotic properties that shows great promise in mitigating myocardial fibrosis. Cryo 3D structured illumination microscopy (cryo-3D-SIM) pinpoints the location and cryo soft X-ray tomography (cryo-SXT) reveals the ultrastructural environment and subcellular localization of this nanomaterial with spatial correlation accuracy down to 70 nm in whole cells. This novel high resolution 3D cryo correlative approach unambiguously locates the nanomaterial after overnight treatment within multivesicular bodies which have been associated with endosomal trafficking events by confocal microscopy. Moreover, this approach allows assessing the cellular response towards the treatment by evaluating the morphological changes induced. This is especially relevant for the future usage of nanoformulations in clinical practices. This correlative super-resolution and X-ray imaging strategy joins high specificity, by the use of fluorescence, with high spatial resolution at 30 nm (half pitch) provided by cryo-SXT in whole cells, without the need of staining or fixation, and can be of particular benefit to locate specific molecules in the native cellular environment in bio-nanomedicine. CI - This journal is (c) The Royal Society of Chemistry. FAU - Groen, J AU - Groen J AUID- ORCID: 0000-0002-9395-8108 AD - MISTRAL Beamline, Experiments Division, ALBA Synchrotron Light Source Cerdanyola del Valles 08290 Barcelona Spain epereiro@cells.es. FAU - Palanca, A AU - Palanca A AUID- ORCID: 0000-0001-7274-7021 AD - Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria, CSIC 39011 Santander Spain. AD - Department of Anatomy and Cell Biology, University of Cantabria 39011 Santander Spain. FAU - Aires, A AU - Aires A AD - Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA) Paseo de Miramon 194 20014 Donostia San Sebastian Spain alcortajarena@cicbiomagune.es. FAU - Conesa, J J AU - Conesa JJ AD - MISTRAL Beamline, Experiments Division, ALBA Synchrotron Light Source Cerdanyola del Valles 08290 Barcelona Spain epereiro@cells.es. AD - National Center for Biotechnology CSIC (CNB-CSIC), Department of Macromolecular Structures Cantoblanco 28049 Madrid Spain. FAU - Maestro, D AU - Maestro D AD - Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria, CSIC 39011 Santander Spain. FAU - Rehbein, S AU - Rehbein S AD - Helmholtz-Zentrum Berlin fur Materialien und Energie, Bessy II D-12489 Berlin Germany. FAU - Harkiolaki, M AU - Harkiolaki M AD - Beamline B24, Diamond Light Source, Harwell Science and Innovation Campus Didcot Oxfordshire OX11 0DE UK. FAU - Villar, A V AU - Villar AV AUID- ORCID: 0000-0003-4588-9598 AD - Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria, CSIC 39011 Santander Spain. AD - Department of Physiology and Pharmacology, University of Cantabria Avd. Herrera Oria s/n Santander Spain. FAU - Cortajarena, A L AU - Cortajarena AL AUID- ORCID: 0000-0002-5331-114X AD - Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA) Paseo de Miramon 194 20014 Donostia San Sebastian Spain alcortajarena@cicbiomagune.es. AD - Ikerbasque, Basque Foundation for Science 48009 Bilbao Spain. FAU - Pereiro, E AU - Pereiro E AUID- ORCID: 0000-0001-7626-5935 AD - MISTRAL Beamline, Experiments Division, ALBA Synchrotron Light Source Cerdanyola del Valles 08290 Barcelona Spain epereiro@cells.es. LA - eng PT - Journal Article DEP - 20211019 PL - England TA - Chem Sci JT - Chemical science JID - 101545951 PMC - PMC8612387 COIS- There are no conflicts to declare. EDAT- 2021/12/16 06:00 MHDA- 2021/12/16 06:01 PMCR- 2021/10/19 CRDT- 2021/12/15 12:28 PHST- 2021/07/30 00:00 [received] PHST- 2021/10/17 00:00 [accepted] PHST- 2021/12/15 12:28 [entrez] PHST- 2021/12/16 06:00 [pubmed] PHST- 2021/12/16 06:01 [medline] PHST- 2021/10/19 00:00 [pmc-release] AID - d1sc04183e [pii] AID - 10.1039/d1sc04183e [doi] PST - epublish SO - Chem Sci. 2021 Oct 19;12(45):15090-15103. doi: 10.1039/d1sc04183e. eCollection 2021 Nov 24.