NEWS

evFOUNDRY researcher presented a “turbo talk” at Bio Med Eng 21

Ric Gillams (Southampton group) presented a “turbo talk” entitled “Extracellular vesicle isolation in a microfluidic channel” at the Biomedical Engineering meeting, 6-7th September 2021, University of Sheffield, UK.

evFOUNDRY M35 partners’ meeting held in Brescia and telematic

The 6 th and 7 th of July evFOUNDRY members met both in person and telematics for the month 35 partners meeting.

After more than a year, part of the evFOUNDRY consortium (CSGI and LUND) met in person at the University of Brescia for two collaborative days of discussion while all the other members joined in telematic mode.

evFOUNDRY researchers contributed to demonstrate that careful SARS-CoV-2 inactivation practices needs be adopted prior serum Extracellular Vesicle (EV) analysis. 

This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EV analysis.  The Extracellular Vesicles (EVs) contained in the serum have been analyzed upon the application of the main practices used to inactivate SARS-CoV-2, the group of CSGI-Bologna in particular have applied the protocol recently developed to measured the changes induced in the EVs size and nano mechanical properties.

https://ec.europa.eu/info/funding-tenders/opportunities/portal/screen/opportunities/horizon-results-platform/34270

The front cover of Nanoscale Horizons features a new assay developed by evFOUNDRY researchers: A “plasmon-based nanoruler” to measure the mechanical properties of Extracellular Vesicles

The CSGI team of evFOUNDRY developed a conceptual and methodological advancement which allows accessing the stiffness of extracellular vesicles, or more generally of membrane-enveloped nanosized objects, such as viruses. The measurement of this nanoscale property was achieved using a probe of a similar length scale, that is 10-nm gold nanoparticles which behave. Leveraging the plasmonic properties of these sensitive and fast nanoprobes, they set-up a very simple approach, with a straightforward colorimetric readout, easily accessible with basic lab equipment and expertise.   Their discovery has been published and featured on the front cover of Nanoscale Horizons, which is a leading journal for the publication of urgent short reports of exceptionally high quality and innovative nanotechnology.     These findings pave the way for a new field of investigation, where the stiffness of membranous nanoparticles can be related to biological properties, such as pharmacokinetics and cell internalization.   The “plasmon-based stiffness nanoruler” is a reproducible, sensitive, high-throughput, and readily accessible method, which makes it possible the measurement of tiny amounts of sample.

Read more at: https://pubs.rsc.org/en/content/articlelanding/2021/nh/d1nh00012h#!divAbstract

This new research has been warmly welcomed by the colloids community and will be presented shortly at the international conferences of:

- 2021 ACS FALL NATIONAL MEETING, Atlanta, USA, 22-26 August 2021: https://www.agrodiv.org/event/2021-acs-fall-national-meeting/  -  Oral contribution “Probing the mechanical properties of synthetic and biogenic nanosized lipid vesicles with a plasmon based nanoruler” by Debora Berti.

- ECIS2021, Athens, Greece, 5-10 Sep 2021 (https://www.ecis2021.org/)  -  Keynote contribution “Assessing the stiffness of synthetic and natural nanovesicles by a “plasmon-based nanoruler”” by Lucrezia Caselli.

- 13^th EBSA Congress, Vienna, Austria, 24-28 July (http://ebsa.org/portal/node/332)  -  Poster contribution “The response of biogenic lipid vesicles to deformations: how pressure and membrane elasticity affect stiffness” by Andrea Ridolfi

New insights into the nanomechanics of lamellar and non-lamellar membranes

New fundamental contribution to the mechanical properties of different lipid phases have been achieved by researchers within the evFOUNDRY consortium. Two preprints are now available on bioRχiv.

The first one (https://www.biorxiv.org/content/10.1101/2021.06.17.448831v1 deals with the mechanical response of lipid vesicles and Supported Lipid Bilayers (SLBs) to indentation and helps understanding how membrane rigidity and internal pressure contribute to the general response of a vesicle to deformations.

The second one https://www.biorxiv.org/content/10.1101/2021.03.29.437497v1 presents the first mechanical characterization of cubic membranes at the nanoscale and reveals an important relationship between the mechanical response of these architectures and their topology.

evFOUNDRY researcher won the  “Best Interpretative Image” award at ISEV 2021

Andrea Ridolfi contributed to the 10th Annual Meeting ISEV 2021, giving an oral presentation entitled: "AFM-based high throughput mechanical characterization of single EVs from different natural sources". He also won the “Best Interpretative Image” award of the “Image competition" with the image "EV night by Atomic Force Microscopy". 

evFOUNDRY at the International Society for Extracellular Vesicles (ISEV2021) annual meeting

evFOUNDRY contributed to the 10th edition of the International Society for Extracellular Vesicles annual meeting (ISEV2021) https://na.eventscloud.com/website/16865/ with a massive participation. Researchers contributed with:

1. Two oral presentations

a) "Set-up of multi-parametric analytical assays for quantification, quality control and surrogate potency testing of helminth EVs" (Francesca Loria, co-authored by Hansabiomed Life Sciences, University of Aarhus, CSGI-University of Brescia)

b) "AFM-based high throughput mechanical characterization of single EVs from different natural sources" (Andrea Ridolfi, authored by CNR-ISMN, CSGI-University of Florence, CSGI-University of Brescia, University of Aarhus, University of Utrecht, Hansabiomed Life Sciences);

2. One oral poster:

"Pre-processing of bovine milk to clear casein micelles can influence the colloidal and functional properties of milk extracellular vesicles" (from evFOUNDRY project, PhD. Martijn van Herwijnen, co-authored by University of Utrecht, CNR-ISMN, CSGI-University of Florence, CSGI-University of Brescia).

This year, our partner Annalisa Radeghieri chaired the “hot-topic” oral sessions titled “Bone Repair”.

We are proud to announce that Andrea Ridolfi from CNR-ISMN, CSGI-University of Florence won the “Best Interpretative Image” award of the “Image competition. See related news for more details.

evFOUNDRY researchers from Utrecht University published a paper in the Journal of Nutrition

A new publication by the Wauben group from Utrecht University on the characterization of bovine milk extracellular vesicles was published in the Journal of Nutrition! The authors show that milk EVs from pasteurized or Ultra Heat Treated (UHT) milk have altered characteristics compared to milk EVs from raw cows milk.

Journal of Nutrition

The functional characterization of human milk EVs reveals a role in the infants oral mucosa.

In the latest publication by our evFOUNDRY members from Utrecht, the authors show that extracellular vesicles from human milk support the oral cavity epithelial barrier and modulate both innate and adaptive immune responses. This work was published in the prestigious Journal of Extracellular Vesicles. Additionally, Martijn van Herwijnen and Marca Wauben were invited to the present their work in the EV-Journal Club of the International Society of Extracellular Vesicles (ISEV).

This open assess paper can be accessed via: Journal of Extracellular Vesicles

The recording of the EV-Journal Club can be found on YouTube: EV-Journal Club

The Aarhus group members of evFOUNDRY held a 3 day long PhD course ‘Extracellular Vesicles (EVs) – an introduction’

The Aarhus group members of evFOUNDRY held a 3 day long PhD course ‘Extracellular Vesicles (EVs) – an introduction’ at Aarhus University, Denmark, 16-18 March 2021. Due to the pandemic, the course was held online. The participants learned the basics of EVs, experimental considerations and functional and translational aspects of the EVs. The course was structured with lectures, group projects, and online practical work.

WORKSHOP on massive production of EVs

On 28^th-29^th October 2021 at the University of Brescia (Brescia, Italy) and virtually, it will take place the WORKSHOP on massive production of EVs Co-organized with the FETopen projects evFOUNDRY and VES4US.  

Application & Abstract submission opens April 15^th 2021 and closes June 1^st 2021. This workshop will be free for accepted applicants and limited to 50 attendees.    

www.isev.org/events

evFOUNDRY M29 Telematic Partners’ Meeting

On February 1st and 2nd  2021 the evFOUNDRY team met in telematic mode to review the latest 3 months of activity and plan next moves.  The meeting saw the participation of all the project researchers in two days of intense discussion. It helped to cheer up a bit from the pandemic difficulties and restrictions. The meeting began with the welcome to the new Project Officer Athina Zampara, who met “in person” the evFOUNDRY team for the first time. 

Biogenic, Hybrid and Synthetic Vesicles

evFOUNDRY researchers Paolo Bergese and Costanza Montis co-edited, together with Kimberly Hamad Schifferili (UMASS - US) a special issue in Biochimica et Biophysica Acta (BBA) - General Subjects on “Biogenic, Hybrid and Synthetic Vesicles”. The full issue is available at the following link:

https://www.sciencedirect.com/journal/biochimica-et-biophysica-acta-bba-general-subjects/vol/1865/issue/4

Check it out! 

evFOUNDRY researcher present at the ISEV Infectious Diseases Meeting 2021

At the International Society of Extracellular Vesicles’ Infectious Diseases Meeting 2021 on the 25th-28th of January, Anne Borup of the Aarhus-group presented a 10-min talk entitled “Comparison of three enrichment methods for immunomodulatory extracellular vesicles from helminths”. The talk highlighted the findings of the evFOUNDRY work regarding the helminth EVs.

evFOUNDRY M27 Telematic Technical Meeting

On 23rd November 2020 the evFOUNDRY team met in telematic mode to review the 3 months of activity and plan next moves. The meeting saw the participation of researchers from each partner evFOUNDRY consortium (in particular all the of the young scientists hired on the grant were present) involved in one day of intense discussion (see picture)

evFOUNDRY researchers published a paper in Micromachines.

By combining DLD with electric fields (eDLD), Prof. Tegenfeldt's group show separation of a variety of nano and micro-sized particles primarily by their zeta potential – which is correlated to their surface charge.

The separation of nanoscale liposomes of different lipid compositions was shown as a proof of principle of this method, with strong relevance for biomedicine and for EVs separation.

Ongoing work focuses on using this type of device to separate EVs based on size and surface charge.

The paper has been labelled as "Feature Paper", "Editor's Choice", and is currently at the top of the "Highly Accessed Articles" list of the journal."

https://www.mdpi.com/2072-666X/11/11/1014

Repository link: Zenodo

On November 2020 Patrizia Andreozzi joined evFOUNDRY team (CSGI-Florence)

Patrizia Andreozzi recently joined the evFOUNDRY team in Florence, Italy. She builded up her strong background on Physical Chemistry of Colloid Systems, in particular in synthesis and characterization of nanomaterials beginning her carrier with a master thesis and PhD in Chemistry at The Sapienza University, Rome. She worked as Post-Doc at Department of Chemical Sciences and Technologies of University of Rome Tor Vergata, in Rome (Italy), at National Nanotechnology Laboratory (NNL) - Nanochemistry Division, Istituto di Nanoscienze del CNR, Lecce (Italy) and at IRCCS Carlo Besta Neurological Institute Fundation, IFOM-Fondazione Istituto FIRC di Oncologia Molecolare, Department of Nanomedicine, in Milan (Italy). From 2016 to 2019 she was Researcher at Centro de Investigación Cooperativa en Biomateriales - CIC biomaGUNE, Department of Soft Matter Nanotecnology, in San Sebastian (Spain).

Finally on November 2020 she joined as Post-doc the evFOUNDRY Team: Welcome on board Patrizia!

Martijn van Herwijnen speaker for SNEV

Martijn van Herwijnen (Utrecht University) gave a virtual talk entitled “Solving the milk EV puzzle” for the Student Network on EVs (SNEV). SNEV is a virtual society run by students who are interested in the EV field. Its mission is to connect EV research students internationally, to promote discussion, share knowledge, network, and establish collaboration with other students and early career researchers on the extracellular vesicles field through series of virtual events.

https://snevresearch.wordpress.com/blog/

“Extracellular vesicles in regenerative medicine”

Researchers from CSGI in collaboration with Dr. Anna C. Berardi from Unit of Immunohematology and Transfusion Medicine, Spirito Santo Hospital, Pescara, Italy, published a chapter entitled “Extracellular vesicles in regenerative medicine” in the book “Nanomaterials for Theranostics and Tissue Engineering: Techniques, Trends and Applications”. Micro and Nano Technologies series, edited by Filippo Rossi and Alberto Rainer. The chapter gives a concise introduction of the two leading techniques in regenerative medicine: cell-based therapies and tissue engineering. It provides an overview of the most promising non-EV cell-free therapies and discuss EV biological and physicochemical properties and then cover their potential role in regenerative medicine according to the state-of-the-art literature.

ISBN: 978-0-12-817838-6

http://hdl.handle.net/11379/535723

https://doi.org/10.1016/B978-0-12-817838-6.00002-4

evFOUNDRY researchers awarded for the “Best Flash Presentation” at μTAS 2020

Prof. Tegenfeldt’s group (Lund University, Sweden) won the “Best Flash Presentation” award at the 24th International Conference on Miniaturized Systems for Chemistry and Life Sciences (µTAS 2020; 4-9 October 2020). The 1-minute video has attracted a large crowd of audience to the 1-hour live zoom room where Bao Ho presented the poster entitled “High Throughput Extracellular Vesicle Sorting Using Electrokinetic Deterministic Lateral Displacement” (see picture).

Check the video out, you won’t regret it!!!

https://www.youtube.com/watch?v=D6L3oTlj5fg&feature=youtu.be&t=33

The evFOUNDRY consortium develops an AFM-based method for the high-throughput nanomechanical and morphological measurement of individual EVs

evFOUNDRY researchers recently published a paper in which they describe a novel Atomic Force Microscopy (AFM)-based methodology for the simultaneous nanomechanical and morphological analysis of several hundred individual nanosized EVs within the hour time scale. This methodology only requires basic AFM instrumentation and skills for its experimental part, and freely available software for data analysis. In the paper, authors demonstrate how their AFM-based method can be used to quickly detect EV samples contamination by non vesicular aggregates, and to quickly attest the presence of EVs even in samples for which no established assays and/or commercial kits are available, thus making it a valuable tool for the rapid assessment of EV samples during the development of isolation/enrichment protocols by EV researchers.

https://iris.unibs.it/handle/11379/534530

https://www.biorxiv.org/content/10.1101/854539v1

https://pubs.acs.org/doi/abs/10.1021/acs.analchem.9b05716

evFOUNDRY says on the future

evFOUNDRY will participate by its coordinator to the session entitled "Voices from the future: Pathfinder stories, people, visions", organized on Sept. 22nd, 09:45 - 12:30 CETA in the frame of the Future Tech Week. The session will be opened by a keynote speech from the neuroscientist Nobel Laureate Professor Edvard I. Moser and followed by a roundtable on the topic of extracellular vesicles involving the coordinators of the cluster of the seven EU-funded ongoing FET projects on extracellular vesicles. Participation is telematic and free, follow the link below to register.

http://www.fetfx.eu/?na=view&id=27

https://cordis.europa.eu/article/id/411728-the-evmanifesto-released

https://youtu.be/EuOYNe3CieY

evFOUNDRY intercepted by the “Innovation Radar

evFOUNDRY has been selected by Innovation Radar as a key innovator project thanks to the “Microfluidic device for EVs extraction” and “Scalable production and SOPs for quality standard nanosized EVs” actions. Innovation Radar is a European Commission initiative to identify high potential innovations and innovators in EU-funded research. It supports projects by suggesting a range of targeted actions to assist them in fulfilling their potential in the market. Both evFOUNDRY actions obtained the following scores:     Market maturity: Business Ready     Market creation potential: High     Innovation Topic: Excellent Science.

More info at: https://www.innoradar.eu/resultbykeyword/evfoundry

evFOUNDRY research selected for an Outstanding Oral Abstract Award at the ISEV2020 Virtual Meeting

On Wednesday 22 July 2020, Andrea Zendrini has been recognized for an Outstanding Oral Abstract Award at the ISEV2020 Virtual Meeting for his abstract The CONAN assay: purity grade and concentration of EV microliter formulations by colloidal nanoplasmonics. His abstract spun off our first collective research paper, which presents key implementations of the COlloidal NANoplasmonic (CONAN) assay developed within the project. It was selected, with other two, among the about 150 oral abstracts featured at the Meeting.
Andrea is a post-doc researcher at Paolo Bergese's group (Università degli Studi di Brescia and CSGI) and at Paolo Ajmone's group (Università Cattolica del Sacro Cuore di Piacenza). 

https://www.isev.org/mpage/2020Home

https://doi.org/10.3389/fbioe.2019.00452

https://iris.unibs.it/handle/11379/528221

Fluorescent Labeling of Helminth Extracellular Vesicles Using an In Vivo Whole Organism Approach

Researchers from Aarhus and Lund University, and in collaboration with Aalborg University, have published a paper in Biomedicines. Here they labelled extracellular vesicles by utilizing parasitic helminths inability to synthesize lipids by culturing helminths with fluorescent lipids. These were taken up by the parasite and subsequently released incorporated into the extracellular vesicles. The released extracellular vesicles were characterized with proteomics and Cryo-TEM and uptake was shown in a human cell line. As the inability of synthesizing lipids is general in parasites, it is believed that this novel methods can be used for tracking extracellular vesicles in the field of parasite-host communication.

https://doi.org/10.3390/biomedicines8070213

https://pure.au.dk/portal/da/publications/fluorescent-labeling-of-helminth-extracellular-vesicles-using-an-in-vivo-whole-organism-approach(6f641b5e-82cd-4feb-8550-dd155d78cc0a).html

evFOUNDRY back on eagle’s wings

evFOUNDRY team enters in COVID-19 lockdown phase 3 with a breath of optimism: Brescia and Aarhus units confirm their collaboration with a student exchange. Research teams of Università degli Studi di Brescia (Department of Molecular and Translational Medicine) and Aarhus University (Department of Clinical Medicine, Infectious Disease), led by Prof. Paolo Bergese and Prof. Peter Nejsum, respectively, will be even more connected thanks to the future biotechnologist Giorgio. Training in Nejsum’s lab will complete his formative internship as well as produce substantial part of the experimental results for his thesis project, being a unique possibility with a strong curricular impact. Nejsum’s lab will also benefit of his visit, as he will bring the expertise and know-how learned during the part of his formative internship already attended in Bergese’s lab.

Gold nanoparticles interacting with synthetic lipid rafts: an AFM investigation

Researchers from CSGI published a paper on Journal of Microscopy. In this work, they explored the interaction of gold nanoparticles (AuNPs) with synthetic lipid rafts, formed on Supported Lipid Bilayers. Atomic Force Microscopy (AFM) and Neutron Reflectivity measurements (performed at the Heinz Maier-Leibnitz Zentrum, MLZ) were used to characterize the lipid bilayers and to image the AuNPs adsorbed on them. The thickness mismatch present along the rafts' boundaries is thought to generate energetically favourable conditions for the adsorption of NPs, but a direct proof of this mechanism was still missing to date. In the manuscript, AFM images clearly show that AuNPs preferentially target raft's edges when adsorbing on the bilayer. 

https://doi.org/10.1111/jmi.12910

https://doi.org/10.1101/2020.05.03.075101

Key Exploitable Result (KER) with relevance to COVID-19 from our Utrecht University partner

In a just released research paper, Marca Wauben and coworkers show the inhibitory effects of human milk EVs on T cell proliferation and cytokine release, as well as the inhibition of TLR-triggering on epithelial cells. This hints to a potential nasal/oral application to dampen overactive immune responses, e.g. during the immunopathological responses in lung (and possibly also in gut) reported during SARS-CoV infection.

https://ec.europa.eu/info/funding-tenders/opportunities/portal/screen/opportunities/horizon-results-platform/23443

https://doi.org/10.1101/2020.04.29.068841

A paper published in Journal of Colloid and Interface Science, on extracellular vesicles-based supported lipid bilayers as 2D platforms to study nano-bio interfaces

Researchers from CSGI (Department of Chemistry, University of Florence, Department of Translational Medicine, University of Brescia, CNR Bologna, Italy) made a cover in the journal
Journal of Colloid and Interface Science, with their study on the interaction between superparamagnetic iron oxide nanoparticles and biogenic supported lipid bilayers (SLBs) formed through vesicle fusion of Extracellular Vesicles.

The experimental results highlight a strong interaction of nanoparticles with biogenic SLBs. This evidence strongly suports the hypotheses on the peculiar structure of biogenic SLBs, with cushioned non-flat areas and extended exposed surface; in addition, it suggests that these features are relevant in the response of biogenic membranes to nano-objects.

·        Cover picture in Journal of Colloid and Interface Science (see picture). It is an Inside Front Cover.

https://flore.unifi.it/handle/2158/1218287#.YDYl05NKhsM

Collective fingerprinting of EV subpopulations by Fourier-transform Infrared (FT-IR) spectroscopy

evFOUNDRY researchers, in collaboration with Stefania Federici and Ivano Alessandri from the Dept. of Mechanical and Industrial Engineering and Dept. of Information Engineering (University of Brescia), published a paper on Journal of Extracellular Vesicles. EV subtype classification is an emerging topic and authors challenged FT-IR spectroscopy to fingerprint EV subpopulations separated using differential centrifugation steps. Large, medium and small EVs from murine prostate cancer (TRAMP-C2) and murine skin melanoma (B16) cell culture media were enriched according to their different sedimentation properties. The combination of FT-IR spectroscopy and spectral exploration with chemometric tools (Principal Component Analysis) were able to fingerprint the EV subpopulations both vertically (deriving from the same cell line) and horizontally (among different cell lines). This work revealed the versatility of this collective approach that can be applied for grading the intrinsic high level of heterogeneity of different EV subtypes.

https://doi.org/10.1080/20013078.2020.1741174

https://iris.unibs.it/handle/11379/529275

A paper published in Journal of Colloid and Interface Science, on the interaction between gold nanoparticles and synthetic liposomes.

The CSGI evFOUNDRY team has joined efforts with a multidisciplinary group of researchers from ESRF (Grenoble, France) and  ILL (Grenoble, France) to investigate the mechanistic details of the interaction of gold nanoparticles, coated by citrate anion, with synthetic liposomes. The results of this fundamental study will appear in Journal of Colloid and Interface Science. 
Thanks to an ensemble of optical, structural and surface techniques, we identified the citrate-lipid ligand exchange at the biomimetic interface as the molecular origin of cooperative behavior, which leads to the formation of clusters of gold nanoparticles on the bilayer. Although this study has a fundamental nature, the implications of membrane-induced clusterization of nanoparticles can be extended to biogenic membranes, as in the case of extracellular vesicles.

https://doi.org/10.1016/j.jcis.2020.03.123

https://iris.unibs.it/handle/11379/529276

Anne Borup, a member of the Aarhus group of evFOUNDRY, has been volunteering at the Aarhus University Hospital during the COVID-19 pandemic

Anne Borup, a member of the Aarhus group of evFOUNDRY, has been volunteering at the Aarhus University Hospital during the COVID-19 pandemic. Here a drive-in Corona-test system has been set up in order to ensure safety, minimal spread and test of people suspected to be infected with the Coronavirus. Of high priority is testing of health care professionals with a critical function so they could return to their much-needed services. Anne volunteered at the call center where they received calls from health care professionals that needed to be tested for Corona and subsequently evaluate and organize them for a test in the drive-in.

Ric Gillams and Hywel Morgan (University of Southampton) developed a Personal Respirator for Healthcare Professionals Treating COVID-19 (PeRSo)

Ric Gillams and Hywel Morgan, evFOUNDRY's researchers from the University of Southampton have developed a prototype personal respirator called ‘PeRSo’, intended to address the limitations of the existing protective equipment doctors and nurses are wearing on the wards. Our prototype consists of a fabric hood which covers the wearer’s head, and a plastic visor to protect their face. A small portable unit delivers clean air through a High Efficiency Particulate Air (HEPA) filter to the wearer from a battery-powered fan pack mounted on a belt. In a rapid development process we have gone from a problem specification to working prototypes within a week, and conducted preliminary tests.

https://www.southampton.ac.uk/publicpolicy/support-for-policymakers/policy-projects/perso.page?fbclid=IwAR0ACv2UC-SEL3rlSvJVYoEJzB6y5vN7rBVa5VymbDC7voOu6b4uPyZ_CPs

Ric Gillams described the PerSo to BBC South Today and Times News. 

https://www.youtube.com/watch?v=t3yVrU19idg&feature=youtu.be

https://www.youtube.com/watch?v=LOwVs8aDxgw&feature=youtu.be

CONAN for evFOUNDRY

evFOUNDRY researchers in collaboration with Andrea Zendrini - to date post-doc researcher at Università degli Studi di Brescia and at Università Cattolica di Piacenza - published a protocol that describes how to assign purity grade and titrate extracellular vesicle solutions of a few microliters. The assay is an augmented version of the COlorimetric NANoplasmonic (CONAN) assay developed in 2015 by Paolo Bergese's group. It has been implemented in the first year of activity of evFOUNDRY, being now part of the standard operating procedures (SOPs) for  preparing quality standard nanosized EVs from bovine milk and Ascaris media.

https://doi.org/10.3389/fbioe.2019.00452

https://iris.unibs.it/handle/11379/528221

The Aarhus group members of evFOUNDRY held a 3 day long Ph.D. course ‘Extracellular Vesicles (EVs) – an introduction’

The Aarhus group members of evFOUNDRY held a 3 day long Ph.D. course ‘Extracellular Vesicles (EVs) – an introduction’ at Aarhus University,
Denmark, March 2020. The participants learned the basics of EVs, experimental considerations and functional and translational aspects of the EVs. The course was structured with lectures, group projects, and laboratory work.

evFOUNDRY M18 meeting in Aarhus (Denmark)

On February 4th-6th 2020, the evFOUNDRY team met in Aarhus (hosted by Aarhus University - https://pure.au.dk/portal/en/persons/peter-nejsum(ee468e32-0a8b-4cd3-9473-60335cf27df9).html) to review the first 18 months of activity and plan next moves. The meeting saw the participation of researchers from each partner of the evFOUNDRY consortium (see picture) involved in three days of intense discussion.

During this special occasion, two ESAB members (Andrew Hill https://www.andyfhill.com/ and Pierre Joseph https://www.laas.fr/public/en/mile) attended an evFOUNDRY meeting for the first time.

On 4thFebruary the Workshop on "knowledge management and IPR" was organized within the collaboration with the team of FET-Briefing project (fetbriefing.eu). The Workshop was organized in a telematic mode, through the direct connection from Aarhus to the live event held in Stuttgart, Germany.

Aarhus University is offering a 3-day PhD course with the title “Extracellular Vesicles – an introduction” in March 2020

Aarhus University is offering a 3-day PhD course with the title “Extracellular Vesicles – an introduction” in March 2020.
 
More details can be found here:

https://service.health.au.dk/modules/Course/mypage/viewfulldesc/courseID/1321

Costanza Montis (evFOUNDRY young researcher) authored with Debora Berti a contribution to celebrate 15th Anniversary of Soft Matter

To celebrate the 15th Anniversary of Soft Matter Costanza Montis (evFOUNDRY young researcher) and Debora Berti have been chosen to write their personal perspectives on what are the hot topics in their area, and where they think their sub-field is going. Their contribution reviews the state of art on hybrid soft matter assemblies composed of inorganic nanoparticles (NP) and lamellar or non-lamellar lipid bilayers and the the latest developments on the use of lipid bilayers as coating agents for inorganic NPs are presented. Costanza Montis finally acknowledges evFOUNDRY Project.

https://pubs.rsc.org/en/journals/articlecollectionlanding?sercode=sm&themeid=7751e5a5-7500-4435-ad72-2d6381abc2dd

https://flore.unifi.it/handle/2158/1175074.2#.YDYmTpNKhsM

Andrea Ridolfi (evFOUNDRY member, CSGI-Bologna) prensented a poster at the Workshop "Bill2019"

On 11-13 December 2019 Andrea Ridolfi (CSGI-Bologna) partecipated with a poster entitled “Nanomechanical properties of synthetic and natural lipid bilayers through Atomic Force Microscopy (AFM): from Supported Lipid Bilayers (SLBs) to vesicles” to the Workshop “Bill2019” Grenoble, France.

Extracellular vesicles and nano-friends featured in Biomaterials Science back cover

evFOUNDRY researchers of the University of Brescia and CSGI explain on the first 2020 issue of Biomaterials Science how scientists are learning to exploit the wealth of nanoparticles that populate the secretome, such as extracellular vesicles and lipoproteins. The review article has been also featured in the issue back cover.  

http://dx.doi.org/10.1039/c9bm01007f

https://iris.unibs.it/handle/11379/526980

The Southampton group of evFOUNDRY participated in the UK EV forum, hosted by The UK Society for Extracellular Vesicles (UKEV) in London.

On the 10rd of December 2019, the Southampton group of evFOUNDRY participated in the UK EV forum, hosted by The UK Society for Extracellular Vesicles (UKEV) in London, UK. Ric Gillams presented a poster and flash talk showing results related to evFOUNDRY (see picture).

Biological membranes and EVs

Paolo Bergese (evFOUNDRY coordinator) and Marca Wauben (Utrecht University partner) contributed to set the ISEV position paper on biological membranes in EV biogenesis, stability, uptake, and cargo transfer. The paper assesses current knowledge of several aspects which are key to evFOUNDRY, including EV structural and colloidal properties at the individual level and in relation to their interaction with surfaces and nanomaterials.   

https://www.tandfonline.com/doi/full/10.1080/20013078.2019.1684862

https://iris.unibs.it/handle/11379/526637

evMANIFESTO released.

The 1ST CLUSTERING EVENT FOR EXTRACELLULAR VESICLES FET-OPEN PROJECTS, held in Palermo in November 06th 2019, involved the 5 EU funded ongoing FET OPEN projects on EVs, counting participation of 34 research and industrial organizations from 15 EU countries. The first collective action of the newborn cluster was producing the attached position document, the evMANIFESTO, which aims at highlighting the current EU effort in EV research within the FET program and the urgency of introducing and securing more firmly the EV-related theme as a cornerstone in the research agenda in EU. 

You can find the document in the results section:

http://www.evfoundry.eu/results/results.php#res-26

evFOUNDRY sponsored a special session on Extracellular Vesicles (EVs) within the workshop "Science through Scanning Probe Microscopy 2019" StSPM19

evFOUNDRY sponsored a special session on Extracellular Vesicles (EVs) within the workshop "Science through Scanning Probe Microscopy 2019" StSPM19, held at the National Research Council of Bologna on November 21st and 22nd 2019. The session was focused on the study of EVs through Atomic Force Microscopy; among the speakers: Paolo Bergese and Andrea Ridolfi  for evFOUNDRY, Gijs Wuite (VU University, Amsterdam) and Pietro Parisse (Elettra-Synchrotron, Trieste) as external contributions.

http://www.bo.ismn.cnr.it/stspm19ev/

evFOUNDRY at EVIta 1st Symposium in Palermo, Italy.

evFOUNDRY members organized and participated at the 1st symposium of the Italian Society of Extracellular Vesicles held in Palermo from the 6th to the 8th of November 2019. Annalisa Radeghieri (CSGI-Brescia) organized the meeting as part of EVIta directive board. Paolo Bergese (CSGI-Brescia) was selected for oral presentation "Projecting extracellular vesicles in two dimension" as well as Marco Brucale (CSGI-Bologna) "Nanomechanics of individual extracellular vesicles". Lucia Paolini (CSGI-Brescia) presented a poster. Francesco Valle (CSGI-Bologna), Natasa Zarovni (Hansa-Biomed) and Francesca Loria (Hansa-Biomed) participated at the symposium. Peter Nejsum (Aarus University) gave a morning lecture on “A new level of complexity in host-parasite interaction- Extracellular Vesicles“

evFOUNDRY involved in the 1st Clustering Event for EVs FET-OPEN projects in Palermo (Italy)

On 6th of November 2019 in Palermo (Italy) evFOUNDRY was actively involved together with 4 ongoing FET projects: INDEX, VES4US, GLADIATOR and MINDGAP in the 1st Clustering Event for EVs FET-OPEN projects. 34 organizations from 15 countries were present to the event during which the EVs network has taken off. A letter of Intent to support EVs research was signed and an EVs Manifesto will be created.

evFOUNDRY member at 1st Lugano ExoDay: Technical Challenges in Exosome Research

Lucia Paolini was selected for a flash presentation at the 1st Lugano ExoDay on: “EV subpopulations fingerprinting by Fourier-transform Infrared (FT-IR) spectroscopy”.

The workshop was organized the 25th of October by several Swiss research institutes: University of Southern Switzerland, Cardiocentro Ticino Foundation, Ente Ospedaliero Cantonale. 

A paper published in Advanced Materials Technologies, highlighting the potential of Electrokinetic Deterministic Lateral Displacement in sorting sub-micron particles

Researchers from Tegenfeldt group, Lund University, Sweden (Jason Beech, Bao Ho, Jonas Tegenfeldt) and Guiducci group, EPFL, Switzerland (Kevin Keim, Carlotta Guiducci) made a cover in the journal Advanced Materials Technologies with their novel metal-coated Electrokinetic Deterministic Lateral Displacement (Electrokinetic DLD) devices. These devices bring about several advantages: tuneable particle separations, significant reduction the critical size, and increase in the dynamic range with switching times on the order of seconds. Most notably, in a device having gap size of around 10 µm, polystyrene particles as small as 250 nm can be displaced, demonstrating two-orders of magnitude reduction in critical size using Electrokinetic DLD, as compared to conventional DLD.
https://doi.org/10.1002/admt.201900339

https://zenodo.org/record/3352780#.XZHEh1Uzapo

FET seminar on the evFOUNDRY

An evFOUNDRY presentation starred the FET seminar series on Thursday October 10th at the European Commission Covent Garden building (Brussels). The seminar was actively participated by a varied audience, including EU scientific project officers, economists and policy makers.
 

evFOUNDRY first Review Meeting in Brussels

On the 11th of October 2019 all members of evFOUNDRY Consortium met at Research Executive Agency in Brussels for the first Reporting Period Review Meeting.

Our project officer, Adriana Godeanu-Metz, chaired the meeting sessions.

All the activities conducted during in the different work packages have been presented and discussed with the reviewers.

The Aarhus group of evFOUNDRY participated in the 1st annual meeting in The Danish Society of Extracellular Vesicles (DSEV) held in Aarhus, Denmark.

On the 3rd of October 2019, the Aarhus group of evFOUNDRY participated in the 1st annual meeting in The Danish Society of Extracellular Vesicles (DSEV) held in Aarhus, Denmark. Peter Nejsum and Anne Borup are on the board and founders of the society and were responsible for organizing the event. Anne presented a poster of results related to evFOUNDRY (see picture).

evFOUNDRY took part at the MEETmeTONIGHT (European Researchers’ Night) and Future Tech Week events.

On September 27th evFOUNDRY took part at the MEETmeTONIGHT (European Researchers’ Night) and Future Tech Week  events.

CSGI members introduced students, kids and general public to evFOUNDRY aims and strategies with a ludic-scientific stand. Venue of the event has been Santa Giulia museum in Brescia, Italy.

http://www.meetmetonight.it/

http://futuretechweek.fetfx.eu/

Debora Berti, member of the CSGI team has been appointed in the Scientific Advisory Board for SweDeliver

Debora Berti has been appointed as SAB member for Swedish Drug Delivery Center for the period 2020-2024 with a budget of 3M Euro.

https://www.vinnova.se/en/news/2019/06/efforts-on-world-class-research-environments/

evFOUNDRY M12 meeting in Bologna (Italy)

On September 5-6 2019, the evFOUNDRY team met in Bologna (hosted by ISMN – CNR  - www.ismn.cnr.it) to review the first 12 months of activity and plan next moves also in the perspective of the review meeting that will be held in Bruxelles in October. The meeting saw the participation of researchers from each partner of the evFOUNDRY consortium (see picture) involved in two days of intense discussion.

Workshop on CONAN assay in Brescia (Italy)

On the 4th of September, a workshop was held in Brescia, Italy for members of the evFOUNDRY project. Bao Dang Ho (Lund, Sweden) and Francesca Loria (Estonia, Hansa-Biomed) attended a theoretical and practical course on Colorimetric Nanoplasmonic (CONAN) assay developed in Paolo Bergese’s lab.

On the picture, from the left: Lucia Paolini, Bao Dang Ho and Francesca Loria.

evFOUNDRY got to Singapore

On 30 July 2019 Paolo Bergese introduced evFOUNDRY at the National University of Singapore (NUS), while giving a research seminar on "Nanotechnology of Cell Communication". The seminar was hosted by Prof. Rachel Ee (Dep. of Pharmacy, Faculty od Science) and Prof. James Kah (Dep. of Biomedical Engineering, Faculty of Engineering).

http://www.ee-research-group.com/

http://bioeng.nus.edu.sg/nnl/

On Nature Nanotechnology Paolo Bergese comments with other bio-nano experts on transparency and reproducibility in nanomedicine

Joy Wolfram, Sara Busatto, Mauro Ferrari and Paolo Bergese contributed, together with other international bio-nano experts, to comment the paper "Minimum information reporting in bio–nano experimental literature - MIRIBEL" . The correpondence is now published by Nature Nanotechnology. Their piece advocates integrating MIRIBEL (which is focused on synthetic nanoparticles) with specific considerations related to biogenic nanoparticles such as extracellular vesicles.

https://www.nature.com/articles/s41565-019-0496-9

https://www.nature.com/articles/s41565-018-0246-4

evFOUNDRY at XLVII National meeting of the Physical Chemistry Division of the Italian Chemistry Society, Rome 1-4 July 2019

evFoundry members attended the XLVII National meeting of the Physical Chemistry Division of the Italian Chemistry Society (Physics Department, "Sapienza" University of Rome, 1-4 July 2019) with several contributions:

Costanza Montis with a talk entitled "Interaction of inorganic nanoparticles with synthetic and biogenic lipid membranes: fundamental understanding and biomedical applications'"

Andrea Ridolfi with a poster: "Nanomechanical classification of extracellular vesicles" (see figure)

Debora Berti with a talk entitled  "Nanostructured materials interacting with synthetic and natural lipid mesophases: challenges and opportunities"

See the meeting website for more details: http://congressodcf2019.it/english/

Costanza Montis (evFOUNDRY personnel, CSGI) selected for a presentation in Young investigator perspective section at 9th International Colloids Conference

Costanza Montis gave a talk entitled "Interaction of inorganic nanoparticles with synthetic and biogenic lipid membranes: fundamental understanding and biomedical applications" in the Young Investigator Perspective section at the 9th International Colloids Conference, Sitges (Barcellona, Spain), June 16-19 2019. 

See the meeting website for more details: https://www.elsevier.com/events/conferences/international-colloids-conference

Workshop milk EV isolation at Utrecht University

From May 27 to May 29, Utrecht University was the stage for a workshop of milk EV isolation. In the group of Marca Wauben, visitors Richard Gillams (Southampton, UK) and Bao Dang Ho (Lund, Sweden) experienced first-handedly what its like to process raw cow's milk and to prepare size exclusion columns and density gradients needed for further purification of milk EVs.
 
Congratulations Ric and Bao, for your first batch of purified milk EVs!

ISEV Special Achievement Award 2019 - Recipient: Marca Wauben for recognition of her extraordinary service to EV science and to ISEV

The International Society for Extracellular Vesicles (ISEV) is pleased to announce two Special Achievement Awards that were presented at the ISEV2019 annual meeting in Kyoto, Japan, 24–28 April 2019. ISEV Special Achievement Awards are given each year to one or more individuals who have made outstanding contributions to the field of extra- cellular vesicle (EV) research or have performed extraor- dinary service to ISEV. In 2019, ISEV Special Achievement Awards were presented to Professors Takahiro Ochiya, PhD, and Marca Wauben, PhD (see Figure)

https://www.isev.org/page/ISEVSpecialAchievementAward

https://www.tandfonline.com/doi/full/10.1080/20013078.2019.1620080

Workshop for evFOUNDRY-members in Aarhus (DK)

On the 24th of May, a workshop was held in Aarhus, Denmark for members of the evFOUNDRY. It was a full day of discussion and demonstration of how Peter Nejsum’s group isolate and characterize EVs from Ascaris Suum helminths. Richard Gillams (Southhampton, UK) and Bao Dang Ho (Lund, Sweden) participated.

On the picture, from the left Richard Gilliams, Anne Borup and Bao Dang Ho.

evFOUNDRY at ISEV annual meeting 2019 in Kyoto, Japan

evFOUNDRY will be at the annual meeting of the International Society of Extracellular Vesicles (ISEV) (Kyoto from 24th-28th April 2019), contributing to several sessions:

Martijn van Herwijnen (Utrecht University) will give a lecture on “Therapeutic Potential of Milk EVs” at the educational day (24th April)

Annalisa Radeghieri (CSGI) will give an oral presentation entitled “Different ATT isoforms are associated to EVs from ATT type II deficient patients” at the “EVs in Blood and Blood Disorders” session (25th April)

Lucia Paolini and Martijn van Herwijnen will present a poster on “The extracellular vesicle foundry: evFOUNDRY” at the “Network/society” session and will attend the ISEV Board (26th April)

Marca Wauben (Utrecht University, Secretary General of ISEV and part of the Organizing Committee of ISEV2019) will chair the “Plenary Session on RNA” (27th April).

Lucia Paolini will present two posters: “Fourier-transform Infrared Spectroscopy (FT-IR) to fingerprint EV subpopulations as a whole” and “Taxonomy of individual EVs by nanomechanics” at the “Advances in EV Quantification and Characterization” poster session (27th April). 

See the meeting program for more details: https://www.isev.org/page/ISEV2019

Lucia Paolini invited talk at the “Mini-Workshop on Extracellular Vesicles: from biophysical properties to therapeutic applications”

On Thursday April 11 Lucia Paolini (evFOUNDRY personnel, CSGI) gave an invited talk at the “Mini-Workshop on Extracellular Vesicles: from biophysical properties to therapeutic applications” within EXOTHERA project funded by Interreg Italia-Österreich programme. The talk presented “evFOUNDRY: the extracellular vesicle foundry” project (see flayer for details).

Anne Borup (involved in the evFOUNDRY) have recently received two awards

First, she was awarded ‘best poster presentation’ at the PhD day, 2019 at Aarhus University and secondly she was awarded the ‘young scientist award’ for best poster presentation at the Spring Symposium 2019 in Copenhagen hosted by the Danish Society for Parasitology.

evFOUNDRY M7 meeting in Lund

On March 27th-29th 2019, the evFOUNDRY team met in Lund (hosted by LINXS - Lund Institute of Advanced Neutron and X-ray Studies http://www.linxs.lu.se) to review the first 6 months of activity and plan next moves. The meeting saw for the first time the participation of all the involved young researchers (see picture, from the left Karolina Mothander (Sweden), Lucia Paolini (Italy), Richard Gillams (UK), Andrea Ridolfi (Italy), Anne Borup (Denmark), Anders T Boysen (Denmark), Costanza Montis (Italy), Francesca Loria (Estonia), Bao Dang Ho (Sweden), Martijn van Herwijnen (The Netherlands) not in the picture).
 

Interview on milk Extracellular Vesicles by Martijn van Herwijnen (evFOUNDRY personnel)

The faculty of Veterinary Medicine from Utrecht University has released a new volume of VetScience magazine. This special issue on Future Food includes interviews with Professor of Intercellular Communication Marca Wauben and Martijn van Herwijnen, who explains about his work about milk-derived Extracellular Vesicles (in Dutch). http://www.uu.nl/sites/default/files/vetscience_nov2018.pdf

Ph.D. course: Extracellular vesicles (EVs) – an introduction

 Members of the Evfoundry consortium have taken the initiative, and for the first time, been running the 3 day long Ph.D. course 'Extracellular vesicles (EVs) – an introduction' at Aarhus University, Denmark, March 2019. There were 10 participants from different universities in Denmark. The participants were introduced to EVs and different aspects of EV biology and how to characterize and explore the function of EVs. During the laboratory exercises the participants enriched for EVs and characterized their sizes and numbers and finally, used western blots to identify specific markers of EVs.

Innovation hub on extracellular vesicles in Lombardy Region

evFOUNDRY will participate to a kick off event for establishing an innovation hub on extracellular vesicles in Lombardy Region. The meeting will be held on May 9th, 2019 in Milano (see flyer for details). 

Annalisa Radeghieri (evFOUNDRY personnel) receives funding for engineering extracellular vesicles to deliver antitumor agents to triple negative breast cancer organoids

Annalisa Radeghieri (evFOUNDRY personnel) has received 120,000 euro (approximately USD 137,000) in funding in the framework of an Italian Education and Research Ministry (MIUR-PRIN) grant. The project aims at “Development of biotechnological nanoparticle platform for the delivery of antitumor therapies using Patient Derived-Organoids library of Breast cancer”. It brings in collaboration University of Milano-Bicocca, University of Milan, National Research Council and University of Rome Tor Vergata. 

http://www.miur.gov.it/

Professor Peter Nejsum (evFOUNDRY personnel) receives million-funding for development of new treatment method.

Danish Professor Peter Nejsum (evFOUNDRY personnel) has received DKK 3 million (approximately USD 457,800) in funding for a project to develop a new method to diagnose and treat diseases. The aim is to discover new immunotherapies and diagnose cancer through blood tests. But first step on the journey goes to the local abattoir to collect a bucket of roundworm. 

https://medwatch.dk/secure/Top_picks_in_english/article11041255.ece

Processing large volumes of EV sources. First study supported by evFOUNDRY published on Cells.

Sara Busatto and Paolo Bergese (evFOUNDRY PI) co-authored with Joy Wolfram and colleagues from the Mayo Clinic and the Houston Methodist Research Institute a paper on the use of tangential flow filtration (TFF) for highly efficient concentration of nanosized EVs from large volumes of fluid. The study contributes to evFOUNDRY effort in exploring methods for robust massive separation of EVs based on size.
https://doi.org/10.3390/cells7120273

Debora Berti, member of the CSGI team has been appointed in the Editorial Board of Advances in Colloid and Interface Science.

Debora Berti, member of the CSGI team has been appointed in the Editorial Board of Advances in Colloid and Interface Science, an international journal devoted to experimental and theoretical developments in Interfacial and Colloidal Phenomena and their implications in Biology, Chemistry, Physics and Technology. https://www.journals.elsevier.com/advances-in-colloid-and-interface-science/editorial-board

MISEV2018 is online

Researchers involved in evFOUNDRY contributed to the last update of the “Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines”, which was published online in the Journal of Extracellular Vesicles at the beginning of this week. https://www.tandfonline.com/doi/full/10.1080/20013078.2018.1535750

evFOUNDRY researcher Lucia Paolini selected for FET2RIN business training

evFOUNDRY researcher Lucia Paolini was selected for the FET2RIN training course (three workshops from mid-December 2018 to mid-February 2019 in Brussels).
The training, in collaboration with META, European Crowdfunding Network (ECN), and European Business and Innovation Centre Network (EBN), will help to look at FET project from a business perspective, assessing its strengths and weaknesses on the route to commercialization (http://www.fet2rin.com/home).

KICK OFF PARTNERS’ MEETING

KICK OFF PARTNERS’ MEETING On September 12 2018, at Plesso di SANTA TERESA Florence, next to the workshop (https://www.quotidiano.net/blog/malpelo/ricerca-vescicole-extracellulari-come-taxi-per-farmaci-e-vaccini-33.2061 http://www.meetthelifesciences.it/it/archvio/831-nanotecnologie-evfoundry-nuovi-approcci-ed-utilizzi-delle-vescicole-extracellulari) it’s time for the partners meeting.

evFOUNDRY researchers participate in the foundation of EVITA, the Italian Society of Extracellular Vesicles

On October 5 2018, Lucia Paolini (evFOUNDRY recruited researcher) with Annalisa Radeghieri  (evFOUNDRY personnel, CSGI), Natasa Zarovni (evFOUNDRY personnel, HansaBioMed Life Sciences) and Paolo Bergese (evFOUNDRY coordinator) will participate in the foundation of the Italian Society of Extracellular Vesicles - EVITA.  The meeting will be held at CNR headquarters in Rome.  EVITA will connect leading Italian scientists from all the disciplines involved in EV research in the effort to advance EV research nationally and globally, in tight relationship with other EV Societies, including The International Society for Extracellular Vesicles (https://www.isev.org), the Spanish GEIVEX (http://geivex.org) and the American Society for Exosomes and Microvesicles (https://www.asemv.org).  

Annalisa Radeghieri invited talk at the International Conference on Bioengineering and Cell Therapy

On Friday September 21 Annalisa Radeghieri  (evFOUNDRY personnel, CSGI) will give an invited talk at the International Conference on Bioengineering and Cell Therapy.

The talk will be focused on the potentiality of extracellular vesicles in regenerative medicine. 

Kick off workshop

On September 12 2018, at Plesso di SANTA TERESA Florence , evFOUNDRY kicks off and opens to academic and non-academic communities with a free workshop on “Extracellular vesicles, a breakthrough in precision medicine and nanotechnology”, which sees the partners lecturing on their role and science in the project.

PRESS: quotidiano.net

Vescicole extracellulari come navette, consegnano farmaci e vaccini. https://www.quotidiano.net/blog/malpelo/ricerca-vescicole-extracellulari-come-taxi-per-farmaci-e-vaccini-33.2061 .

PRESS: Meet the Life Sciences

Nanotecnologie: nuovi approcci ed utilizzi delle vescicole extracellulari. http://www.meetthelifesciences.it/it/archvio/831-nanotecnologie-evfoundry-nuovi-approcci-ed-utilizzi-delle-vescicole-extracellulari