2024-03-29T00:35:41Zhttp://repository.helmholtz-hzi.de/oai/requestoai:repository.helmholtz-hzi.de:10033/6209182019-08-30T11:27:16Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Fuhrmann, Gregor
author
Neuer, Anna Lena
author
Herrmann, Inge K
author
2017-04-07
Extracellular vesicles (EVs) have gained increasing attention as novel disease biomarkers and as promising therapeutic agents. These cell-derived, phospholipid-based particles are present in many - if not all - physiological fluids. They have been shown to govern several physiological processes, such as cell-cell communication, but also to be involved in pathological conditions, for example tumour progression. In infectious diseases, EVs have been shown to induce host immune responses and to mediate transfer of virulence or resistance factors. Here, we discuss recent developments in using EVs as diagnostic tools for infectious diseases, the development of EV-based vaccines and the use of EVs as potential anti-infective entity. We illustrate how EV-based strategies could open a viable new avenue to tackle current challenges in the field of infections, including barrier penetration and growing resistance to antimicrobials.
Extracellular vesicles - A promising avenue for the detection and treatment of infectious diseases? 2017 Eur J Pharm Biopharm
1873-3441
28396279
10.1016/j.ejpb.2017.04.005
http://hdl.handle.net/10033/620918
European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V
Extracellular vesicles - A promising avenue for the detection and treatment of infectious diseases?
oai:repository.helmholtz-hzi.de:10033/6213522019-08-30T11:33:29Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Fuhrmann, Gregor
author
Chandrawati, Rona
author
Parmar, Paresh A
author
Keane, Timothy J
author
Maynard, Stephanie A
author
Bertazzo, Sergio
author
Stevens, Molly M
author
2018-02-23
Extracellular vesicles (EVs) have recently gained significant attention as important mediators of intercellular communication, potential drug carriers, and disease biomarkers. These natural cell-derived nanoparticles are postulated to be biocompatible, stable under physiological conditions, and to show reduced immunogenicity as compared to other synthetic nanoparticles. Although initial clinical trials are ongoing, the use of EVs for therapeutic applications may be limited due to undesired off-target activity and potential "dilution effects" upon systemic administration which may affect their ability to reach their target tissues. To fully exploit their therapeutic potential, EVs are embedded into implantable biomaterials designed to achieve local delivery of therapeutics taking advantage of enzyme prodrug therapy (EPT). In this first application of EVs for an EPT approach, EVs are used as smart carriers for stabilizing enzymes in a hydrogel for local controlled conversion of benign prodrugs to active antiinflammatory compounds. It is shown that the natural EVs' antiinflammatory potential is comparable or superior to synthetic carriers, in particular upon repeated long-term incubations and in different macrophage models of inflammation. Moreover, density-dependent color scanning electron microscopy imaging of EVs in a hydrogel is presented herein, an impactful tool for further understanding EVs in biological settings.
Engineering Extracellular Vesicles with the Tools of Enzyme Prodrug Therapy. 2018 Adv. Mater. Weinheim
1521-4095
29473230
10.1002/adma.201706616
http://hdl.handle.net/10033/621352
Advanced materials (Deerfield Beach, Fla.)
Engineering Extracellular Vesicles with the Tools of Enzyme Prodrug Therapy.
oai:repository.helmholtz-hzi.de:10033/6214822019-11-20T02:02:10Zcom_10033_620613com_10033_620656col_10033_620614col_10033_620685col_10033_620657
00925njm 22002777a 4500
dc
Frank, Julia
author
Richter, Maximilian
author
de Rossi, Chiara
author
Lehr, Claus-Michael
author
Fuhrmann, Kathrin
author
Fuhrmann, Gregor
author
2018-08-17
Extracellular vesicles (EVs) are natural nanoparticles that play important roles in intercellular communication and are increasingly studied for biosignalling, pathogenesis and therapy. Nevertheless, little is known about optimal conditions for their transfer and storage, and the potential impact on preserving EV-loaded cargoes. We present the first comprehensive stability assessment of different widely available types of EVs during various storage conditions including -80 °C, 4 °C, room temperature, and freeze-drying (lyophilisation). Lyophilisation of EVs would allow easy handling at room temperature and thus significantly enhance their expanded investigation. A model enzyme, β-glucuronidase, was loaded into different types of EVs derived from mesenchymal stem cells, endothelial cells and cancer cells. Using asymmetric flow field-flow fractionation we proved that the model enzyme is indeed stably encapsulated into EVs. When assessing enzyme activity as indicator for EV stability, and in comparison to liposomes, we show that EVs are intrinsically stable during lyophilisation, an effect further enhanced by cryoprotectants. Our findings provide new insight for exploring lyophilisation as a novel storage modality and we create an important basis for standardised and advanced EV applications in biomedical research.
2045-2322
30120298
10.1038/s41598-018-30786-y
http://hdl.handle.net/10033/621482
Extracellular vesicles protect glucuronidase model enzymes during freeze-drying.
oai:repository.helmholtz-hzi.de:10033/6216652019-08-30T11:34:20Zcom_10033_620613com_10033_620618col_10033_620685col_10033_620619
00925njm 22002777a 4500
dc
Schulz, Eilien
author
Goes, Adriely
author
Garcia, Ronald
author
Panter, Fabian
author
Koch, Marcus
author
Müller, Rolf
author
Fuhrmann, Kathrin
author
Fuhrmann, Gregor
author
2018-11-28
Up to 25,000 people die each year from resistant infections in Europe alone, with increasing incidence. It is estimated that a continued rise in bacterial resistance by 2050 would lead up to 10 million annual deaths worldwide, exceeding the incidence of cancer deaths. Although the design of new antibiotics is still one way to tackle the problem, pharmaceutical companies investigate far less into new drugs than 30 years ago. Incorporation of antibiotics into nanoparticle drug carriers ("nanoantibiotics") is currently investigated as a promising strategy to make existing antibiotics regain antimicrobial strength and overcome certain types of microbial drug resistance. Many of these synthetic systems enhance the antimicrobial effect of drugs by protecting antibiotics from degradation and reducing their side effects. Nevertheless, they often cannot selectively target pathogenic bacteria and - due to their synthetic origin - may induce side-effects themselves. In this work, we present the characterisation of naturally derived outer membrane vesicles (OMVs) as biocompatible and inherently antibiotic drug carriers. We isolated OMVs from two representative strains of myxobacteria, Cystobacter velatus Cbv34 and Sorangiineae species strain SBSr073, a bacterial order with the ability of lysing other bacterial strains and currently investigated as sources of new secondary metabolites. We investigated the myxobacterias' inherent antibacterial properties after isolation by differential centrifugation and purification by size-exclusion chromatography. OMVs have an average size range of 145-194 nm. We characterised their morphology by electron cryomicroscopy and found that OMVs are biocompatible with epithelial cells and differentiated macrophages. They showed a low endotoxin activity comparable to those of control samples, indicating a low acute inflammatory potential. In addition, OMVs showed inherent stability under different storage conditions, including 4 °C, -20 °C, -80 °C and freeze-drying. OMV uptake in Gram-negative model bacterium Escherichia coli (E. coli) showed similar to better incorporation than liposome controls, indicating the OMVs may interact with model bacteria via membrane fusion. Bacterial uptake correlated with antimicrobial activity of OMVs as measured by growth inhibition of E. coli. OMVs from Cbv34 inhibited growth of E. coli to a comparable extent as the clinically established antibiotic gentamicin. Liquid-chromatography coupled mass spectrometry analyses revealed the presence of cystobactamids in OMVs, inhibitors of bacterial topoisomerase currently studied to treat different Gram-negative and Gram-positive pathogens. This work, may serve as an important basis for further evaluation of OMVs derived from myxobacteria as novel therapeutic delivery systems against bacterial infections.
1873-4995
30292423
10.1016/j.jconrel.2018.09.030
http://hdl.handle.net/10033/621665
Journal of Controlled Release
Biogenic drug carriers
Electron cryomicroscopy
Extracellular vesicles
Myxobacteria
Nanoantibiotics
Outer membrane vesicles
Biocompatible bacteria-derived vesicles show inherent antimicrobial activity.
oai:repository.helmholtz-hzi.de:10033/6216692019-08-30T11:24:29Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Théry, Clotilde
author
Witwer, Kenneth W
author
Aikawa, Elena
author
Alcaraz, Maria Jose
author
Anderson, Johnathon D
author
Andriantsitohaina, Ramaroson
author
Antoniou, Anna
author
Arab, Tanina
author
Archer, Fabienne
author
Atkin-Smith, Georgia K
author
Ayre, D Craig
author
Bach, Jean-Marie
author
Bachurski, Daniel
author
Baharvand, Hossein
author
Balaj, Leonora
author
Baldacchino, Shawn
author
Bauer, Natalie N
author
Baxter, Amy A
author
Bebawy, Mary
author
Beckham, Carla
author
Bedina Zavec, Apolonija
author
Benmoussa, Abderrahim
author
Berardi, Anna C
author
Bergese, Paolo
author
Bielska, Ewa
author
Blenkiron, Cherie
author
Bobis-Wozowicz, Sylwia
author
Boilard, Eric
author
Boireau, Wilfrid
author
Bongiovanni, Antonella
author
Borràs, Francesc E
author
Bosch, Steffi
author
Boulanger, Chantal M
author
Breakefield, Xandra
author
Breglio, Andrew M
author
Brennan, Meadhbh Á
author
Brigstock, David R
author
Brisson, Alain
author
Broekman, Marike LD
author
Bromberg, Jacqueline F
author
Bryl-Górecka, Paulina
author
Buch, Shilpa
author
Buck, Amy H
author
Burger, Dylan
author
Busatto, Sara
author
Buschmann, Dominik
author
Bussolati, Benedetta
author
Buzás, Edit I
author
Byrd, James Bryan
author
Camussi, Giovanni
author
Carter, David RF
author
Caruso, Sarah
author
Chamley, Lawrence W
author
Chang, Yu-Ting
author
Chen, Chihchen
author
Chen, Shuai
author
Cheng, Lesley
author
Chin, Andrew R
author
Clayton, Aled
author
Clerici, Stefano P
author
Cocks, Alex
author
Cocucci, Emanuele
author
Coffey, Robert J
author
Cordeiro-da-Silva, Anabela
author
Couch, Yvonne
author
Coumans, Frank AW
author
Coyle, Beth
author
Crescitelli, Rossella
author
Criado, Miria Ferreira
author
D’Souza-Schorey, Crislyn
author
Das, Saumya
author
Datta Chaudhuri, Amrita
author
de Candia, Paola
author
De Santana, Eliezer F
author
De Wever, Olivier
author
del Portillo, Hernando A
author
Demaret, Tanguy
author
Deville, Sarah
author
Devitt, Andrew
author
Dhondt, Bert
author
Di Vizio, Dolores
author
Dieterich, Lothar C
author
Dolo, Vincenza
author
Dominguez Rubio, Ana Paula
author
Dominici, Massimo
author
Dourado, Mauricio R
author
Driedonks, Tom AP
author
Duarte, Filipe V
author
Duncan, Heather M
author
Eichenberger, Ramon M
author
Ekström, Karin
author
EL Andaloussi, Samir
author
Elie-Caille, Celine
author
Erdbrügger, Uta
author
Falcón-Pérez, Juan M
author
Fatima, Farah
author
Fish, Jason E
author
Flores-Bellver, Miguel
author
Försönits, András
author
Frelet-Barrand, Annie
author
2001-3078
10.1080/20013078.2018.1535750
http://hdl.handle.net/10033/621669
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
oai:repository.helmholtz-hzi.de:10033/6219872019-10-22T01:38:55Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Labib, Rola M
author
Ayoub, Iriny M
author
Michel, Haidy E
author
Mehanny, Mina
author
Kamil, Verena
author
Hany, Meryl
author
Magdy, Mirette
author
Moataz, Aya
author
Maged, Boula
author
Mohamed, Ahmed
author
2019-01-01
The present study investigates the wound healing potential of three chitosan-based topical preparations loaded with either tea tree essential oil, rosemary essential oil or a mixture of both oils in vivo. Essential oils of M. alternifolia and R. officinalis were analyzed using GC/MS. Essential oil-loaded chitosan topical preparations were formulated. Wound healing potential was evaluated in vivo using an excision wound model in rats. GC/MS analysis of M. alternifolia and R. officinalis essential oils revealed richness in oxygenated monoterpenes, representing 51.06% and 69.61% of the total oil composition, respectively. Topical application of chitosan-based formulation loaded with a mixture of tea tree and rosemary oils resulted in a significant increase in wound contraction percentage compared to either group treated with individual essential oils and the untreated group. Histopathological examination revealed that topical application of tea tree and rosemary oil combination demonstrated complete re-epithelialization associated with activated hair follicles. The high percentage of oxygenated monoterpenes in both essential oils play an important role in the antioxidant and wound healing potential observed herein. Incorporation of tea tree and rosemary essential oils in chitosan-based preparations in appropriate combination could efficiently promote different stages of wound healing.
PLoS One. 2019 Sep 16;14(9):e0219561. doi: 10.1371/journal.pone.0219561. eCollection 2019.
1932-6203
31525200
10.1371/journal.pone.0219561
http://hdl.handle.net/10033/621987
PLOS ONE
Appraisal on the wound healing potential of Melaleuca alternifolia and Rosmarinus officinalis L. essential oil-loaded chitosan topical preparations.
oai:repository.helmholtz-hzi.de:10033/6220392019-12-06T02:03:50Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Woith, Eric
author
Fuhrmann, Gregor
author
Melzig, Matthias F
author
2019-11-14
It is known that extracellular vesicles (EVs) are shed from cells of almost every type of cell or organism, showing their ubiquity in all empires of life. EVs are defined as naturally released particles from cells, delimited by a lipid bilayer, and cannot replicate. These nano- to micrometer scaled spheres shuttle a set of bioactive molecules. EVs are of great interest as vehicles for drug targeting and in fundamental biological research, but in vitro culture of animal cells usually achieves only small yields. The exploration of other biological kingdoms promises comprehensive knowledge on EVs broadening the opportunities for basic understanding and therapeutic use. Thus, plants might be sustainable biofactories producing nontoxic and highly specific nanovectors, whereas bacterial and fungal EVs are promising vaccines for the prevention of infectious diseases. Importantly, EVs from different eukaryotic and prokaryotic kingdoms are involved in many processes including host-pathogen interactions, spreading of resistances, and plant diseases. More extensive knowledge of inter-species and interkingdom regulation could provide advantages for preventing and treating pests and pathogens. In this review, we present a comprehensive overview of EVs derived from eukaryota and prokaryota and we discuss how better understanding of their intercommunication role provides opportunities for both fundamental and applied biology.
Int J Mol Sci. 2019 Nov 14;20(22). pii: ijms20225695. doi: 10.3390/ijms20225695.
1422-0067
31739393
10.3390/ijms20225695
http://hdl.handle.net/10033/622039
International Journal of Molecular Sciences
archaea
cross-kingdom RNAi
eukaryota
extracellular vesicles
interkingdom communication
prokaryota
Extracellular Vesicles-Connecting Kingdoms.
oai:repository.helmholtz-hzi.de:10033/6220602020-01-04T02:02:30Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Schulz, Eilien
author
Karagianni, Anna
author
Koch, Marcus
author
Fuhrmann, Gregor
author
2019-12-02
In recent years, extracellular vesicles (EVs) and outer membrane vesicles (OMVs) have become an extensive and diverse field of research. They hold potential as diagnostic markers, therapeutics and for fundamental biological understanding. Despite ongoing studies, numerous information regarding function, content and stability of EVs remains unclear. If EVs and OMVs ought to be used as therapeutics and in clinical environments, their stability is one of the most important factors to be considered. Especially for formulation development, EVs and OMVs need to be stable at higher temperatures. To the best of our knowledge, very little work has been published regarding heat stability of neither EVs nor OMVs. In the present study, we investigated B lymphoblastoid cell-derived EVs and OMVs derived from myxobacterial species Sorangiineae as model vesicles. We exposed the vesicles to 37 °C, 50 °C, 70 °C and 100 °C for 1 h, 6 h and 24 h, and also autoclaved them. Interestingly, physico-chemical analyses such as size, particle concentration and protein concentration showed minor alterations, particularly at 37 °C. Flow cytometry analysis emphasised these results suggesting that after heat impact, EVs and OMVs were still able to be taken up by macrophage-like dTHP-1 cells. These data indicate that both mammalian and bacterial vesicles show intrinsic stability at physiological temperature. Our findings are important to consider for vesicle formulation and for advanced bioengineering approaches.
Eur J Pharm Biopharm. 2019 Dec 2. pii: S0939-6411(19)31309-8. doi: 10.1016/j.ejpb.2019.11.010.
1873-3441
31805356
10.1016/j.ejpb.2019.11.010
http://hdl.handle.net/10033/622060
European Journal of Pharmaceutics and Biopharmaceutics
autoclaving
drug carriers
extracellular vesicles
flow cytometry
heat stability
lymphoblastoid cells
myxobacteria
outer membrane vesicles
Hot EVs - how temperature affects extracellular vesicles.
oai:repository.helmholtz-hzi.de:10033/6221222020-02-07T03:09:37Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Hoppstädter, Jessica
author
Dembek, Anna
author
Linnenberger, Rebecca
author
Dahlem, Charlotte
author
Barghash, Ahmad
author
Fecher-Trost, Claudia
author
Fuhrmann, Gregor
author
Koch, Marcus
author
Kraegeloh, Annette
author
Huwer, Hanno
author
Kiemer, Alexandra K
author
2019-01-01
Glucocorticoids (GCs) are widely prescribed therapeutics for the treatment of inflammatory diseases, and endogenous GCs play a key role in immune regulation. Toll-like receptors (TLRs) enable innate immune cells, such as macrophages, to recognize a wide variety of microbial ligands, thereby promoting inflammation. The interaction of GCs with macrophages in the immunosuppressive resolution phase upon prolonged TLR activation is widely unknown. Treatment of human alveolar macrophages (AMs) with the synthetic GC dexamethasone (Dex) did not alter the expression of TLRs -1, -4, and -6. In contrast, TLR2 was upregulated in a GC receptor-dependent manner, as shown by Western blot and qPCR. Furthermore, long-term lipopolysaccharide (LPS) exposure mimicking immunosuppression in the resolution phase of inflammation synergistically increased Dex-mediated TLR2 upregulation. Analyses of publicly available datasets suggested that TLR2 is induced during the resolution phase of inflammatory diseases, i.e., under conditions associated with high endogenous GC production. TLR2 induction did not enhance TLR2 signaling, as indicated by reduced cytokine production after treatment with TLR2 ligands in Dex- and/or LPS-primed AMs. Thus, we hypothesized that the upregulated membrane-bound TLR2 might serve as a precursor for soluble TLR2 (sTLR2), known to antagonize TLR2-dependent cell actions. Supernatants of LPS/Dex-primed macrophages contained sTLR2, as demonstrated by Western blot analysis. Activation of metalloproteinases resulted in enhanced sTLR2 shedding. Additionally, we detected full-length TLR2 and assumed that this might be due to the production of TLR2-containing extracellular vesicles (EVs). EVs from macrophage supernatants were isolated by sequential centrifugation. Both untreated and LPS/Dex-treated cells produced vesicles of various sizes and shapes, as shown by cryo-transmission electron microscopy. These vesicles were identified as the source of full-length TLR2 in macrophage supernatants by Western blot and mass spectrometry. Flow cytometric analysis indicated that TLR2-containing EVs were able to bind the TLR2 ligand Pam3CSK4. In addition, the presence of EVs reduced inflammatory responses in Pam3CSK4-treated endothelial cells and HEK Dual reporter cells, demonstrating that TLR2-EVs can act as decoy receptors. In summary, our data show that sTLR2 and full-length TLR2 are released by macrophages under anti-inflammatory conditions, which may contribute to GC-induced immunosuppression.
Front Immunol. 2019 Jul 23;10:1634. doi: 10.3389/fimmu.2019.01634. eCollection 2019.
1664-3224
31396208
10.3389/fimmu.2019.01634
http://hdl.handle.net/10033/622122
Frontiers in Immunology
corticosteroid
exosome
innate immunity
microvesicle
pulmonary macrophage
Toll-Like Receptor 2 Release by Macrophages: An Anti-inflammatory Program Induced by Glucocorticoids and Lipopolysaccharide.
oai:repository.helmholtz-hzi.de:10033/6221292020-02-12T12:13:06Zcom_10033_620613com_10033_620618col_10033_620685col_10033_620619
00925njm 22002777a 4500
dc
Goes, Adriely
author
Lapuhs, Philipp
author
Kuhn, Thomas
author
Schulz, Eilien
author
Richter, Robert
author
Panter, Fabian
author
Dahlem, Charlotte
author
Koch, Marcus
author
Garcia, Ronald
author
Kiemer, Alexandra K
author
Müller, Rolf
author
Fuhrmann, Gregor
author
2020-01-12
In 2019, it was estimated that 2.5 million people die from lower tract respiratory infections annually. One of the main causes of these infections is Staphylococcus aureus, a bacterium that can invade and survive within mammalian cells. S. aureus intracellular infections are difficult to treat because several classes of antibiotics are unable to permeate through the cell wall and reach the pathogen. This condition increases the need for new therapeutic avenues, able to deliver antibiotics efficiently. In this work, we obtained outer membrane vesicles (OMVs) derived from the myxobacteria Cystobacter velatus strain Cbv34 and Cystobacter ferrugineus strain Cbfe23, that are naturally antimicrobial, to target intracellular infections, and investigated how they can affect the viability of epithelial and macrophage cell lines. We evaluated by cytometric bead array whether they induce the expression of proinflammatory cytokines in blood immune cells. Using confocal laser scanning microscopy and flow cytometry, we also investigated their interaction and uptake into mammalian cells. Finally, we studied the effect of OMVs on planktonic and intracellular S. aureus. We found that while Cbv34 OMVs were not cytotoxic to cells at any concentration tested, Cbfe23 OMVs affected the viability of macrophages, leading to a 50% decrease at a concentration of 125,000 OMVs/cell. We observed only little to moderate stimulation of release of TNF-alpha, IL-8, IL-6 and IL-1beta by both OMVs. Cbfe23 OMVs have better interaction with the cells than Cbv34 OMVs, being taken up faster by them, but both seem to remain mostly on the cell surface after 24 h of incubation. This, however, did not impair their bacteriostatic activity against intracellular S. aureus. In this study, we provide an important basis for implementing OMVs in the treatment of intracellular infections.
Cells. 2020 Jan 12;9(1). pii: cells9010194. doi: 10.3390/cells9010194.
2073-4409
31940898
10.3390/cells9010194
http://hdl.handle.net/10033/622129
Cells
Staphylococcus aureus
antimicrobial resistance
biogenic drug carriers
extracellular vesicles
intracellular infection
outer membrane vesicles
Myxobacteria-Derived Outer Membrane Vesicles: Potential Applicability Against Intracellular Infections.
oai:repository.helmholtz-hzi.de:10033/6221472020-02-19T02:01:47Zcom_10033_620613col_10033_620614col_10033_620685
00925njm 22002777a 4500
dc
Montefusco-Pereira, Carlos V.
author
Formicola, Beatrice
author
Goes, Adriely
author
Re, Francesca
author
Marrano, Claudia A.
author
Mantegazza, Francesco
author
Carvalho-Wodarz, Cristiane
author
Fuhrmann, Gregor
author
Caneva, Enrico
author
Nicotra, Francesco
author
Lehr, Claus-Michael
author
Russo, Laura
author
2020-04
By functionalizing the surface of PEG-liposomes with linkers bearing quaternary ammonium compounds (QACs), we generated novel bacteria disruptors with anti-adhesive properties and reduced cytotoxicity compared to free QACs. Furthermore, QAC-functionalized liposomes are a promising platform for future drug encapsulation. The QAC (11-mercaptoundecyl)-N,N,N-trimethylammonium bromide (MTAB) was attached to maleimide-functionalized liposomes (DSPE-PEG) via thiol linker. The MTAB-functionalized liposomes were physicochemically characterized and their biological activity, in terms of anti-adherence activity and biofilm prevention in Escherichia coli were assessed. The results showed that MTAB-functionalized liposomes inhibit bacterial adherence and biofilm formation while reducing MTAB toxicity.
Eur J Pharm Biopharm. 2020 Jan 30. pii: S0939-6411(20)30024-2. doi:10.1016/j.ejpb.2020.01.013.
0939-6411
32007589
10.1016/j.ejpb.2020.01.013
http://hdl.handle.net/10033/622147
European Journal of Pharmaceutics and Biopharmaceutics
Biotechnology
Pharmaceutical Science
General Medicine
Coupling quaternary ammonium surfactants to the surface of liposomes improves both antibacterial efficacy and host cell biocompatibility
oai:repository.helmholtz-hzi.de:10033/6222012020-03-20T03:34:34Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Fuhrmann, Gregor
author
2020-02-17
Cell-derived extracellular vesicles are important intercellular communicators involved in many biological processes and diseases, including cancer and cardiovascular diseases, but, thus far, how they navigate within complex extracellular matrices has been poorly understood.
Nat Nanotechnol. 2020 Feb 17. pii: 10.1038/s41565-020-0651-3. doi: 10.1038/s41565-020-0651-3.
1748-3395
32066903
10.1038/s41565-020-0651-3
http://hdl.handle.net/10033/622201
Nature nanotechnology
Diffusion and transport of extracellular vesicles.
oai:repository.helmholtz-hzi.de:10033/6222122020-03-19T02:02:59Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Mehanny, Mina
author
Koch, Marcus
author
Lehr, Claus-Michael
author
Fuhrmann, Gregor
author
2020-02-14
Extracellular vesicles are membranous structures shed by almost every living cell. Bacterial gram-negative outer membrane vesicles (OMVs) and gram-positive membrane vesicles (MVs) play important roles in adaptation to the surrounding environment, cellular components' exchange, transfer of antigens and virulence factors, and infection propagation. Streptococcus pneumoniae is considered one of the priority pathogens, with a global health impact due to the increase in infection burden and growing antibiotic resistance. We isolated MVs produced from the S. pneumoniae reference strain (R6) and purified them via size exclusion chromatography (SEC) to remove soluble protein impurities. We characterized the isolated MVs by nanoparticle tracking analysis (NTA) and measured their particle size distribution and concentration. Isolated MVs showed a mean particle size range of 130-160 nm and a particle yield of around 1012 particles per milliliter. Cryogenic transmission electron microscopy (cryo-TEM) images revealed a very heterogeneous nature of isolated MVs with a broad size range and various morphologies, arrangements, and contents. We incubated streptococcal MVs with several mammalian somatic cells, namely, human lung epithelial A549 and human keratinocytes HaCaT cell lines, and immune cells including differentiated macrophage-like dTHP-1 and murine dendritic DC2.4 cell lines. All cell lines displayed excellent viability profile and negligible cytotoxicity after 24-h incubation with MVs at concentrations reaching 106 MVs per cell (somatic cells) and 105 MVs per cell (immune cells). We evaluated the uptake of fluorescently labeled MVs into these four cell lines, using flow cytometry and confocal microscopy. Dendritic cells demonstrated prompt uptake after 30-min incubation, whereas other cell lines showed increasing uptake after 2-h incubation and almost complete colocalization/internalization of MVs after only 4-h incubation. We assessed the influence of streptococcal MVs on antigen-presenting cells, e.g., dendritic cells, using enzyme-linked immunosorbent assay (ELISA) and observed enhanced release of tumor necrosis factor (TNF)-α, a slight increase of interleukin (IL)-10 secretion, and no detectable effect on IL-12. Our study provides a better understanding of gram-positive streptococcal MVs and shows their potential to elicit a protective immune response. Therefore, they could offer an innovative avenue for safe and effective cell-free vaccination against pneumococcal infections.
Front Microbiol. 2020 Feb 7;11:124. doi: 10.3389/fmicb.2020.00124. eCollection 2020.
32117243
10.3389/fimmu.2020.00080
http://hdl.handle.net/10033/622212
1664-3224
Frontiers in immunology
Streptococcus pneumoniae
cytokine
cytotoxicity
extracellular membrane vesicles
uptake
vaccine
Streptococcal Extracellular Membrane Vesicles Are Rapidly Internalized by Immune Cells and Alter Their Cytokine Release.
oai:repository.helmholtz-hzi.de:10033/6222402020-04-24T01:34:59Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Zivko, Cristina
author
Fuhrmann, Gregor
author
Luciani, Paola
author
2020-02-19
BACKGROUND: Extracellular vesicles (EVs) are a diverse group of membrane-bound nanovesicles potentially released by every cell. With the liver's unique ensemble of cells and its fundamental physiological tasks, elucidating the role of EV-mediated hepatic cellular crosstalk and their role in different pathologies has been gaining the attention of many scientists.
SCOPE OF REVIEW: The present review shifts the perspective into practice: we aim to critically discuss the methods used to purify and to biochemically analyse EVs from specific liver resident cells, including hepatocytes, hepatic stellate cells, cholangiocytes, liver sinusoidal endothelial cells, Kupffer cells, liver stem cells. The review offers a reference guide to current approaches.
MAJOR CONCLUSIONS: Strategies for EV isolation and characterization are as varied as the research groups performing them. We present main advantages and disadvantages for the methods, highlighting common causes for concern, such as FBS handling, reporting of cell viability, EV yield and storage, differences in differential centrifugations, suboptimal method descriptions, and method transferability. We both looked at how adaptable the research between human and rodent cells in vitro is, and also assessed how well either of them translates to ex vivo settings.
GENERAL SIGNIFICANCE: We reviewed methodological practices for the isolation and analysis of liver-derived EVs, making a cell type specific user guide that shows where to start, what has worked so far and to what extent. We critically discussed room for improvement, placing a particular focus on working towards a potential standardization of methods.
Biochim Biophys Acta Gen Subj. 2020 Feb 19:129559. doi 10.1016/j.bbagen.2020.129559.
32084396
10.1016/j.bbagen.2020.129559
http://hdl.handle.net/10033/622240
1872-8006
Biochimica et biophysica acta. General subjects
Clinical translation
Exosomes
Liver cells
Microvesicles
PEGprecipitation
Ultracentrifugation
Liver-derived extracellular vesicles: A cell by cell overview to isolation and characterization practices.
oai:repository.helmholtz-hzi.de:10033/6223112020-06-25T01:28:29Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Goes, Adriely
author
Fuhrmann, Gregor
author
2018-03-29
Biogenic and biomimetic therapeutics are a relatively new class of systems that are of physiological origin and/or take advantage of natural pathways or aim at mimicking these to improve selective interaction with target tissue. The number of biogenic and bioengineered avenues for drug therapy and diagnostics has multiplied over the past years for many applications, indicating the high expectations associated with this biological route. Nevertheless, the use of "bio"-related approaches for treating or diagnosing infectious diseases is still rare. Given that infectious diseases, in particular bacterial resistances, are seriously on the rise, there is an urgent need to take advantage of biogenic and bioengineered systems to target these challenges. In this manuscript, we first give a definition of the various "bio" terms, including biogenic, biomimetic, bioinspired, and bioengineered and we highlight them using tangible applications in the field of infectious diseases. Our examples cover cell-derived systems, including bioengineered bacteria, virus-like particles, and different cell-mimetics. Moreover, we discuss natural and bioengineered particles such as extracellular vesicles from mammalian and bacterial sources and liposomes. A concluding section outlines the potential for biomaterial-related avenues to overcome challenges associated with difficult-to-treat infections. We critically discuss benefits and risks for these applications and give an outlook on the future of biogenic engineering.
ACS Infect Dis. 2018;4(6):881-892. doi:10.1021/acsinfecdis.8b00030.
29553240
10.1021/acsinfecdis.8b00030
http://hdl.handle.net/10033/622311
2373-8227
ACS infectious diseases
biogenic drug delivery
bioinspired delivery systems
biomimetics
cell-mimetics
extracellular vesicles
nanoantibiotics
outer membrane vesicles
virus-like particles
Biogenic and Biomimetic Carriers as Versatile Transporters To Treat Infections.
oai:repository.helmholtz-hzi.de:10033/6224492020-09-25T03:25:33Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Kuhn, Thomas
author
Koch, Marcus
author
Fuhrmann, Gregor
author
2020-09-03
There is a lack of efficient therapies to treat increasingly prevalent autoimmune diseases, such as inflammatory bowel disease and celiac disease. Membrane vesicles (MVs) isolated from probiotic bacteria have shown tremendous potential for treating intestinal inflammatory diseases. However, possible dilution effects and rapid elimination in the gastrointestinal tract may impair their application. A cell‐free and anti‐inflammatory therapeutic system—probiomimetics—based on MVs of probiotic bacteria (Lactobacillus casei and Lactobacillus plantarum) coupled to the surface of microparticles is developed. The MVs are isolated and characterized for size and protein content. MV morphology is determined using cryoelectron microscopy and is reported for the first time in this study. MVs are nontoxic against macrophage‐like dTHP‐1 and enterocyte‐like Caco‐2 cell lines. Subsequently, the MVs are coupled onto the surface of microparticles according to facile aldehyde‐group functionalization to obtain probiomimetics. A significant reduction in proinflammatory TNF‐α level (by 86%) is observed with probiomimetics but not with native MVs. Moreover, it is demonstrated that probiomimetics have the ability to ameliorate inflammation‐induced loss of intestinal barrier function, indicating their potential for further development into an anti‐inflammatory formulation. These engineered simple probiomimetics that elicit striking anti‐inflammatory effects are a key step toward therapeutic MV translation.
Small. 2020 Sep 3:e2003158. doi: 10.1002/smll.202003158. Epub ahead of print. PMID: 32885611.
32885611
10.1002/smll.202003158
http://hdl.handle.net/10033/622449
1613-6829
Small (Weinheim an der Bergstrasse, Germany)
anti-inflammatory therapy
bacteriomimetics
biomimetics
inflammatory bowel diseases
Probiomimetics-Novel Lactobacillus-Mimicking Microparticles Show Anti-Inflammatory and Barrier-Protecting Effects in Gastrointestinal Models.
oai:repository.helmholtz-hzi.de:10033/6225132020-10-16T01:44:58Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Whitworth, David E
author
Jurkevitch, Edouard
author
Pérez, Juana
author
Fuhrmann, Gregor
author
Koval, Susan F
author
2020-09-08
No abstract available]
Front Microbiol. 2020 Sep 8;11:2071. doi: 10.3389/fmicb.2020.02071.
1664-302X
33013755
10.3389/fmicb.2020.02071
http://hdl.handle.net/10033/622513
Frontiers in microbiology
BALO
Bdellovibrio
microbial ecology
myxobacteria
prey
Editorial: Mechanisms of Prokaryotic Predation.
oai:repository.helmholtz-hzi.de:10033/6226132020-12-04T03:37:40Zcom_10033_620613col_10033_620614col_10033_620685
00925njm 22002777a 4500
dc
Fuhrmann, Gregor
author
Loretz, Brigitta
author
Schneider-Daum, Nicole
author
Lehr, Claus-Michael
author
2020-11-02
[No abstract available]
Eur J Pharm Biopharm. 2020 Nov 2:S0939-6411(20)30314-3. doi: 10.1016/j.ejpb.2020.10.014. Epub ahead of print.
33152481
10.1016/j.ejpb.2020.10.014
http://hdl.handle.net/10033/622613
1873-3441
European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V
Editorial EJPB - Biobarriers 2018.
oai:repository.helmholtz-hzi.de:10033/6228402021-04-24T01:45:13Zcom_10033_620613col_10033_620614col_10033_620685
00925njm 22002777a 4500
dc
Mehanny, Mina
author
Lehr, Claus-Michael
author
Fuhrmann, Gregor
author
2021-03-26
Antigen delivery has always been a challenge in scientific practice of vaccine formulation. Yet, mammalian extracellular vesicles (EVs) or bacterial membrane vesicles (MVs) provide an innovative avenue for safe and effective delivery of antigenic material. They include intrinsically loaded antigens from EV-secreting cells or extrinsically loaded antigens onto pre-formed vesicles. Interestingly, many studies shed light on potential novel anti-cancer vaccination immunotherapy for therapeutic applications from mammalian cell host-derived EVs, as well as conventional vaccination for prophylactic applications using bacterial cell-derived MVs against infectious diseases. Here, we discuss the rationale, status quo and potential for both vaccine applications using EVs.
Adv Drug Deliv Rev. 2021 Mar 26;173:164-180. doi: 10.1016/j.addr.2021.03.016. Epub ahead of print.
33775707
10.1016/j.addr.2021.03.016
http://hdl.handle.net/10033/622840
1872-8294
Advanced drug delivery reviews
Antigen
Cancer
Extracellular vesicles
Immunotherapy
Infection
Outer membrane vesicles
Vaccine
Extracellular vesicles as antigen carriers for novel vaccination avenues.
oai:repository.helmholtz-hzi.de:10033/6228412021-04-24T01:45:13Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Jahromi, Leila Pourtalebi
author
Fuhrmann, Gregor
author
2021-03-25
Extracellular vesicle (EV)-mediated communication between proximal and distant cells is a highly conserved characteristic in all of the life domains, including bacteria. These vesicles that contain a variety of biomolecules, such as proteins, lipids, nucleic acids, and small-molecule metabolites play a key role in the biology of bacteria. They are one of the key underlying mechanisms behind harmful or beneficial effects of many pathogenic, symbiont, and probiotic bacteria. These nanoscale EVs mediate extensive crosstalk with mammalian cells and deliver their cargos to the host. They are stable in physiological condition, can encapsulate diverse biomolecules and nanoparticles, and their surface could be engineered with available technologies. Based on favorable characteristics of bacterial vesicles, they can be harnessed for designing a diverse range of therapeutics and diagnostics for treatment of disorders including tumors and resistant infections. However, technical limitations for their production, purification, and characterization must be addressed in future studies.
Adv Drug Deliv Rev. 2021 Mar 25;173:125-140. doi: 10.1016/j.addr.2021.03.012. Epub ahead of print.
33774113
10.1016/j.addr.2021.03.012
http://hdl.handle.net/10033/622841
1872-8294
Advanced drug delivery reviews
Antimicrobials
Bacterial extracellular vesicles
Bioengineering
Biogenic nanotherapeutics
Bioinspired
Drug delivery
Outer membrane vesicles
Bacterial extracellular vesicles: Understanding biology promotes applications as nanopharmaceuticals.
oai:repository.helmholtz-hzi.de:10033/6228602021-05-08T01:43:51Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Richter, Maximilian
author
Vader, Pieter
author
Fuhrmann, Gregor
author
2021-04-06
Extracellular vesicles (EVs) are cell-derived nanoparticles that are important mediators in intercellular communication. This function makes them auspicious candidates for therapeutic and drug-delivery applications. Among EVs, mammalian cell derived EVs and outer membrane vesicles (OMVs) produced by gram-negative bacteria are the most investigated candidates for pharmaceutical applications. To further optimize their performance and to utilize their natural abilities, researchers have strived to equip EVs with new moieties on their surface while preserving the integrity of the vesicles. The aim of this review is to give a comprehensive overview of techniques that can be used to introduce these moieties to the vesicle surface. Approaches can be classified in regards to whether they take place before or after the isolation of EVs. The producing cells can be subjected to genetic manipulation or metabolic engineering to produce surface modified vesicles or EVs are engineered after their isolation by physical or chemical means. Here, the advantages and disadvantages of these processes and their applicability for the development of EVs as therapeutic agents are discussed.
Adv Drug Deliv Rev. 2021 Apr 6;173:416-426. doi: 10.1016/j.addr.2021.03.020. Epub ahead of print.
33831479
10.1016/j.addr.2021.03.020
http://hdl.handle.net/10033/622860
1872-8294
Advanced drug delivery reviews
Drug delivery
Exosomes
Extracellular vesicles
Fluorescent labeling
Outer membrane vesicles
Surface functionalization
Targeting
Approaches to surface engineering of extracellular vesicles.
oai:repository.helmholtz-hzi.de:10033/6229662021-07-29T01:53:34Zcom_10033_620613col_10033_620685
00925njm 22002777a 4500
dc
Herrmann, Inge Katrin
author
Wood, Matthew John Andrew
author
Fuhrmann, Gregor
author
2021-07-01
Extracellular-vesicle-based cell-to-cell communication is conserved across all kingdoms of life. There is compelling evidence that extracellular vesicles are involved in major (patho)physiological processes, including cellular homoeostasis, infection propagation, cancer development and cardiovascular diseases. Various studies suggest that extracellular vesicles have several advantages over conventional synthetic carriers, opening new frontiers for modern drug delivery. Despite extensive research, clinical translation of extracellular-vesicle-based therapies remains challenging. Here, we discuss the uniqueness of extracellular vesicles along with critical design and development steps required to utilize their full potential as drug carriers, including loading methods, in-depth characterization and large-scale manufacturing. We compare the prospects of extracellular vesicles with those of the well established liposomes and provide guidelines to direct the process of developing vesicle-based drug delivery systems.
(2021).Nature Nanotechnology, 16(7), 748-759. doi:10.1038/s41565-021-00931-2.
34211166
10.1038/s41565-021-00931-2
http://hdl.handle.net/10033/622966
1748-3395
Nature nanotechnology
Extracellular vesicles as a next-generation drug delivery platform.
oai:repository.helmholtz-hzi.de:10033/6231392022-01-13T01:55:10Zcom_10033_620613com_10033_620589com_10033_338554com_10033_620636col_10033_621787col_10033_620685col_10033_620590col_10033_620638
00925njm 22002777a 4500
dc
Goes, Adriely
author
Vidakovic, Lucia
author
Drescher, Knut
author
Fuhrmann, Gregor
author
2021-08-02
Bacterial biofilms are widespread in nature and in medical settings and display a high tolerance to antibiotics and disinfectants. Extracellular vesicles have been increasingly studied to characterise their origins and assess their potential for use as a versatile drug delivery system; however, it remains unclear whether they also have antibiofilm effects. Outer membrane vesicles are lipid vesicles shed by Gram-negative bacteria and, in the case of myxobacteria, carry natural antimicrobial compounds produced by these microorganisms. In this study, we demonstrate that vesicles derived from the myxobacteria Cystobacter velatus Cbv34 and Cystobacter ferrugineus Cbfe23 are highly effective at inhibiting the formation and disrupting biofilms by different bacterial species.
34477714
10.1039/d1nr02583j
http://hdl.handle.net/10033/623139
2040-3372
Nanoscale
Interaction of myxobacteria-derived outer membrane vesicles with biofilms: antiadhesive and antibacterial effects.
oai:repository.helmholtz-hzi.de:10033/6231632022-02-12T01:55:14Zcom_10033_620613com_10033_620656col_10033_620685col_10033_620657
00925njm 22002777a 4500
dc
Drost, Menka
author
Diamanti, Eleonora
author
Fuhrmann, Kathrin
author
Goes, Adriely
author
Shams, Atanaz
author
Haupenthal, Jörg
author
Koch, Marcus
author
Hirsch, Anna K H
author
Fuhrmann, Gregor
author
2021-12-21
Liposomes have been studied for decades as nanoparticulate drug delivery systems for cytostatics, and more recently, for antibiotics. Such nanoantibiotics show improved antibacterial efficacy compared to the free drug and can be effective despite bacterial recalcitrance. In this work, we present a loading method of bacteriomimetic liposomes for a novel, hydrophobic compound (HIPS5031) inhibiting energy-coupling factor transporters (ECF transporters), an underexplored antimicrobial target. The liposomes were composed of DOPG (18:1 (Δ9-cis) phosphatidylglycerol) and CL (cardiolipin), resembling the cell membrane of Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and enriched with cholesterol (Chol). The size and polydispersity of the DOPG/CL/± Chol liposomes remained stable over 8 weeks when stored at 4 °C. Loading of the ECF transporter inhibitor was achieved by thin film hydration and led to a high encapsulation efficiency of 33.19% ± 9.5% into the DOPG/CL/Chol liposomes compared to the phosphatidylcholine liposomes (DMPC/DPPC). Bacterial growth inhibition assays on the model organism Bacillus subtilis revealed liposomal HIPS5031 as superior to the free drug, showing a 3.5-fold reduction in CFU/mL at a concentration of 9.64 µM. Liposomal HIPS5031 was also shown to reduce B. subtilis biofilm. Our findings present an explorative basis for bacteriomimetic liposomes as a strategy against drug-resistant pathogens by surpassing the drug-formulation barriers of innovative, yet unfavorably hydrophobic, antibiotics.
Pharmaceutics. 2021 Dec 21;14(1):4. doi: 10.3390/pharmaceutics14010004.
1999-4923
35056900
10.3390/pharmaceutics14010004
http://hdl.handle.net/10033/623163
Pharmaceutics
Bacillus subtilis
antibiotic resistance
bacteriomimetic
cardiolipin
energy-coupling factor (ECF) transporters
liposomes
nanoantibiotics
Bacteriomimetic Liposomes Improve Antibiotic Activity of a Novel Energy-Coupling Factor Transporter Inhibitor.