| 1. |
- Boge, Lukas, et al.
(författare)
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Peptide-Loaded Cubosomes Functioning as an Antimicrobial Unit against Escherichia coli
- 2019
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Ingår i: ACS Applied Materials and Interfaces. - : American Chemical Society. - 1944-8244 .- 1944-8252. ; 11:24, s. 21314-21322
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Tidskriftsartikel (refereegranskat)abstract
- Dispersions of cubic liquid crystalline phases, also known as cubosomes, have shown great promise as delivery vehicles for a wide range of medicines. Due to their ordered structure, comprising alternating hydrophilic and hydrophobic domains, cubosomes possess unique delivery properties and compatibility with both water-soluble and -insoluble drugs. However, the drug delivery mechanism and cubosome interaction with human cells and bacteria are still poorly understood. Herein, we reveal how cubosomes loaded with the human cathelicidin antimicrobial peptide LL-37, a system with high bacteria-killing effect, interact with the bacterial membrane and provide new insights into the eradication mechanism. Combining the advanced experimental techniques neutron reflectivity and quartz crystal microbalance with dissipation monitoring, a mechanistic drug delivery model for LL-37-loaded cubosomes on bacterial mimicking bilayers was constructed. Moreover, the cubosome interaction with Escherichia coli was directly visualized using super-resolution laser scanning microscopy and cryogenic electron tomography. We could conclude that cubosomes loaded with LL-37 adsorbed and distorted bacterial membranes, providing evidence that the peptide-loaded cubosomes function as an antimicrobial unit.
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| 2. |
- Borro, Bruno C., et al.
(författare)
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Microgels and hydrogels as delivery systems for antimicrobial peptides
- 2020
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Ingår i: Colloids and Surfaces B. - : ELSEVIER. - 0927-7765 .- 1873-4367. ; 187
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Tidskriftsartikel (refereegranskat)abstract
- Due to rapid development of bacterial resistance against antibiotics, an emerging health crisis is underway, where `simple' infections may no longer be treatable. Antimicrobial peptides (AMPs) constitute a class of substances attracting interest in this context. So far, research on AMPs has primarily focused on the identification of potent and selective peptides, as well as on the action mode of such peptides. More recently, there has been an increasing awareness that the delivery of AMPs is challenging due to their size, net positive charge, amphiphilicity, and proteolytic susceptibility. Hence, successful development of AMP therapeutics will likely require also careful design of efficient AMP delivery systems. In the present brief review, we discuss microgels, as well as related polyelectrolyte complexes and macroscopic hydrogels, as delivery systems for AMPs. In doing so, key factors for peptide loading and release are outlined and exemplified, together with consequences of this for functional performance relating to antimicrobial effects and cell toxicity.
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| 3. |
- Braun, Katharina, et al.
(författare)
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Membrane interactions of mesoporous silica nanoparticles as carriers of antimicrobial peptides
- 2016
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 475, s. 161-170
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Tidskriftsartikel (refereegranskat)abstract
- Membrane interactions are critical for the successful use of mesoporous silica nanoparticles as delivery systems for antimicrobial peptides (AMPs). In order to elucidate these, we here investigate effects of nanoparticle charge and porosity on AMP loading and release, as well as consequences of this for membrane interactions and antimicrobial effects. Anionic mesoporous silica particles were found to incorporate considerable amounts of the cationic AMP LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES (LL-37), whereas loading is much lower for non-porous or positively charged silica nanoparticles. Due to preferential pore localization, anionic mesoporous particles, but not the other particles, protect LL-37 from degradation by infection-related proteases. For anionic mesoporous nanoparticles, membrane disruption is mediated almost exclusively by peptide release. In contrast, non-porous silica particles build up a resilient LL-37 surface coating due to their higher negative surface charge, and display largely particle-mediated membrane interactions and antimicrobial effects. For positively charged mesoporous silica nanoparticles, LL-37 incorporation promotes the membrane binding and disruption displayed by the particles in the absence of peptide, but also causes toxicity against human erythrocytes. Thus, the use of mesoporous silica nanoparticles as AMP delivery systems requires consideration of membrane interactions and selectivity of both free peptide and the peptide-loaded nanoparticles, the latter critically dependent on nanoparticle properties.
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| 4. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Membrane interactions and antimicrobial effects of layered double hydroxide nanoparticles
- 2017
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Ingår i: Physical Chemistry, Chemical Physics - PCCP. - : ROYAL SOC CHEMISTRY. - 1463-9076 .- 1463-9084. ; 19:35, s. 23832-23842
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Tidskriftsartikel (refereegranskat)abstract
- Membrane interactions are critical for the successful use of inorganic nanoparticles as antimicrobial agents and as carriers of, or co-actives with, antimicrobial peptides (AMPs). In order to contribute to an increased understanding of these, we here investigate effects of particle size (42-208 nm) on layered double hydroxide (LDH) interactions with both bacteria-mimicking and mammalian-mimicking lipid membranes. LDH binding to bacteria-mimicking membranes, extraction of anionic lipids, as well as resulting membrane destabilization, was found to increase with decreasing particle size, also translating into size-dependent synergistic effects with the antimicrobial peptide LL-37. Due to strong interactions with anionic lipopolysaccharide and peptidoglycan layers, direct membrane disruption of both Gram-negative and Gram-positive bacteria is suppressed. However, LDH nanoparticles cause size-dependent charge reversal and resulting flocculation of both liposomes and bacteria, which may provide a mechanism for bacterial confinement or clearance. Taken together, these findings demonstrate a set of previously unknown behaviors, including synergistic membrane destabilization and dual confinement/killing of bacteria through combined LDH/AMP exposure, of potential therapeutic interest.
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| 5. |
- Nordström, Randi, 1986-, et al.
(författare)
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Degradable dendritic nanogels as carriers for antimicrobial peptides
- 2019
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 554, s. 592-602
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, we investigate degradable anionic dendritic nanogels (DNG) as carriers for antimicrobial peptides (AMPs). In such systems, the dendritic part contains carboxylic acid-based anionic binding sites for cationic AMPs, whereas linear poly(ethylene glycol) (PEG) chains form a shell for promotion of biological stealth. In order to clarify factors influencing membrane interactions of such systems, we here address effects of nanogel charge, cross-linking, and degradation on peptide loading/release, as well as consequences of these factors for lipid membrane interactions and antimicrobial effects. The DNGs were found to bind the AMPs LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) and DPK-060 (GKHKNKGKKNGKHNGWKWWW). For the smaller DPK-060 peptide, loading was found to increase with increasing nanogel charge density. For the larger LL-37, on the other hand, peptide loading was largely insensitive to nanogel charge density. In line with this, results on the secondary structure, as well as on the absence of stabilization from proteolytic degradation by the nanogels, show that the larger LL-37 is unable to enter into the interior of the nanogels. While 40–60% nanogel degradation occurred over 10 days, promoted at high ionic strength and lower cross-linking density/higher anionic charge content, peptide release at physiological ionic strength was substantially faster, and membrane destabilization not relying on nanogel degradation. Ellipsometry and liposome leakage experiments showed both free peptide and peptide/DNG complexes to cause membrane destabilization, indicated also by antimicrobial activities being comparable for nanogel-bound and free peptide. Finally, the DNGs were demonstrated to display low toxicity towards erythrocytes even at peptide concentrations of 100 µM.
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| 6. |
- Nordström, Randi, et al.
(författare)
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Delivery systems for antimicrobial peptides
- 2017
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Ingår i: Advances in Colloid and Interface Science. - : ELSEVIER SCIENCE BV. - 0001-8686 .- 1873-3727. ; 242, s. 17-34
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Tidskriftsartikel (refereegranskat)abstract
- Due to rapidly increasing resistance development against conventional antibiotics, finding novel approaches for the treatment of infections has emerged as a key health issue. Antimicrobial peptides (AMPs) have attracted interest in this context, and there is by now a considerable literature on the identification such peptides, as well as on their optimization to reach potent antimicrobial and anti-inflammatory effects at simultaneously low toxicity against human cells. In comparison, delivery systems for antimicrobial peptides have attracted considerably less interest. However, such delivery systems are likely to play a key role in the development of potent and safe AMP based therapeutics, e.g., through reducing chemical or biological degradation of AMPs either in the formulation or after administration, by reducing adverse side-effects, by controlling AMP release rate, by promoting biofilm penetration, or through achieving co-localization with intracellular pathogens. Here, an overview is provided of the current understanding of delivery systems for antimicrobial peptides, with special focus on AMP-carrier interactions, as well as consequences of these interactions for antimicrobial and related biological effects of AMP-containing formulations.
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| 7. |
- Nordström, Randi, et al.
(författare)
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Membrane Interactions of Antimicrobial Peptide-Loaded Microgels
- 2020
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 562, s. 322-332
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, lipid membrane interactions of anionic poly(ethyl acrylate-co-methacrylic acid) (MAA) microgels as carriers for the cationic antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) were investigated. In doing so, neutron reflectometry (NR), Fourier-transform infrared spectroscopy with attenuated total reflection (FTIR-ATR), zeta potential, ellipsometry, and circular dichroism spectroscopy (CD) experiments were employed to investigate the relative importance of membrane interactions of peptide-loaded microgel particles and of released peptide. For the free peptide, NR results showed membrane binding occurring preferentially in the tail region in a concentration-dependent manner. At low peptide concentration (0.3 mu M) only peptide insertion in the outer leaflet was seen, however, pronounced membrane defects and peptide present in both leaflets was observed at higher peptide concentration (5.0 LL-37 loaded into MAA microgels qualitatively mirrored these effects regarding both peptide localization within the membrane and concentration dependent defect formation. In addition, very limited membrane binding of microgel particles was observed, in agreement with FTIR-ATR and liposome leakage results. FTIR-ATR showed LL-37 to undergo alpha-helix formation on membrane insertion, also supported by CD results, the kinetics of which was substantially reduced for microgel-loaded LL-37 due to sustained peptide release. Together, these findings demonstrate that membrane interactions for microgel-loaded LL-37 are dominated by released peptide, but also that slow release of microgel-loaded LL-37 translates into kinetic effects on peptide-membrane interactions, relating to both peptide localization within the bilayer, and to bilayer structure.
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| 8. |
- Nordström, Randi, et al.
(författare)
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Membrane interactions of microgels as carriers of antimicrobial peptides
- 2018
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Ingår i: Journal of Colloid and Interface Science. - : Academic Press Inc.. - 0021-9797 .- 1095-7103. ; 513, s. 141-150
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Tidskriftsartikel (refereegranskat)abstract
- Microgels are interesting as potential delivery systems for antimicrobial peptides. In order to elucidate membrane interactions of such systems, we here investigate effects of microgel charge density on antimicrobial peptide loading and release, as well as consequences of this for membrane interactions and antimicrobial effects, using ellipsometry, circular dichroism spectroscopy, nanoparticle tracking analysis, dynamic light scattering and z-potential measurements. Anionic poly(ethyl acrylate-co-methacrylic acid) microgels were found to incorporate considerable amounts of the cationic antimicrobial peptides LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) and DPK-060 (GKHKNKGKKNGKHNGWKWWW) and to protect incorporated peptides from degradation by infection-related proteases at high microgel charge density. As a result of their net negative z-potential also at high peptide loading, neither empty nor peptide-loaded microgels adsorb at supported bacteria-mimicking membranes. Instead, membrane disruption is mediated almost exclusively by peptide release. Mirroring this, antimicrobial effects against several clinically relevant bacteria (methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, and Pseudomonas aeruginosa) were found to be promoted by factors facilitating peptide release, such as decreasing peptide length and decreasing microgel charge density. Microgels were further demonstrated to display low toxicity towards erythrocytes. Taken together, the results demonstrate some interesting opportunities for the use of microgels as delivery systems for antimicrobial peptides, but also highlight several key factors which need to be controlled for their successful use.
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| 9. |
- Nordström, Randi, et al.
(författare)
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Microgels as carriers of antimicrobial peptides – effects of peptide PEGylation
- 2019
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Ingår i: Colloids and Surfaces A. - : Elsevier BV. - 0927-7757 .- 1873-4359. ; 565, s. 8-15
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Tidskriftsartikel (refereegranskat)abstract
- Delivery systems are likely to be central for the translation of antimicrobial peptides (AMPs) towards therapeutics. Addressing AMP interactions with microgel carriers, we here investigate how poly(ethylene glycol) conjugation ('PEGylation') of AMPs affect their loading and release to/from microgels, combining structural studies using nuclear magnetic resonance (NMR) with ellipsometry, circular dichroism spectroscopy (CD), and light scattering. Such studies demonstrate that poly(ethyl acrylate-co-methacrylic acid) microgels bind considerable amounts of the positively charged AMP KYE28 (KYEITTIHNLFRKLTHRLFRRNFGYTLR) and its PEGylated variants KYE28-PEG48, PEG48-KYE28, and PEG24-KYE28-PEG24. Z-potential measurements indicate that KYE28 resides primarily inside the microgel core, and that localization of the PEGylated peptides is shifted towards the microgel corona. Furthermore, while all peptides are disordered in solution, CD measurements report on helix induction on microgel binding, particularly so for the PEGylated peptides. Addressing such conformational changes in more detail, NMR structural studies showed that peptide-microgel interactions are facilitated by a hydrophobic domain formed by the peptide after microgel binding, and with modulating electrostatic/salt bridge interaction between the positively charged peptide residues and negative microgel charges. As the microgels remain negatively charged also at high peptide load, membrane disruption and antimicrobial effects necessitates peptide release, demonstrated to be promoted by PEGylation and high ionic strength. Importantly, microgel loading, as well as peptide localization, conformation, and release, did not depend significantly on PEG conjugation site, but instead seems to be dictated by the PEG content of the peptide conjugates.
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| 10. |
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| 11. |
- Nordström, Randi
(författare)
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Polymeric Nanoparticles as Carriers for Antimicrobial Peptides : Factors Affecting Peptide and Membrane Interactions
- 2019
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- As resistance towards conventional antibiotics is becoming more pronounced, cationic antimicrobial peptides (AMPs) have received considerable attention as possible therapeutic alternatives. Thousands of potent AMPs occur in humans, animals, plants and fungi as a natural part of the immune system. However, there are several challenges with AMP therapeutics related to formulation and delivery. Examples include proteolytic sensitivity and serum protein binding, resulting in quick degradation, loss of activity and clearance. Therefore, it is important to find a suitable drug delivery system to meet these protection and delivery challenges. Micro-/nanogels are loosely crosslinked polymer colloids with high water content that can be made to trigger at a wide range of stimuli. They have shown promise as delivery systems for AMPs, as the aqueous environment they create allows the peptides to maintain their natural conformation, while their gel networks offer protection and triggered release. This thesis aims towards expanding the knowledge about degradable and non-degradable pH-responsive micro-/nanogels as carriers for AMPs.The results in this thesis show that factors relating to the drug delivery system (degradability, charge and crosslinker density), the surrounding media (pH and ionic strength) and the peptide properties (length, charge, PEGylation) all affect the peptide loading to, protection, release from and effect of AMP-loaded gels. Studies of the interaction of AMP-loaded microgels with bacteria-modelling liposomes and lipid bilayers have verified peptide effect after gel incorporation, as further demonstrated by in vitro studies on several bacterial strains. Neutron reflectometry provided detailed mechanistic information on the interaction between AMP-loaded gels and bacteria-modelling lipid bilayers, showing that the antimicrobial unit is the released peptide. All gels showed low, promising hemolysis and some gels could offer protection against proteolytic degradation of AMPs.In summary, non-degradable and degradable micro-/nanogels are versatile and interesting candidates as AMP carriers. Small changes in the gel composition or the AMP used can dramatically change the peptide loading, release and effect. It is therefore necessary to carefully consider and evaluate the optimal carrier for every AMP and the application at hand.
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| 12. |
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| 13. |
- Nyström, Lina, et al.
(författare)
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Factors affecting peptide interactions with surface-bound microgels
- 2016
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Ingår i: Biomacromolecules. - : American Chemical Society (ACS). - 1525-7797 .- 1526-4602. ; 17:2, s. 669-678
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Tidskriftsartikel (refereegranskat)abstract
- Effects of electrostatics and peptide size on peptide interactions with surface-bound microgels were investigated with ellipsometry, confocal microscopy, and atomic force microscopy (AFM). Results show that binding of cationic poly-l-lysine (pLys) to anionic, covalently immobilized, poly(ethyl acrylate-co-methacrylic acid) microgels increased with increasing peptide net charge and microgel charge density. Furthermore, peptide release was facilitated by decreasing either microgel or peptide charge density. Analogously, increasing ionic strength facilitated peptide release for short peptides. As a result of peptide binding, the surface-bound microgels displayed pronounced deswelling and increased mechanical rigidity, the latter quantified by quantitative nanomechanical mapping. While short pLys was found to penetrate the entire microgel network and to result in almost complete charge neutralization, larger peptides were partially excluded from the microgel network, forming an outer peptide layer on the microgels. As a result of this difference, microgel flattening was more influenced by the lower Mw peptide than the higher. Peptide-induced deswelling was found to be lower for higher Mw pLys, the latter effect not observed for the corresponding microgels in the dispersed state. While the effects of electrostatics on peptide loading and release were similar to those observed for dispersed microgels, there were thus considerable effects of the underlying surface on peptide-induced microgel deswelling, which need to be considered in the design of surface-bound microgels as carriers of peptide loads, for example, in drug delivery or in functionalized biomaterials.
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| 14. |
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| 15. |
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| 16. |
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| 17. |
- Parra-Ortiz, Elisa, et al.
(författare)
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Effects of oxidation on the physicochemical properties of polyunsaturated lipid membranes
- 2019
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Ingår i: Journal of Colloid and Interface Science. - : ACADEMIC PRESS INC ELSEVIER SCIENCE. - 0021-9797 .- 1095-7103. ; 538, s. 404-419
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Tidskriftsartikel (refereegranskat)abstract
- The exposure of biological membranes to reactive oxygen species (ROS) plays an important role in many pathological conditions such as inflammation, infection, or sepsis. ROS also modulate signaling processes and produce markers for damaged tissue. Lipid peroxidation, mainly affecting polyunsaturated phospholipids, results in a complex mixture of oxidized products, which may dramatically alter membrane properties. Here, we have employed a set of biophysical and surface-chemical techniques, including neutron and X-ray scattering, to study the structural, compositional, and stability changes due to oxidative stress on phospholipid bilayers composed of lipids with different degrees of polyunsaturation. In doing so, we obtained real-time information about bilayer degradation under in situ UV exposure using neutron reflectometry. We present a set of interrelated physicochemical effects, including gradual increases in area per molecule, head group and acyl chain hydration, as well as bilayer thinning, lateral phase separation, and defect formation leading to content loss upon membrane oxidation. Such effects were observed to depend on the presence of polyunsaturated phospholipids in the lipid membrane, suggesting that these may also play a role in the complex oxidation processes occurring in cells.
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| 18. |
- Zhang, Yuning, et al.
(författare)
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Off-Stoichiometric Thiol-Ene Chemistry to Dendritic Nanogel Therapeutics
- 2019
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Ingår i: Advanced Functional Materials. - : Wiley-VCH Verlag. - 1616-301X .- 1616-3028. ; 29:18
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Tidskriftsartikel (refereegranskat)abstract
- A novel platform of dendritic nanogels is herein presented, capitalizing on the self-assembly of allyl-functional polyesters based on dendritic-linear-dendritic amphiphiles followed by simple cross-linking with complementary monomeric thiols via UV initiated off-stoichiometric thiol-ene chemistry. The facile approach enabled multigram creation of allyl reactive nanogel precursors, in the size range of 190–295 nm, being readily available for further modifications to display a number of core functionalities while maintaining the size distribution and characteristics of the master batch. The nanogels are evaluated as carriers of a spread of chemotherapeutics by customizing the core to accommodate each individual cargo. The resulting nanogels are biocompatible, displaying diffusion controlled release of cargo, maintained therapeutic efficacy, and decreased cargo toxic side effects. Finally, the nanogels are found to successfully deliver pharmaceuticals into a 3D pancreatic spheroids tumor model.
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