1. |
- Jing, Yujia, 1985
(författare)
-
Hyperthermia-responsive liposomal systems
- 2015
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Abstract Sophisticated liposomal systems are emerging at an increasing rate to meet the demands for multifunctional drug carriers in chemotherapies in combined with hyperthermia. For example, liposomal drug carriers for temperature-controlled drug release under hyperthermic conditions have recently been tested in clinical trials. More advanced designs of liposomes are expected to release encapsulated contents and activate hidden surface-functions in response to heat stimulus. Towards this aim, the present thesis is focused on formulating asymmetric lipid systems that can preserve functional moieties, and reactivate the targeted function as well as release the encapsulated compounds upon local heating. The design of the asymmetric liposomal systems utilizes the heat-activated transmembrane lipid diffusion during gel to liquid-crystalline phase transitions of the lipid membranes.Rational design of advanced liposomal drug-delivery systems will require understanding of the physicochemical properties of lipid membranes under, e.g., hyperthermic conditions. Here, supported lipid membranes on planar solid surfaces were used for model studies of lipid composition yielding a gel to liquid crystalline phase-transition temperature in the range 40 – 45 °C. It was found that the liposome-to-membrane formation process is not only size-dependent but also governed by temperature. Two methods of preparing supported asymmetric lipid membranes were investigated. As a proof-of-concept, the upper leaflets were either replaced or chemically transformed by enzymatic hydrolysis. The processes were monitored using surface sensitive techniques such as quartz crystal microbalance with dissipation (QCM-D) and dual polarization interferometry (DPI). The asymmetric structures were stable at a room temperature, while lipid flip-flop was induced upon increasing of the temperature. Transmembrane lipid exchange in the asymmetric structure under hyperthermic conditions was demonstrated by detecting, through streptavidin binding, biotinylated lipids appearing at the top leaflet which were first located in the lower leaflet. The protocols developed for the supported lipid systems were adapted for the preparation of asymmetric liposomes. Biotinylated asymmetric liposomes were used as a model system to demonstrate the principle of heat-activated targeting of asymmetric liposomes to streptavidin-coated surfaces. More biologically relevant interaction was utilized to replace the biotin-streptavidin function, where asymmetric cationic liposomes were binding to anionic supported membrane immobilized surfaces upon heating. The described strategies for assembly of asymmetric supported membranes provide a guide to the development of multifunctional drug carriers. The protocols used in experiments with supported membranes were readily adapted to the preparation of asymmetric liposomes. The ongoing study tests the asymmetric liposomes in vitro, which is designed to demonstrate hyperthermia treatment can enhance accumulation of liposomes in FaDu cells, and at the same time activate release of the encapsulated components. The results of in vitro tests can be used to analyze the feasibility of utilizing the asymmetric liposomes as a platform in vivo to explore further improvement in their functions upon microwave hyperthermia.
|
|
2. |
- Orru, Anna Maria, 1976, et al.
(författare)
-
AHA! festival 2015
- 2015
-
Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- The AHA festival investigates the borders between art and science in a three-day event at the Chalmers University of Technology hosted by the Department of Architecture. An international festival intended to provide enlightening experiences, staging surprises, new thoughts and displaced perspectives that lead to alternative modes of thinking about the space between art and science. We invite scientists (physicists, historians, mathematicians, medical students), artists (dancers, musicians, painters, poets, chefs) and not least architects, who reside in these borderlands and wish to share their vision and work. The key intention is to celebrate both art and science as key knowledge building devices.
|
|
3. |
- Apelgren, Peter, et al.
(författare)
-
Chondrocytes and stem cells in 3D-bioprinted structures create human cartilage in vivo.
- 2017
-
Ingår i: PloS one. - : Public Library of Science (PLoS). - 1932-6203. ; 12:12
-
Tidskriftsartikel (refereegranskat)abstract
- Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D)-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm) were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority had retained their macroscopic grid appearance. Constructs consisting of human nasal chondrocytes showed good proliferation ability, with 17.2% of the surface areas covered with proliferating chondrocytes after 60 days. In constructs comprising a mixture of chondrocytes and stem cells, an additional proliferative effect was observed involving chondrocyte production of glycosaminoglycans and type 2 collagen. This clinically highly relevant study revealed 3D bioprinting as a promising technology for the creation of human cartilage.
|
|
4. |
|
|
5. |
- Nygren, Håkan, 1952, et al.
(författare)
-
Mineralization at Titanium Surfaces is a Two-Step Process.
- 2016
-
Ingår i: Journal of functional biomaterials. - : MDPI AG. - 2079-4983. ; 7:1
-
Tidskriftsartikel (refereegranskat)abstract
- Mapping the initial reaction of implants with blood or cell culture medium is important for the understanding of the healing process in bone. In the present study, the formation of low crystalline carbonated hydroxyapatite (CHA) onto commercially pure titanium (Ti) implants from cell culture medium and blood, is described as an early event in bone healing at implants. The Ti-implants were incubated with cell culture medium (DMEM) or whole blood and the surface concentration of Ca, P and HA was analyzed by XPS, EDX and Tof-SIMS. After incubation with DMEM for 16 h and 72 h, EDX and XPS analysis showed stable levels of Ca and P on the Ti-surface. ESEM images showed an even distribution of Ca and P. Further analysis of the XPS results indicated that CHA was formed at the implants. Analysis with ToF-SIMS yielded high m.w. fragments of HA, such as Ca₂PO4 at m/z 174.9 and Ca₃PO₅ at m/z 230.8, as secondary ions at the Ti-surfaces. Analysis of implants incubated in blood for 16 h, with ToF-SIMS, showed initial formation of CHA yielding CaOH as secondary ion. The results indicate that early mineralization at Ti-surfaces is an important step in the healing of implants into bone.
|
|
6. |
- Andersson, Marlene, et al.
(författare)
-
Biomimetic spinning of artificial spider silk from a chimeric minispidroin
- 2017
-
Ingår i: Nature Chemical Biology. - : Springer Science and Business Media LLC. - 1552-4450 .- 1552-4469. ; 254
-
Tidskriftsartikel (refereegranskat)abstract
- Herein we present a chimeric recombinant spider silk protein (spidroin) whose aqueous solubility equals that of native spider silk dope and a spinning device that is based solely on aqueous buffers, shear forces and lowered pH. The process recapitulates the complex molecular mechanisms that dictate native spider silk spinning and is highly efficient; spidroin from one liter of bacterial shake-flask culture is enough to spin a kilometer of the hitherto toughest as-spun artificial spider silk fiber.
|
|
7. |
- Malmberg, Per, 1974, et al.
(författare)
-
Formation of hydroxyapatite on titanium implants in vivo precedes bone-formation during healing
- 2017
-
Ingår i: Biointerphases. - : American Vacuum Society. - 1934-8630 .- 1559-4106. ; 12:4
-
Tidskriftsartikel (refereegranskat)abstract
- The bone material interface has been an area of intense study over many decades, where studies of the healing process ranging from simple mineral deposition in vitro to actual healing in vivo have given important clues to the importance of calcium minerals in the bone/implant interface. Here, the authors use a combination of in vitro cell culture methods and in vivo implantation to study how the role of the spontaneously formed hydroxyapatite layer on Ti-implants for the in vivo-healing into the bone tissue of rat tibia. Initial experiments were made in reduced systems by incubation of TiO2 in cell culture medium and analysis by time of flight secondary ion mass spectrometry (ToF-SIMS) and energy-dispersive x-ray spectroscopy followed by subsequent exposure of human embryological stem cells analyzed by von Kossa staining and environmental scanning electron microsopy. In vivo studies of the bone-material interface was analyzed by ToF-SIMS depth profiling using both C-60(+) ions as well as a gas cluster ion source beam, Ar-1500(+) as sputter source. The low ion yield of the Ar-1500(+) for inorganics allowed the inorganic/organic interface of the implant to be studied avoiding the erosion of the inorganic materials caused by the conventional C-60(+) beam. (C) 2017 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
|
8. |
- Asif, Sana, et al.
(författare)
-
Heparinization of cell surfaces with short peptide-conjugated PEG-lipid regulates thromboinflammation in transplantation of human MSCs and hepatocytes
- 2016
-
Ingår i: Acta Biomaterialia. - : Elsevier BV. - 1742-7061 .- 1878-7568. ; 35, s. 194-205
-
Tidskriftsartikel (refereegranskat)abstract
- Infusion of therapeutic cells into humans is associated with immune responses, including thromboinflammation, which result in a large loss of transplanted cells\ To address these problems, heparinization of the cell surfaces was achieved by a cell-surface modification technique using polyethylene glycol conjugated phospholipid (PEG-lipid) derivatives. A short heparin-binding peptide was conjugated to the PEG-lipid for immobilization of heparin conjugates on the surface of human mesenchymal stem cells (hMSCs) and human hepatocytes. Here three kinds of heparin-binding peptides were used for immobilizing heparin conjugates and examined for the antithrombogenic effects on the cell surface. The heparinized cells were incubated in human whole blood to evaluate their hemocompatibility by measuring blood parameters such as platelet count, coagulation markers, complement markers, and Factor Xa activity. We found that one of the heparin-binding peptides did not show cytotoxicity after the immobilization with heparin conjugates. The degree of binding of the heparin conjugates on the cell surface (analyzed by flow cytometer) depended on the ratio of the active peptide to control peptide. For both human MSCs and hepatocytes in whole-blood experiments, no platelet aggregation was seen in the heparin conjugate-immobilized cell group vs. the controls (non-coated cells or control peptide). Also, the levels of thrombin-antithrombin complex (TAT), C3a, and sC5b-9 were significantly lower than those of the controls, indicating a lower activation of coagulation and complement. Factor Xa analysis indicated that the heparin conjugate was still active on the cell surface at 24 h post-coating. It is possible to immobilize heparin conjugates onto hMSC and human hepatocyte surfaces and thereby protect the cell surfaces from damaging thromboinflammation. Statement of Signigficance We present a promising approach to enhance the biocompatibility of therapeutic cells. Here we used short peptide-conjugated PEG-lipid for cell surface modification and heparin conjugates for the coating of human hepatocytes and MSCs. We screened the short peptides to find higher affinity for heparinization of cell surface and performed hemocompatibility assay of heparinized human hepatocytes and human MSCs in human whole blood. Using heparin-binding peptide with higher affinity, not only coagulation activation but also complement activation was significantly suppressed. Thus, it was possible to protect human hepatocytes and human MSCs from the attack of thromboinflammatory activation, which can contribute to the improvement graft survival. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
|
|
9. |
- Pujari-Palmer, Michael, et al.
(författare)
-
A Novel Class of Injectable Bioceramics That Glue Tissues and Biomaterials
- 2018
-
Ingår i: Materials. - : MDPI AG. - 1996-1944. ; 11:12
-
Tidskriftsartikel (refereegranskat)abstract
- Calcium phosphate cements (CPCs) are clinically effective void fillers that are capable of bridging calcified tissue defects and facilitating regeneration. However, CPCs are completely synthetic/inorganic, unlike the calcium phosphate that is found in calcified tissues, and they lack an architectural organization, controlled assembly mechanisms, and have moderate biomechanical strength, which limits their clinical effectiveness. Herein, we describe a new class of bioinspired CPCs that can glue tissues together and bond tissues to metallic and polymeric biomaterials. Surprisingly, alpha tricalcium phosphate cements that are modified with simple phosphorylated amino acid monomers of phosphoserine (PM-CPCs) bond tissues up to 40-fold stronger (2.5-4 MPa) than commercial cyanoacrylates (0.1 MPa), and 100-fold stronger than surgical fibrin glue (0.04 MPa), when cured in wet-field conditions. In addition to adhesion, phosphoserine creates other novel properties in bioceramics, including a nanoscale organic/inorganic composite microstructure, and templating of nanoscale amorphous calcium phosphate nucleation. PM-CPCs are made of the biocompatible precursors calcium, phosphate, and amino acid, and these represent the first amorphous nano-ceramic composites that are stable in liquids.
|
|
10. |
- Wickham, Abeni, et al.
(författare)
-
Near-Infrared Emitting and Pro-Angiogenic Electrospun Conjugated Polymer Scaffold for Optical Biomaterial Tracking
- 2015
-
Ingår i: Advanced Functional Materials. - : Wiley: 12 months. - 1616-301X .- 1616-3028. ; 25:27, s. 4274-4281
-
Tidskriftsartikel (refereegranskat)abstract
- Noninvasive tracking of biomaterials is vital for determining the fate and degradation of an implant in vivo, and to show its role in tissue regeneration. Current biomaterials have no inherent capacity to enable tracing but require labeling with, for example, fluorescent dyes, or nanoparticles. Here a novel biocompatible fully conjugated electrospun scaffold is described, based on a semiconducting luminescent polymer that can be visualized in situ after implantation using fluorescence imaging. The polymer, poly [2,3-bis-(3-octyloxyphenyl)quinoxaline-5,8-diyl-alt -thiophene-2,5-diyl] (TQ1), is electrospun to form a fibrous mat. The fibers display fluorescence emission in the near-infrared region with lifetimes in the sub-nanosecond range, optimal for in situ imaging. The material shows no cytotoxic behaviors for embryonic chicken cardiomyocytes and mouse myoblasts, and cells migrate onto the TQ1 fibers even in the presence of a collagen substrate. Subcutaneous implantations of the material in rats show incorporation of the TQ1 fibers within the tissue, with limited inflammation and a preponderance of small capillaries around the fibers. The fluorescent properties of the TQ1 fibers are fully retained for up to 90 d following implantation and they can be clearly visualized in tissue using fluorescence and lifetime imaging, thus making it both a pro-angiogenic and traceable biomaterial.
|
|
11. |
- Apelgren, Peter, et al.
(författare)
-
In Vivo Human Cartilage Formation in Three-Dimensional Bioprinted Constructs with a Novel Bacterial Nanocellulose Bioink
- 2019
-
Ingår i: Acs Biomaterials Science & Engineering. - : American Chemical Society (ACS). - 2373-9878. ; 5:5, s. 2482-2490
-
Tidskriftsartikel (refereegranskat)abstract
- Bacterial nanocellulose (BNC) is a 3D network of nanofibrils exhibiting excellent biocompatibility. Here, we present the aqueous counter collision (ACC) method of BNC disassembly to create bioink with suitable properties for cartilage-specific 3D-bioprinting. BNC was disentangled by ACC, and fibril characteristics were analyzed. Bioink printing fidelity and shear-thinning properties were evaluated. Cell-laden bioprinted grid constructs (5 X 5 X 1 mm(3)) containing human nasal chondrocytes (10 M mL(-1)) were implanted in nude mice and explanted after 30 and 60 days. Both ACC and hydrolysis resulted in significantly reduced fiber lengths, with ACC resulting in longer fibrils and fewer negative charges relative to hydrolysis. Moreover, ACC-BNC bioink showed outstanding printability, postprinting mechanical stability, and structural integrity. In vivo, cell-laden structures were rapidly integrated, maintained structural integrity, and showed chondrocyte proliferation, with 32.8 +/- 13.8 cells per mm(2) observed after 30 days and 85.6 +/- 30.0 cells per mm(2) at day 60 (p = 0.002). Furthermore, a full-thickness skin graft was attached and integrated completely on top of the 3D-bioprinted construct. The novel ACC disentanglement technique makes BNC biomaterial highly suitable for 3D-bioprinting and clinical translation, suggesting cell-laden 3D-bioprinted ACC-BNC as a promising solution for cartilage repair.
|
|
12. |
- Engberg, Anna E., et al.
(författare)
-
Prediction of inflammatory responses induced by biomaterials in contact with human blood using protein fingerprint from plasma
- 2015
-
Ingår i: Biomaterials. - : Elsevier BV. - 0142-9612 .- 1878-5905. ; 36, s. 55-65
-
Tidskriftsartikel (refereegranskat)abstract
- Inappropriate complement activation is often responsible for incompatibility reactions that occur when biomaterials are used. Complement activation is therefore a criterion included in legislation regarding biomaterials testing. However, no consensus is yet available regarding appropriate complement-activation-related test parameters. We examined protein adsorption in plasma and complement activation/cytokine release in whole blood incubated with well-characterized polymers. Strong correlations were found between the ratio of C4 to its inhibitor C4BP and generation of 10 (mainly pro-inflammatory) cytokines, including IL-17, IFN-gamma, and IL-6. The levels of complement activation products correlated weakly (C3a) or not at all (C5a, sC5b-9), confirming their poor predictive values. We have demonstrated a direct correlation between downstream biological effects and the proteins initially adhering to an artificial surface after contact with blood. Consequently, we propose the C4/C4BP ratio as a robust, predictor of biocompatibility with superior specificity and sensitivity over the current gold standard. (C) 2014 Elsevier Ltd. All rights reserved.
|
|
13. |
- Gatenholm, Paul, 1956, et al.
(författare)
-
Effect of cultivation conditions on the structure and morphological properties of BNC biomaterials with a focus on vascular grafts
- 2016
-
Ingår i: Bacterial NanoCellulose: A Sophisticated Multifunctional Material. - Boca Raton : CRC Press. - 9781439869925 ; , s. 19-42
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- 20 New materials that are not thrombogenic and have mechanical properties that mimic the native blood vessel are in very great demand. Nanocellulose produced by the bacteria Gluconacetobacter xylinus is a biomaterial that has gained interest in the field of tissue engineering because of its unique properties, such as great mechanical strength, high water content (around 99%), and the ability to be shaped into three-dimensional structures during biosynthesis. The fabrication process of bacterial nanocellulose (BNC) vascular grafts is very unique because the material synthesis and product formation takes place simultaneously. The bio mechanical performance, which includes rupture pressure and compliance along with biological response (endothelialization, blood compatibility, etc.), is dependent on the morphology of a fibrillar network. The network formation is affected by cellulose assembly and bacteria motion, proliferation rate, and other factors. An understanding of the effects of cultivation conditions on BNC network formation is therefore of great importance.
|
|
14. |
- Karlsson, Johan, 1984, et al.
(författare)
-
Stem cell homing using local delivery of plerixafor and stromal derived growth factor-1alpha for improved bone regeneration around Ti-implants
- 2016
-
Ingår i: Journal of Biomedical Materials Research - Part A. - : Wiley. - 1552-4965 .- 1549-3296. ; 104:10, s. 2466-2475
-
Tidskriftsartikel (refereegranskat)abstract
- Triggering of the early healing events, including the recruitment of progenitor cells, has been suggested to promote bone regeneration. In implantology, local drug release technologies could provide an attractive approach to promote tissue regeneration. In this study, we targeted the chemotactic SDF-1a/CXCR4 axis that is responsible e.g. for the homing of stem cells to trauma sites. This was achieved by local delivery of plerixafor, an antagonist to CXCR4, and/or SDF-1a from titanium implants coated with mesoporous titania thin films with a pore size of 7.5 nm. In vitro drug delivery experiments demonstrated that the mesoporous coating provided a high drug loading capacity and controlled release. The subsequent in vivo study in rat tibia showed beneficial effects with respect to bone-implant anchorage and bone-formation along the surface of the implants when plerixafor and SDF-1a were delivered locally. The effect was most prominent by the finding that the combination of the drugs significantly improved the mechanical bone anchorage. These observations suggest that titanium implants with local delivery of drugs for enhanced local recruitment of progenitor cells have the ability to promote osseointegration. This approach may provide a potential strategy for the development of novel implant treatments.
|
|
15. |
- Shah, Furqan A., et al.
(författare)
-
Bioactive glass and glass-ceramic scaffolds for bone tissue engineering
- 2018
-
Ingår i: Bioactive Glasses (Second Edition). - 9780081009369 ; , s. 201-33
-
Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Bioactive glasses and glass-ceramics are a diverse group of materials possessing a unique set of physicochemical properties that make them useful for bone repair. Scaffolds for bone tissue engineering are subject to many requirements including biocompatibility, osteogenesis, biodegradability, and mechanical competence, all of which must be considered in the design features. This chapter addresses various scaffold fabrication techniques for melt-derived and sol-gel-derived compositions, polymer-based organic-inorganic composites, calcium phosphate-based inorganic-inorganic composites, bioactive bone cements, scaffolds based on glass compositions containing specific therapeutic ions, and hybrid materials where the organic and inorganic phases interact at the molecular level. The most important achievements, challenges and potential solutions, as well as the most promising areas of future research involving bioactive glasses and glass-ceramics for bone tissue engineering are presented.
|
|
16. |
- Hedlund, Artur, 1984
(författare)
-
Coagulation of Cellulose: from Ionic-Liquid Solution to Cellulose Nanostructure
- 2018
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Abstract A linear chain of glucose monomers, cellulose, provides the structural reinforcement of the cell walls of plants and constitutes almost half of their dry mass. Wood and other plant-based raw materials are processed on a large industrial scale to isolate the cellulose, which is then dissolved. The resulting solutions can be shaped into films or fibers and solidified as such by immersion in a nonsolvent. The properties of the solidified cellulose products can, however, vary and are frequently not quite satisfactory. In the solidification process, cellulose forms one phase and the nonsolvent and solvent form a second phase, which is later removed through washing and drying. However, these phase separations of ternary mixtures are more complicated than the sentence above indicates. In fact, the details left out decide the properties of those variable materials. This thesis reports on the interdependencies between several parameters and aspects that are critical to cellulose phase separations: compound properties, phase equilibria for the ternary mixtures, the diffusion processes, and the nanostructures formed. Several new experimental methods were developed to measure the critical amounts of nonsolvent that induce coagulation, the mass transport of solvent and nonsolvent, and the rates of coagulation. The cellulose solutions of an ionic liquid, 1ethyl-3methyl-imidazolium acetate, [C2mim][OAc], with varied amounts of a cosolvent, DMSO, were coagulated in water, ethanol (EtOH), or 2-propanol (2PrOH). It was found that 2PrOH is, expressed in molar ratio, the strongest nonsolvent (> EtOH > water). However, the diffusive rates, D, and coagulation rates were in the opposite order (water > EtOH > 2PrOH). The observed differences between nonsolvent compounds were much larger for D[C2mim][OAc] than for DNonSolvent , for the rates of coagulation or for DDMSO, particularly with high cellulose concentration. More differences between water and alcohol as the nonsolvent were observed in the cellulose structures formed. Coagulation in water produced relatively well-ordered crystalline structures, whereas coagulation in alcohol did not. The differences between water and alcohol as the nonsolvent can be explained by different modes of phase separation and differences in nonsolvent interactions with [C2mim][OAc] and cellulose. To show the reader how we arrived at those conclusions, which have not been found in previous literature in the cellulose field, a substantial background regarding the properties and interactions of the compounds is supplied. Networks of cellulose nanofibrils were formed in all the nonsolvents tested, which explained the generally high diffusivities observed and the minor effect of cellulose on diffusion. It appeared that diffusion through the cellulose nanofibril network is similar to diffusion in a mixture of [C2mim][OAc] and nonsolvent only, which was confirmed with a simplistic computer model.
|
|
17. |
- Aronsson, Christopher
(författare)
-
Tunable and modular assembly of polypeptides and polypeptide-hybrid biomaterials
- 2016
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Biomaterials are materials that are specifically designed to be in contact with biological systems and have for a long time been used in medicine. Examples of biomaterials range from sophisticated prostheses used for replacing outworn body parts to ordinary contact lenses. Currently it is possible to create biomaterials that can e.g. specifically interact with cells or respond to certain stimuli. Peptides, the shorter version of proteins, are excellent molecules for fabrication of such biomaterials. By following and developing design rules it is possible to obtain peptides that can self-assemble into well-defined nanostructures and biomaterials.The aim of this thesis is to create ”smart” and tunable biomaterials by molecular self-assembly using dimerizing –helical polypeptides. Two different, but structurally related, polypeptide-systems have been used in this thesis. The EKIV-polypeptide system was developed in this thesis and consists of four 28-residue polypeptides that can be mixed-and-matched to self-assemble into four different coiled coil heterodimers. The dissociation constant of the different heterodimers range from μM to < nM. Due to the large difference in affinities, the polypeptides are prone to thermodynamic social self-sorting. The JR-polypeptide system, on the other hand, consists of several 42-residue de novo designed helix-loop-helix polypeptides that can dimerize into four-helix bundles. In this work, primarily the glutamic acid-rich polypeptide JR2E has been explored as a component in supramolecular materials. Dimerization was induced by exposing the polypeptide to either Zn2+, acidic conditions or the complementary polypeptide JR2K.By conjugating JR2E to hyaluronic acid and the EKIV-polypeptides to star-shaped poly(ethylene glycol), respectively, highly tunable hydrogels that can be self-assembled in a modular fashion have been created. In addition, self-assembly of spherical superstructures has been investigated and were obtained by linking two thiol-modified JR2E polypeptides via a disulfide bridge in the loop region. The thesis also demonstrates that the polypeptides and the polypeptide-hybrids can be used for encapsulation and release of molecules and nanoparticles. In addition, some of the hydrogels have been explored for 3D cell culture. By using supramolecular interactions combined with bio-orthogonal covalent crosslinking reactions, hydrogels were obtained that enabled facile encapsulation of cells that retained high viability.The results of the work presented in this thesis show that dimerizing α–helical polypeptides can be used to create modular biomaterials with properties that can be tuned by specific molecular interactions. The modularity and the tunable properties of these smart biomaterials are conceptually very interesting andmake them useful in many emerging biomedical applications, such as 3D cell culture, cell therapy, and drug delivery.
|
|
18. |
- Huang, Shan, et al.
(författare)
-
Reciprocal relationship between contact and complement system activation on artificial polymers exposed to whole human blood.
- 2016
-
Ingår i: Biomaterials. - : Elsevier. - 0142-9612 .- 1878-5905. ; 77, s. 111-119
-
Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Inappropriate and uncontrolled activation of the cascade systems in the blood is a driving force in adverse inflammatory and thrombotic reactions elicited by biomaterials, but limited data are available on the activation of the contact system by polymers and the present study was undertaken to investigate these mechanisms in established models.METHODS: Polymer particles were incubated in (1) EDTA-plasma (10 mM) to monitor the adsorption of 20 selected proteins; (2) lepirudin-anticoagulated plasma to evaluate contact system activation, monitored by the formation of complexes between the generated proteases factor[F]XIIa, FXIa and kallikrein and the serpins C1-inhibitor [C1INH] and antithrombin [AT]; (3) lepirudin-anticoagulated whole blood to determine cytokine release.RESULTS: Strong negative correlations were found between 10 cytokines and the ratio of deposited FXII/C1INH, generated FXIIa-C1INH complexes, and kallikrein-C1INH complexes. Formation of FXIIa-C1INH complexes correlated negatively with the amount of C3a and positively with deposited IgG.CONCLUSIONS: A reciprocal relationship was found between activation of the contact system and the complement system induced by the polymers studied here. The ratios of FXII/C1INH or C4/C4BP, adsorbed from EDTA-plasma are useful surrogate markers for cytokine release and inflammatory response to materials intended for blood contact.
|
|
19. |
|
|
20. |
- Karlsson, Johan, et al.
(författare)
-
The effect of alendronate on biomineralization at the bone/implant interface.
- 2016
-
Ingår i: Journal of Biomedical Materials Research. Part A. - : Wiley. - 1549-3296 .- 1552-4965. ; 104:3, s. 620-629
-
Tidskriftsartikel (refereegranskat)abstract
- A recent approach to improve the osseointegration of implants is to utilize local drug administration. The presence of an osteoporosis drug may influence both bone quantity and quality at the bone/implant interface. Despite this, the performance of bone-anchoring implants is traditionally evaluated only by quantitative measurements. In the present study, the osteoporosis drug alendronate (ALN) was administrated from mesoporous titania thin films that were coated onto titanium implants. The effect that the drug had on biomineralization was explored both in vitro using simulated body fluid (SBF) and in vivo in a rat tibia model. The SBF study showed that the apatite formation was completely hindered at a high concentration of ALN (0.1 mg/mL). However, when ALN was administrated from the mesoporous coating the surface became completely covered with apatite. Ex vivo characterization of the bone/implant interface using Raman spectroscopy demonstrated that the presence of ALN enhanced the bone mineralization, and that the chemical signature of newly formed bone in the presence of ALN had a higher resemblance to the pre-existing mature bone than to the bone formed without drug. Taken together, this study demonstrates the importance of evaluating the quality of the formed bone to better understand the performance of implants. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A 104A: 620-629, 2016.
|
|
21. |
- Sukhovey, Yurij G., et al.
(författare)
-
Difference between the biologic and chronologic age as an individualized indicator for the skin care intensity selection : skin topography and immune system state studies, parameter correlations with age difference
- 2019
-
Ingår i: Biomedical Dermatology. - : Springer Nature. - 2398-8460. ; 3
-
Tidskriftsartikel (refereegranskat)abstract
- Background: Present research addresses the issue of skin aging and corresponding skin treatment individualization. Particular research question was on the developing of simplified criterion supporting patient-specific decision on the necessity and intensity of skin treatment. Basing on the published results and a wide pool of experimental data, we have formulated a hypothesis that a difference between biologic and chronologic age can be used as an express criterion of skin aging.Methods: In present paper, we report the results of studies with 80 volunteers between 15 and 65 years of age, linking parameters reflecting immune state, skin state, and topography to the difference between biologic and chronologic age. Facial skin topography, skin moisture, sebum level, and skin elasticity were studied using commercial devices. Blood immunology studies were performed using venous blood samples. Correlations between all measured parameters and age difference were calculated. Also, cross correlations between skin cell profile and blood immune profile parameters, and skin roughness parameters were calculated.Results: Age dependencies of the blood immunological parameters on the biologic and chronologic age difference are less pronounced as compared to the changes in skin cell profile parameters. However, the changes in the tendencies when biologic age becomes equal to chronologic one are visible for all studied parameters.All measured skin roughness parameters show correlations with age difference, but average skin roughness and depth of the deepest profile valley have the largest correlation coefficient values. Many of the measured skin cell profile and blood immunology parameters show strong correlations with average skin roughness and deepest profile valley, with some of the coefficients exceeding 0.5–0.6.Conclusions: Basing on own experiments and published research results, it is possible to suggest using the difference between calculated biologic age and chronologic age as an individualized criterion supporting decisions on skin treatment strategy. Further research involving larger numbers of participants and aiming on optimizing the expressions for calculating biologic age could lead to reliable and easily available express criterion supporting the decision making for an individualized skin treatment.
|
|
22. |
- Martinez Avila, Hector, 1985
(författare)
-
Biofabrication, Biomechanics and Biocompatibility of Nanocellulose-based Scaffolds for Auricular Cartilage Regeneration
- 2015
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- In about 2:10,000 births the external part of the ear, the auricle, is severely malformed or absent. Furthermore, tumors and trauma can cause defects to the auricle. For patients with dysplasia of the auricle, and especially for children, an inconspicuous outer appearance with life-like auricles is important for their psychological and emotional well being as well as their psycho-social development. Auricular reconstruction remains a great challenge due to the complexity of surgical reconstruction using rib cartilage. Despite the advances in stem cell technology and biomaterials, auricular cartilage tissue engineering (TE) is still in an early stage of development due to critical requirements demanding appropriate mechanical properties and shape stability of the tissue-engineered construct. This thesis has focused on developing patient-specific tissue-engineered auricles for one-step surgery using a novel biomaterial, bacterial nanocellulose (BNC), seeded with human nasoseptal chondrocytes (hNC) and bone marrow mononuclear cells (MNC).Biomechanical properties of human auricle cartilage were measured and used as a benchmark for tuning BNC properties. In order to meet the biomechanical requirements, a scaffold with bilayer architecture composed of a dense BNC support layer and a macroporous structure was designed. Firstly, the biocompatibility of the dense BNC layer was investigated, demonstrating a minimal foreign body response according to standards set forth in ISO 10993. Secondly, different methods to create macroporous BNC scaffolds were studied and the redifferentiation capacity of hNCs was evaluated in vitro; revealing that macroporous BNC scaffolds support cell ingrowth, proliferation and neocartilage formation. The bilayer BNC scaffold was biofabricated and tested for endotoxins and cytotoxicity before evaluating in long-term 3D culture, and subsequently in vivo for eight weeks—in an immunocompromised animal model. The results demonstrated that the non-pyrogenic and non- cytotoxic bilayer BNC scaffold offers a good mechanical stability and maintains a structural integrity, while providing a porous 3D environment that is suitable for hNCs and MNCs to produce neocartilage, in vitro and in vivo. Furthermore, patient-specific auricular BNC scaffolds with bilayer architecture were biofabricated and seeded with autologous rabbit auricular chondrocytes (rAC) for implantation in an immunocompetent rabbit model for six weeks. The results demonstrated the shape stability of the rAC-seeded scaffolds and neocartilage depositions in the immunocompetent autologous grafts. 3D bioprinting was also evaluated for biofabrication of patient-specific, chondrocyte-laden auricular constructs using a bioink composed of nanofibrillated cellulose and alginate. Bioprinted auricular constructs showed an excellent shape and size stability after in vitro culture. Moreover, this bioink supports redifferentiation of hNCs while offering excellent printability, making this a promising approach for auricular cartilage TE. Furthermore, the use of bioreactors is essential for the development of tissue-engineered cartilage in vitro. Thus, a compression bioreactor was utilized to apply dynamic mechanical stimulation to cell-seeded constructs as a means to enhance production of extracellular matrix in vitro.In this work, a potential clinical therapy for auricular reconstruction using tissue-engineered auricles is demonstrated; where BNC is proposed as a promising non-degradable biomaterial with good chemical and mechanical stability for auricular cartilage TE. Although the primary focus of this thesis is on auricular reconstruction, the methods developed are also applicable in the regeneration of other cartilage tissues such as those found in the nose, trachea, spine and articular joints.
|
|
23. |
- Andersson, Johanna, 1984, et al.
(författare)
-
Stick–slip motion and controlled filling speed by the geometric design of soft micro-channels
- 2018
-
Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 1095-7103 .- 0021-9797. ; 524, s. 139-147
-
Tidskriftsartikel (refereegranskat)abstract
- Hypothesis Liquid can move by capillary action through interconnected porous materials, as in fabric or paper towels. Today mass transport is controlled by chemical modification. It is, however, possible to direct mass transport by geometrical modifications. It is here proposed that it is possible to tailor capillary flow speed in a model system of micro-channels by the angle, size and position of attached side channels. Experiments A flexible, rapid, and cost-effective method is used to produce micro-channels in gels. It involves 3D-printed moulds in which gels are cast. Open channels of micrometre size with several side channels on either one or two sides are produced with tilting angles of 10 – 170°. On a horizontal plane the meniscus of water driven by surface tension is tracked in the main channel. Findings The presence of side channels on one side slowed down the speed of the meniscus in the main channel least. Channels having side channels on both sides with tilting angles of up to 30° indicated tremendously slower flow, and the liquid exhibited a stick-slip motion. Broader side channels decreased the speed more than thinner ones, as suggested by the hypothesis. Inertial forces are suggested to be important in branched channel systems studied here.
|
|
24. |
- Asif, Sana, M.D, PhD student, et al.
(författare)
-
Validation of an MPC polymer coating to attenuate surface- induced cross-talk between the complement and coagulation systems in whole blood in in vitro and in vivo models
- 2019
-
Ingår i: Macromolecular Bioscience. - : Wiley-VCH Verlagsgesellschaft. - 1616-5187 .- 1616-5195. ; 19:5
-
Tidskriftsartikel (refereegranskat)abstract
- Artificial surfaces that come into contact with blood induce an immediate activation of the cascade systems of the blood, leading to a thrombotic and/or inflammatory response that can eventually cause damage to the biomaterial or the patient, or to both. Heparin coating has been used to improve hemocompatibility, and another approach is 2-methacryloyloxyethyl phosphorylcholine (MPC)-based polymer coatings. Here, the aim is to evaluate the hemocompatibility of MPC polymer coating by studying the interactions with coagulation and complement systems using human blood in vitro model and pig in vivo model. The stability of the coatings is investigated in vitro and MPC polymer-coated catheters are tested in vivo by insertion into the external jugular vein of pigs to monitor the catheters' antithrombotic properties. There is no significant activation of platelets or of the coagulation and complement systems in the MPC polymer-coated one, which was superior in hemocompatibility to non-coated matrix surfaces. The protective effect of the MPC polymer coat does not decline after incubation in human plasma for up to 2 weeks. With MPC polymer-coated catheters, it is possible to easily draw blood from pig for 4 days in contrast to the case for non-coated catheters, in which substantial clotting is seen.
|
|
25. |
- Shah, Furqan A., et al.
(författare)
-
Ultrastructural evaluation of shrinkage artefacts induced by fixatives and embedding resins on osteocyte processes and pericellular space dimensions
- 2015
-
Ingår i: Journal of Biomedical Materials Research. Part A. - : Wiley. - 1549-3296 .- 1552-4965. ; 103:4, s. 1565-76
-
Tidskriftsartikel (refereegranskat)abstract
- The integrity of the interface between the osteocyte (Ot) process and the canalicular wall was investigated in terms of change in the lateral dimensions of the Ot process in relation to the canalicular width, i.e., widening of the pericellular space. This has been interpreted as shrinkage of the Ot process relative to the canalicular wall during sample preparation stages of fixation, dehydration, and resin embedding. Sprague-Dawley rat tibial cross-sections were prepared for transmission electron microscopy (TEM). Four different fixative preparations: paraformaldehyde (PF), modified Karnovsky's (MK), glutaraldehyde (GRR) with ruthenium red (GRR), and zinc formalin (ZF); and two different embedding resins: LR Gold (LRG) and Epon812 (Epon) were evaluated. It was found that for LRG embedding, formalin-only fixatives (PF and ZF) induced lower shrinkage than GRR-containing fixatives (MK and GRR). In contrast, for Epon embedding, MK showed the highest shrinkage, while no differences were found between the remaining fixatives (PF, ZF, and GRR). All formalin-containing fixatives (MK, PF, and ZF) induced similar shrinkage in both embedding media. The most dramatic difference was for GRR fixation, which in combination with LRG embedding showed ∼62% more shrinkage than with Epon embedding, suggesting that the combination of GRR fixation and LRG embedding synergistically amplifies Ot shrinkage. These differences likely suggest a role of the resin in secondarily influencing the tissue structure following fixation. Further, the work confirms LRG as a poor embedding medium for bone specimens, as it causes large variations in shrinkage depending on fixation. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2014.
|
|
26. |
- Zhang, Ranran, et al.
(författare)
-
Effects of the hierarchical macro/mesoporous structure on the osteoblast-like cell response
- 2018
-
Ingår i: Journal of Biomedical Materials Research. Part A. - : Wiley. - 1549-3296 .- 1552-4965. ; 106:7, s. 1896-1902
-
Tidskriftsartikel (refereegranskat)abstract
- To improve the success of medical devices, implants with strong surface bioactivity are urgently required. Coatings with a macroporous structure produced by micro-arc oxidation possess advantages, such as strong adhesion to substrate and excellent resistance to wear and corrosion. Mesoporous structures contain pores with sizes of 2-50 nm, which can endow the biomaterials with the ability to enhance osteogenesis and to be loaded with diverse drugs. Thus, in this study, we aimed to evaluate the effects of both macroporous and mesoporous structures using a hierarchical macro/mesoporous structure to modify the titanium implant surface. The behaviors of SaOS-2 human osteosarcoma cells on the macro/mesoporous structure, including initial adhesion, proliferation, alkaline phosphatase (ALP) activity, and collagen secretion, were investigated. Cells that attached on the macro/mesoporous surface showed the highest cell numbers and greatest spreading area after incubation for 1, 2, and 4 h compared with the polished smooth substrate and macroporous surface in the presence of fetal bovine serum (FBS). However, in the absence of FBS, cell adhesion on the polished substrate, macroporous structure, and macro/mesoporous structure did not differ significantly. Cell proliferation on the macroporous and macro/mesoporous surfaces increased compared with that on the smooth substrate surface. Furthermore, ALP activity and collagen secretion were enhanced on the macro/mesoporous structure. Our findings provided important insights into the cellular responses to macro/mesoporous structures in the field of implant surface modification.
|
|
27. |
- Borglin, Johan, 1986, et al.
(författare)
-
Peptide Functionalized Gold Nanoparticles as a Stimuli Responsive Contrast Medium in Multiphoton Microscopy
- 2017
-
Ingår i: Nano Letters. - : American Chemical Society (ACS). - 1530-6984 .- 1530-6992. ; 17:3, s. 2102-2108
-
Tidskriftsartikel (refereegranskat)abstract
- There is a need for biochemical contrast mediators with high signal-to-noise ratios enabling noninvasive biomedical sensing, for example, for neural sensing and protein protein interactions, in addition to cancer diagnostics. The translational challenge is to develop a biocompatible approach ensuring high biochemical contrast while avoiding a raise of the background signal. We here present a concept where gold nanoparticles (AuNPs) can be utilized as a stimuli responsive contrast medium by chemically triggering their ability to exhibit multiphoton-induced luminescence (MIL) when performing multiphoton laser scanning microscopy (MPM). Proof-of-principle is demonstrated using peptide-functionalized AuNPs sensitive to zinc ions (Zn2+). Dispersed particles are invisible in the MPM until addition of millimolar concentrations of Zn2+ upon which MIL is enabled through particle aggregation caused by specific peptide interactions and folding. The process can be reversed by removal of the Zn2+ using a chelator, thereby resuspending the AuNPs. In addition, the concept was demonstrated by exposing the particles to matrix metalloproteinase-7 (MMP-7) causing peptide digestion resulting in AuNP aggregation, significantly elevating the MIL signal from the background. The approach is based on the principle that aggregation shifts the plasmon resonance, elevating the absorption cross section in the near-infrared wavelength region enabling onset of MIL. This Letter demonstrates how biochemical sensing can be obtained in far-field MPM and should be further exploited as a future tool for noninvasive optical biosensing.
|
|
28. |
- Lindahl, Carl, et al.
(författare)
-
Biomimetic calcium phosphate coating of additively manufactured porous CoCr implants
- 2015
-
Ingår i: Applied Surface Science. - : Elsevier BV. - 0169-4332 .- 1873-5584. ; 353, s. 40-47
-
Tidskriftsartikel (refereegranskat)abstract
- The aim of this work was to study the feasibility to use a biomimetic method to prepare biomimetic hydroxyapatite (HA) coatings on CoCr substrates with short soaking times and to characterize the properties of such coatings. A second objective was to investigate if the coatings could be applied to porous CoCr implants manufactured by electron beam melting (EBM). The coating was prepared by immersing the pretreated CoCr substrates and EBM implants into the phosphate-buffered solution with Ca2+ in sealed plastic bottles, kept at 60 degrees C for 3 days. The formed coating was partially crystalline, slightly calcium deficient and composed of plate-like crystallites forming roundish flowers in the size range of 300-500 nm. Cross-section imaging showed a thickness of 300-500 nm. In addition, dissolution tests in Tris-HCl up to 28 days showed that a substantial amount of the coating had dissolved, however, undergoing only minor morphological changes. A uniform coating was formed within the porous network of the additive manufactured implants having similar thickness and morphology as for the flat samples. In conclusion, the present coating procedure allows coatings to be formed on CoCr and could be used for complex shaped, porous implants made by additive manufacturing.
|
|
29. |
- Olsson, Rickard, 1959-, et al.
(författare)
-
Production of osseointegrating (bone bonding) surfaces on titanium screws by laser melt disruption
- 2018
-
Ingår i: Journal of Laser Applications. - Melville, NY : Laser Institute of America. - 1042-346X .- 1938-1387. ; 30:4
-
Tidskriftsartikel (refereegranskat)abstract
- Several techniques can be used to modify implant surfaces in order to accelerate bone growth around titanium implants. One method is to generate a surface structure which stimulates bone growth and remodeling. This paper describes and explains a nonablative method for producing osseointegrating (structural and functional bone bonding) surfaces on titanium implants using laser processing. The focus is especially on surface texturing of dental implant screws, where the ability of a Nd:YAG laser to generate "splashy" surfaces covered in resolidified microscale droplets coated with nanoscale surface oxides is assessed. The surfaces produced were analyzed by a scanning electron microscope and energy dispersive x-ray spectroscopy. It is concluded that laser processing using Q-pulsed Nd:YAG lasers can generate surfaces which match the demands set by clinical experience. One important characteristic of the surfaces discussed here is that they involve overhanging features which are suitable for trapping red blood cells and which cannot be created by mechanical or chemical roughening techniques. © 2018 Laser Institute of America.
|
|
30. |
- Alenezi, Ali, et al.
(författare)
-
Osseointegration effects of local release of strontium ranelate from implant surfaces in rats
- 2019
-
Ingår i: Journal of Materials Science: Materials in Medicine. - : Springer Science and Business Media LLC. - 0957-4530 .- 1573-4838. ; 30:10, s. 116-
-
Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND : Numerous studies have reported the beneficial effects of strontium on bone growth, particularly by stimulating osteoblast proliferation and differentiation. Thus, strontium release around implants has been suggested as one possible strategy to enhance implant osseointegration. AIM : This study aimed to evaluate whether the local release of strontium ranelate (Sr-ranelate) from implants coated with mesoporous titania could improve bone formation around implants in an animal model. MATERIALS AND METHODS : Mesoporous titania (MT) thin coatings were formed utilizing the evaporation induced self-assembly (EISA) method using Pluronic (P123) with or without the addition of poly propylene glycol (PPG) to create materials with two different pore sizes. The MT was deposited on disks and mini-screws, both made of cp Ti grade IV. Scanning electron microscopy (SEM) was performed to characterize the MT using a Leo Ultra55 FEG instrument (Zeiss, Oberkochen, Germany). The MT was loaded with Sr-ranelate using soaking and the drug uptake and release kinetics to and from the surfaces were evaluated using quartz crystal microbalance with dissipation monitoring (QCM-D) utilizing a Q-sense E4 instrument. For the in vivo experiment, 24 adult rats were analyzed at two time points of implant healing (2 and 6 weeks). Titanium implants shaped as mini screws were coated with MT films and divided into two groups; supplied with Sr-ranelate (test group) and without Sr-ranelate (control group). Four implants (both test and control) were inserted in the tibia of each rat. The in vivo study was evaluated using histomorphometric analyses of the implant/bone interphase using optical microscopy. RESULTS : SEM images showed the successful formation of evenly distributed MT films covering the entire surface with pore sizes of 6 and 7.2 nm, respectively. The QCM-D analysis revealed an absorption of 3300 ng/cm2 of Sr-ranelate on the 7.2 nm MT, which was about 3 times more than the observed amount on the 6 nm MT (1200 ng/cm2). Both groups showed sustained release of Sr-ranelate from MT coated disks. The histomorphometric analysis revealed no significant differences in bone implant contact (BIC) and bone area (BA) between the implants with Sr-ranelate and implants in the control groups after 2 and 6 weeks of healing (BIC with a p-value of 0.43 after 2 weeks and 0.172 after 6 weeks; BA with a p-value of 0.503 after 2 weeks, and 0.088 after 6 weeks). The mean BIC and BA values within the same group showed significant increase among all groups between 2 and 6 weeks. CONCLUSION : This study could not confirm any positive effects of Sr-ranelate on implant osseointegration.
|
|
31. |
- Raina, Deepak
(författare)
-
Biomaterials as carriers for bone active molecules-An approach to create off-the-shelf bone substitutes
- 2018
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Bone tissue is commonly transplanted during orthopedic surgeries, primarily for the management of bone defects caused by trauma or various orthopedic conditions including, but not limited to, infections and tumours. Bone grafts are a surgeon’s choice, but their associated drawbacks are paving the way for biomaterial based bone graft substitutes. Biomaterials inherently lack the ability to induce significant amount of bone growth due to which they may be combined with cells, biomaterials, growth factors and drugs to regenerate functional bone tissue. This thesis focused on characterizing biomaterial carriers that can locally deliver bone active molecules for bone regeneration and potentially act as an alternative to conventional bone grafting. We focused on the delivery of recombinant human bone morphogenic protein-2 (rhBMP-2) as a bone inducing anabolic growth factor. Simultaneously, we have used an osteoclast inhibiting bisphosphonate, zoledronic acid (ZA) to prevent BMP-2 induced premature bone resorption. Three different biomaterials scaffolds; a microporous calcium sulphate (CaS)/hydroxyapatite (HA), a macroporous gelatin-CaS/HA and a collagen membrane were used in distinct animal models of bone regeneration.The carrier properties of the three biomaterials in the ectopic muscle pouch model (studies 1, 4 and 5) showed that the tested materials were efficient carriers of rhBMP-2 and ZA and that co-delivery of rhBMP-2 and ZA regenerated higher volume of bone compared to rhBMP-2 alone. Studies 2& 3 show that the CaS/HA material locally delivering ZA or ZA+rhBMP-2 could be efficiently used for bone regeneration in clinically relevant bone defect models. These studies also indicated that local delivery of ZA not only has an anti-osteoclast effect but it also has an anabolic role. Study 4 compared the developed porous biomaterial with the current FDA approved collagen sponge and results indicated that the developed biomaterial outperforms the current marketed product for the delivery of rhBMP-2. During this study, it was also established that co-delivery of rhBMP-2 with ZA could reduce the effective rhBMP-2 doses by up to four times, which is crucial to reinstate BMPs into the clinics. Study 5 was a follow-up of study 2 separating the metaphyseal defect healing in two stages; 1) Healing the cancellous bone using a porous material and 2) Guiding cortical regeneration using a thin collagen membrane. Significantly better cortical healing was noted using this approach in comparison to study 2.In summary, this work describes promising strategies for bone regeneration. It established how the release of bone active molecules can be controlled by the choice of carrier material and how we can decrease the minimally effective dose of rhBMP- 2 by up to four times. These findings can potentially be translated from the bench to the bedside. The materials and methods developed within the scope of this work can be used in a variety of orthopedic conditions and can provide the surgeon with an effective off-the-shelf substitute for bone replacement, in turn leading to improved care of the patient.
|
|
32. |
- Shchukarev, Andrey, et al.
(författare)
-
Surface characterization of insulin-coated Ti6Al4V medical implants conditioned in cell culture medium: An XPS study
- 2017
-
Ingår i: Journal of Electron Spectroscopy and Related Phenomena. - : Elsevier BV. - 0368-2048 .- 1873-2526. ; 216, s. 33-38
-
Tidskriftsartikel (refereegranskat)abstract
- © 2017 Elsevier B.V.Surface characterization of insulin-coated Ti6Al4V medical implants, after incubation in α-minimum essential medium (α-MEM), was done by X-ray photoelectron spectroscopy (XPS), in order to analyze the insulin behavior at the implant – α-MEM interface. In the absence of serum proteins in cell culture medium, the coated insulin layer remained intact, but experienced a time-dependent structural transformation exposing hydrophobic parts of the protein toward the solution. The presence of fetal bovine serum (FBS) in the medium resulted in partial substitution of insulin by serum proteins. In spite of some insulin release, the remaining coated layer demonstrated a direct surface effect by stabilizing the structure of protein competitors, and by supporting the accumulation of calcium and phosphate ions at the interface. A structurally stable protein layer with incorporated calcium and phosphate ions at the implant–tissue interface could be an important prerequisite for enhanced bone formation.
|
|
33. |
- Joffre, Thomas, 1987-, et al.
(författare)
-
Characterization of interfacial stress transfer ability in acetylation-treated wood fibre composites using X-ray microtomography
- 2017
-
Ingår i: Industrial crops and products (Print). - : Elsevier BV. - 0926-6690 .- 1872-633X. ; 95, s. 43-49
-
Tidskriftsartikel (refereegranskat)abstract
- The properties of the fibre/matrix interface contribute to stiffness, strength and fracture behaviour of fibre-reinforced composites. In cellulosic composites, the limited affinity between the hydrophilic fibres and the hydrophobic thermoplastic matrix remains a challenge, and the reinforcing capability of the fibres is hence not fully utilized. A direct characterisation of the stress transfer ability through pull-out tests on single fibres is extremely cumbersome due to the small dimension of the wood fibres. Here a novel approach is proposed: the length distribution of the fibres sticking out of the matrix at the fracture surface is approximated using X-ray microtomography and is used as an estimate of the adhesion between the fibres and the matrix. When a crack grows in the material, the fibres will either break or be pulled-out of the matrix depending on their adhesion to the matrix: good adhesion between the fibres and the matrix should result in more fibre breakage and less pull-out of the fibres than poor adhesion. The effect of acetylation on the adhesion between the wood fibres and the PLA matrix was evaluated at different moisture contents using the proposed method. By using an acetylation treatment of the fibres it was possible to improve the strength of the composite samples soaked in the water by more than 30%.
|
|
34. |
- Larsson Wexell, Cecilia, 1965, et al.
(författare)
-
Electropolished titanium implants with a mirror-like surface support osseointegration and bone remodelling
- 2016
-
Ingår i: Advances in Materials Science and Engineering. - : Hindawi Limited. - 1687-8434 .- 1687-8442.
-
Tidskriftsartikel (refereegranskat)abstract
- This work characterises the ultrastructural composition of the interfacial tissue adjacent to electropolished, commercially pure titanium implants with and without subsequent anodisation, and it investigates whether a smooth electropolished surface can support bone formation in a manner similar to surfaces with a considerably thicker surface oxide layer. Screw-shaped implants were electropolished to remove all topographical remnants of the machining process, resulting in a thin spontaneously formed surface oxide layer and a smooth surface. Half of the implants were subsequently anodically oxidised to develop a thickened surface oxide layer and increased surface roughness. Despite substantial differences in the surface physicochemical properties, the microarchitecture and the composition of the newly formed bone were similar for both implant surfaces after 12 weeks of healing in rabbit tibia. A close spatial relationship was observed between osteocyte canaliculi and both implant surfaces. On the ultrastructural level, the merely electropolished surface showed the various stages of bone formation, for example, matrix deposition and mineralisation, entrapment of osteoblasts within the mineralised matrix, and their morphological transformation into osteocytes. The results demonstrate that titanium implants with a mirror-like surface and a thin, spontaneously formed oxide layer are able to support bone formation and remodelling.
|
|
35. |
- Petrou, Georgia, 1991-
(författare)
-
Investigating mucin interactions with diverse surfaces for biomedical applications
- 2019
-
Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Mucous membranes are covered with mucus, a viscoelastic hydrogel that plays an essential role in their protection from shear and pathogens. The viscoelasticity of mucus is owing to mucins, a group of densely glycosylated proteins. Mucins can interact with a wide range of surfaces; thus, there is big interest in exploring and manipulating such interactions for biomedical applications. This thesis presents investigations of mucin interactions with hydrophobic surfaces in order to identify the key features of mucin lubricity, as well as describes the development of materials that are optimized to interact with mucins. In Paper I we investigated the domains which make mucins outstanding boundary lubricants. The results showed that the hydrophobic terminal domains of mucins play a crucial role in the adsorption and lubrication on hydrophobic surfaces. Specifically, protease digestion of porcine gastric mucins and salivary mucins resulted in the cleavage of these domains and the loss of lubricity and surface adsorption. However, a “rescue” strategy was successfully carried out by grafting hydrophobic phenyl groups to the digested mucins and enhancing their lubricity. This strategy also enhanced the lubricity of polymers which are otherwise bad lubricants. In Paper II we developed mucoadhesive materials based on genetically engineered partial spider silk proteins. The partial spider silk protein 4RepCT was successfully functionalized with six lysines (pLys-4RepCT), or the Human Galectin-3 Carbohydrate Recognition Domain (hGal3-4RepCT). These strategies were aiming to either non-specific electrostatic interactions between the positive lysines and the negative mucins, or specific binding between the hGal3 and the mucin glycans. Coatings, fibers, meshes and foams were prepared from the new silk proteins, and the adsorption of porcine gastric mucins and bovine submaxillary mucins was measured, demonstrating enhanced adsorption. The work presented demonstrates how mucin-material interactions can provide us with valuable information for the development of new biomaterials. Specifically, mucin-based and mucin-inspired lubricants could provide desired lubrication to a wide range of surfaces, while our new silk based materials could be valuable tools for the development of mucosal dressings.
|
|
36. |
- Fursatz, Marian, et al.
(författare)
-
Functionalization of bacterial cellulose wound dressings with the antimicrobial peptide ε-poly-L-Lysine
- 2018
-
Ingår i: Biomedical Materials. - : Institute of Physics Publishing (IOPP). - 1748-6041 .- 1748-605X. ; 13
-
Tidskriftsartikel (refereegranskat)abstract
- Wound dressings based on bacterial cellulose (BC) can form a soft and conformable protective layer that can stimulate wound healing while preventing bacteria from entering the wound. Bacteria already present in the wound can, however, thrive in the moist environment created by the BC dressing which can aggravate the healing process. Possibilities to render the BC antimicrobial without affecting the beneficial structural and mechanical properties of the material would hence be highly attractive. Here we present methods for functionalization of BC with ε-Poly-L-Lysine (ε-PLL), a non-toxic biopolymer with broad-spectrum antimicrobial activity. Low molecular weight ε-PLL was cross-linked in pristine BC membranes and to carboxymethyl cellulose (CMC) functionalized BC using carbodiimide chemistry. The functionalization of BC with ε-PLL inhibited growth of S. epidermidis on the membranes but did not affect the cytocompatibility to cultured human fibroblasts as compared to native BC. The functionalization had no significant effects on the nanofibrous structure and mechanical properties of the BC. The possibility to functionalize BC with ε-PLL is a promising, green and versatile approach to improve the performance of BC in wound care and other biomedical applications.
|
|
37. |
- Calon, Tim, et al.
(författare)
-
Multimodal analysis of the tissue response to a bone-anchored hearing implant. : 11.Calon TGA, Johansson ML, Omar O, Shah FA, Budding D, Trobos M, Thomsen P, Stokroos RJ, Palmquist
- 2019
-
Ingår i: 2nd Politzer Society Meeting/2nd World Congress of Otology, 28 May - 1 June, 2019, Warsaw, Poland..
-
Konferensbidrag (övrigt vetenskapligt/konstnärligt)
|
|
38. |
- Wu, Dan, 1990-, et al.
(författare)
-
Young’s modulus of trabecular bone at the tissue level : A review
- 2018
-
Ingår i: Acta Biomaterialia. - : Elsevier BV. - 1742-7061 .- 1878-7568. ; 78, s. 1-12
-
Forskningsöversikt (refereegranskat)abstract
- The tissue-level Young’s modulus of trabecular bone is important for detailed mechanical analysis of bone and bone-implant mechanical interactions. However, the heterogeneity and small size of the trabecular struts complicate an accurate determination. Methods such as micro-mechanical testing of single trabeculae, ultrasonic testing, and nanoindentation have been used to estimate the trabecular Young’s modulus. This review summarizes and classifies the trabecular Young’s moduli reported in the literature. Information on species, anatomic site, and test condition of the samples has also been gathered. Advantages and disadvantages of the different methods together with recent developments are discussed, followed by some suggestions for potential improvement, for future work. In summary, this review provides a thorough introduction to the approaches used for determining trabecular Young’s modulus, highlights important considerations when applying these methods and summarizes the reported Young’s modulus for follow-up studies on trabecular properties.
|
|
39. |
- Barreto Henriksson, Helena, et al.
(författare)
-
Determination of mechanical and rheological properties of a cell-loaded peptide gel during ECM production
- 2019
-
Ingår i: International Journal of Pharmaceutics. - : Elsevier BV. - 0378-5173 .- 1873-3476. ; 563, s. 437-444
-
Tidskriftsartikel (refereegranskat)abstract
- The development of an injectable biomaterial that supports cell survival and maintains or promotes nucleus pulposus (NP) phenotype could aid delivery of cells to degenerated NPs causing low back pain. Mesenchymal cells were loaded and grown in a synthetic peptide gel, PuraMatrix (R). Cells were observed within the gels over 0-28 days, and accumulation of glycosaminoglycans were detected by histological staining. The mechanical properties of the cell-loaded constructs, and the change of the mechanical properties were studied using stress relaxation of the gels under compression and confinement. The PuraMatrix (R) gel was shown to relax fast on compression indicating that the fluid could easily flow out of the gel, and thus indicating the presence of large pores/voids. The presence of these pores/voids was further supported by high mobility of dextran molecules, determined using fluorescence recovery after photo bleaching. The stress required to deform the cell-loaded constructs to a specific strain increases at day 21, at which point the presence of glycosaminoglycans within the cell-loaded constructs was also observed. The results provide evidence of changes in mechanical properties of the PuraMatrix (R) matrix upon excretion of the extracellular matrix by the cells.
|
|
40. |
|
|
41. |
- Bano, Fouzia, et al.
(författare)
-
Single-molecule unbinding forces between the polysaccharide hyaluronan and its binding proteins
- 2018
-
Ingår i: Biophysical Journal. - : Biophysical society. - 0006-3495 .- 1542-0086. ; 114:12, s. 2910-2922
-
Tidskriftsartikel (refereegranskat)abstract
- The extracellular polysaccharide hyaluronan (HA) is ubiquitous in all vertebrate tissues, where its various functions are encoded in the supramolecular complexes and matrices that it forms with HA-binding proteins (hyaladherins). In tissues, these supramolecular architectures are frequently subjected to mechanical stress, yet how this affects the intermolecular bonding is largely unknown. Here, we used a recently developed single-molecule force spectroscopy platform to analyze and compare the mechanical strength of bonds between HA and a panel of hyaladherins from the Link module superfamily, namely the complex of the proteoglycan aggrecan and cartilage link protein, the proteoglycan versican, the inflammation-associated protein TSG-6, the HA receptor for endocytosis (stabilin-2/HARE), and the HA receptor CD44. We find that the resistance to tensile stress for these hyaladherins correlates with the size of the HA-binding domain. The lowest mean rupture forces are observed for members of the type A subgroup (i.e., with the shortest HA-binding domains; TSG-6 and HARE). In contrast, the mechanical stability of the bond formed by aggrecan in complex with cartilage link protein (two members of the type C subgroup, i.e., with the longest HA-binding domains) and HA is equal or even superior to the high affinity streptavidin⋅biotin bond. Implications for the molecular mechanism of unbinding of HA⋅hyaladherin bonds under force are discussed, which underpin the mechanical properties of HA⋅hyaladherin complexes and HA-rich extracellular matrices.
|
|
42. |
- Bano, Fouzia, et al.
(författare)
-
Unraveling the complexity of the interactions of DNA nucleotides with gold by single molecule force spectroscopy
- 2015
-
Ingår i: Nanoscale. - : Royal Society of Chemistry. - 2040-3364 .- 2040-3372. ; 7:46, s. 19528-19533
-
Tidskriftsartikel (refereegranskat)abstract
- Addressing the effect of different environmental factors on the adsorption of DNA to solid supports is critical for the development of robust miniaturized devices for applications ranging from biosensors to next generation molecular technology. Most of the time, thiol-based chemistry is used to anchor DNA on gold – a substrate commonly used in nanotechnology – and little is known about the direct interaction between DNA and gold. So far there have been no systematic studies on the direct adsorption behavior of the deoxyribonucleotides (i.e., a nitrogenous base, a deoxyribose sugar, and a phosphate group) and on the factors that govern the DNA–gold bond strength. Here, using single molecule force spectroscopy, we investigated the interaction of the four individual nucleotides, adenine, guanine, cytosine, and thymine, with gold. Experiments were performed in three salinity conditions and two surface dwell times to reveal the factors that influence nucleotide–Au bond strength. Force data show that, at physiological ionic strength, adenine–Au interactions are stronger, asymmetrical and independent of surface dwell time as compared to cytosine–Au and guanine–Au interactions. We suggest that in these conditions only adenine is able to chemisorb on gold. A decrease of the ionic strength significantly increases the bond strength for all nucleotides. We show that moderate ionic strength along with longer surface dwell period suggest weak chemisorption also for cytosine and guanine.
|
|
43. |
- Lindahl, Carl, et al.
(författare)
-
The influence of Sr content in calcium phosphate coatings
- 2015
-
Ingår i: Materials Science & Engineering C-Materials for Biological Applications. - : Elsevier BV. - 0928-4931 .- 1873-0191. ; 53, s. 322-330
-
Tidskriftsartikel (refereegranskat)abstract
- In this study calcium phosphate coatings with different amounts of strontium.(Sr) were prepared using a biomineralization method. The incorporation of Sr changed the composition and morphology of coatings from plate-like to sphere-like morphology. Dissolution testing indicated that the solubility of the coatings increased with increased Sr concentration. Evaluation of extracts (with Sr concentrations ranging from 0 to 237 mu g/mL) from the HA, 0.06Sr, 0.6Sr, and 12Sr coatings during in vitro cell cultures showed that Sr incorporation into coatings significantly enhanced the ALP activity in comparison to cells treated with control and HA eluted media. These findings show that calcium phosphate coatings could promote osteogenic differentiation even in a low amount of strontium. (C) 2015 Elsevier B.V. All rights reserved.
|
|
44. |
- Lotsari, Antiope, 1981, et al.
(författare)
-
Transformation of amorphous calcium phosphate to bone-like apatite
- 2018
-
Ingår i: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723 .- 2041-1723. ; 9:1
-
Tidskriftsartikel (refereegranskat)abstract
- Mineralisation of calcium phosphates in bone has been proposed to proceed via an initial amorphous precursor phase which transforms into nanocrystalline, carbonated hydroxyapatite. While calcium phosphates have been under intense investigation, the exact steps during the crystallisation of spherical amorphous particles to platelet-like bone apatite are unclear. Herein, we demonstrate a detailed transformation mechanism of amorphous calcium phosphate spherical particles to apatite platelet-like crystals, within the confined nanodomains of a bone-inspired nanocomposite. The transformation is initiated under the presence of humidity, where nanocrystalline areas are formed and crystallisation advances via migration of nanometre sized clusters by forming steps at the growth front. We propose that such transformation is a possible crystallisation mechanism and is characteristic of calcium phosphates from a thermodynamic perspective and might be unrelated to the environment. Our observations provide insight into a crucial but unclear stage in bone mineralisation, the origins of the nanostructured, platelet-like bone apatite crystals.
|
|
45. |
- Olsson, Pär A T, 1981-, et al.
(författare)
-
Ab initio investigation of monoclinic phase stability and martensitic transformation in crystalline polyethylene
- 2018
-
Ingår i: Physical Review Materials. - : American Physical Society. - 2475-9953. ; 2:7, s. 7-13
-
Tidskriftsartikel (refereegranskat)abstract
- We study the phase stability and martensitic transformation of orthorhombic and monoclimic polyethylene by means of density functional theory using the nonempirical consistent-exchange vdW-DF-cx functional [Phys. Rev. B 89, 035412 (2014)]. The results show that the orthorhombic phase is the most stable of the two. Owing to the occurrence of soft librational phonon modes, the monoclimic phase is predicted not to be stable at zero pressure and temperature, but becomes stable when subjected to compressive transverse deformations that pin the chains and prevent them from wiggling freely. This theoretical characterization, or prediction, is consistent with the fact that the monoclimic phase is only observed experimentally when the material is subjected to mechanical loading. Also, the estimated threshold energy for the combination of lattice deformation associated with the T1 and T2 transformation paths (between the orthorhombic and monoclimic phases) and chain shuffling is found to be sufficiently low for thermally activated back transformations to occur. Thus, our prediction is that the crystalline part can transform back from the monoclimc to the orthorhombic phase upon unloading and/or annealing, which is consistent with experimental observations. Finally, we observe how a combination of such phase transformations can lead to a fold-plane reorientation from {110} to {100} type in a single orthorhombic crystal.
|
|
46. |
- Procter, Philip, et al.
(författare)
-
Designing A Commercial Biomaterial For A Specific Unmet Clinical Need – : An Adhesive Odyssey
- 2018
-
Konferensbidrag (refereegranskat)abstract
- There are clinical situations in fracture repair, e.g. osteochondral fragments, where current implant hardware is insufficient. The proposition of an adhesive enabling fixation and healing has been considered but no successful candidate has emerged thus far. The many preclinical and few clinical attempts include fibrin glue, mussel adhesive and even “Kryptonite” (US bone void filler). The most promising recent attempts are based on phosphorylating amino acids, part of a common cellular adhesion mechanism linking mussels, caddis fly larvae, and mammals. Rapid high bond strength development in the wetted fatty environment of fractured bone, that is sustained during biological healing, is challenging to prove both safety and efficacy. Additionally, there are no “predicate” preclinical animal and human models which led the authors to develop novel evaluations for an adhesive candidate “OsStictm” based on calcium salts and amino acids. Adhesive formulations were evaluated in both soft (6/12 weeks) and hard tissue (3,7,10,14 & 42 days) safety studies in murine models. The feasibility of a novel adhesiveness test, initially proven in murine cadaver femoral bone, is being assessed in-vivo (3,7,10,14 & 42 days) in bilateral implantations with a standard tissue glue as the control. In parallel an ex-vivo human bone model using freshly harvested human donor bone is under development to underwrite the eventual clinical application of such an adhesive. This is part of a risk mitigation project bridging between laboratory biomaterial characterisation and a commercial biomaterial development where safety and effectiveness have to meet today´s new medical device requirements.
|
|
47. |
- Andersson, Marlene, et al.
(författare)
-
Biomimetic spinning of artificial spider silk from a chimeric minispidroin
- 2017
-
Ingår i: Nature Chemical Biology. - : Nature Publishing Group. - 1552-4450 .- 1552-4469. ; 13:3, s. 262-
-
Tidskriftsartikel (refereegranskat)abstract
- Herein we present a chimeric recombinant spider silk protein (spidroin) whose aqueous solubility equals that of native spider silk dope and a spinning device that is based solely on aqueous buffers, shear forces and lowered pH. The process recapitulates the complex molecular mechanisms that dictate native spider silk spinning and is highly efficient; spidroin from one liter of bacterial shake-flask culture is enough to spin a kilometer of the hitherto toughest as-spun artificial spider silk fiber.
|
|
48. |
- Ekstrand-Hammarström, Barbro, et al.
(författare)
-
TiO2 nanoparticles tested in a novel screening whole human blood model of toxicity trigger adverse activation of the kallikrein system at low concentrations
- 2015
-
Ingår i: Biomaterials. - : Elsevier BV. - 0142-9612 .- 1878-5905. ; 51, s. 58-68
-
Tidskriftsartikel (refereegranskat)abstract
- There is a compelling need to understand and assess the toxicity of industrially produced nanoparticles (NPs). In order to appreciate the long-term effects of NPs, sensitive human-based screening tests that comprehensively map the NP properties are needed to detect possible toxic mechanisms. Animal models can only be used in a limited number of test applications and are subject to ethical concerns, and the interpretation of experiments in animals is also distorted by the species differences. Here, we present a novel easy-to-perform highly sensitive whole-blood model using fresh non-anticoagulated human blood, which most justly reflects complex biological cross talks in a human system. As a demonstrator of the tests versatility, we evaluated the toxicity of TiO2 NPs that are widely used in various applications and otherwise considered to have relatively low toxic properties. We show that TiO2 NPs at very low concentrations (50 ng/mL) induce strong activation of the contact system, which in this model elicits thromboinflammation. These data are in line with the finding of components of the contact system in the protein corona of the TiO2 NPs after exposure to blood. The contact system activation may lead to both thrombotic reactions and generation of bradykinin, thereby representing fuel for chronic inflammation in vivo and potentially long-term risk of autoimmunity, arteriosclerosis and cancer. These results support the notion that this novel whole-blood model represents an important contribution to testing of NP toxicity. (C) 2015 Elsevier Ltd. All rights reserved.
|
|
49. |
|
|
50. |
- Skyttner, Camilla, 1985-
(författare)
-
Peptide-Liposome Model Systems for Triggered Release
- 2018
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Liposomes are widely used in drug delivery to improve drug efficacy and to reduce side effects. For liposome-encapsulated drugs to become bioavailable and provide a therapeutic effect they must be released, which typically is a slow process that primarily relies on passive diffusion, liposome rupture or endocytotic uptake. Achieving drug concentrations within the therapeutic window can thus be challenging, resulting in poor efficacy and higher risks drug resistance. Finding means to modulate lipid membrane integrity and to trigger rapid and efficient release of liposomal cargo is thus critical to improve current and future liposomal drug delivery systems. The possibilities to tailor lipid composition and surface functionalization is vital for drug delivery applications but also make liposomes attractive model systems for studies of membrane active biomolecules.The overall aim of this thesis work has been to develop new strategies for triggering and controlling changes in lipid membrane integrity and to study the interactions of membrane active peptides with model lipid membranes using both de novo designed and biologically derived synthetic amphipathic cationic peptides. Two different sets of designed peptides have been explored that can fold and heterodimerize into a coiled coil and helix-loop-helix fourhelix bundle, respectively. Conjugation of the cationic lysine rich peptides to liposomes triggered a rapid and concentration dependent release. The additions of their corresponding glutamic acid-rich complementary peptides inhibited the release of liposomal cargo. Possibilities to reduce the inhibitory effect by both proteolytic digestion of the inhibitory peptide and by means of heterodimer exchange have been investigated. Moreover, the effects of peptide size and composition and ability to fold have been studied in order to elucidate the factors that influence the membrane permeabilizing effects of the peptides.In addition, the membrane activity of a the two-peptide bacteriocin PLNC8α and PLNC8β has been explored using liposomes as a model system. PLNC8αβ are expressed by Lactobacillus plantarum and were shown to display pronounced membrane-partition folding coupling, leading to rapid release of liposome encapsulated carboxyfluorescein. PLNC8αβ also kill and suppressed growth of the gram-negative bacteria Porphyromonas gingivalis by efficiently damaging the bacterial membrane.Although membrane active peptides are highly efficient in perturbing lipid membrane integrity, possibilities to trigger release using external stimuli are also of large interest for therapeutic applications. Light-induced heating of liposome encapsulated gold nanoparticles (AuNPs) has been shown by others as a potential strategy to trigger drug release. To facilitate fabrication of thermoplasmonic liposome systems we developed a simple method for synthesis of small AuNPs inside liposomes, using the liposomes as nanoscale reaction vessels.The work presented in this thesis provides new knowledge and techniques for future development of liposome-based drug delivery systems, peptide-based therapeutics and increase our understanding of peptide-lipid interactions.
|
|