Self-Assembly of Recombinant Silk as a Strategy for Chemical-Free Formation of Bioactive Coatings: A Real-Time Study

Nilebäck, Linnea; Hedin, Jesper; Widhe, Mona; Floderus, Lotta S.; Krona, Annika; Bysell, Helena; Hedhammar, My

doi:10.1021/acs.biomac.6b01721

Cited by 52 publications

(121 citation statements)

References 40 publications

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“…When the flow rate is kept this low, the adsorption behavior is considered to be similar to that of a still solution. Previously reported results show that 4RepCT silk coatings obtained during QCM-D measurements under the same conditions as in our studies result in randomly oriented rather than aligned nanofibrils, confirmed by AFM 33. A buffer baseline was established for at least 30 min.…”

supporting

confidence: 88%

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Interfacial Behavior of Recombinant Spider Silk Protein Parts Reveals Cues on the Silk Assembly Mechanism

et al. 2018

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The mechanism of silk assembly, and thus the cues for the extraordinary properties of silk, can be explored by studying the simplest protein parts needed for the formation of silk-like materials. The recombinant spider silk protein 4RepCT, consisting of four repeats of polyalanine and glycine-rich segments (4Rep) and a globular C-terminal domain (CT), has previously been shown to assemble into silk-like fibers at the liquid-air interface. Herein, we study the interfacial behavior of the two parts of 4RepCT, revealing new details on how each protein part is crucial for the silk assembly. Interfacial rheology and quartz crystal microbalance with dissipation show that 4Rep interacts readily at the interfaces. However, organized nanofibrillar structures are formed only when 4Rep is fused to CT. A strong interplay between the parts to direct the assembly is demonstrated. The presence of either a liquid-air or a liquid-solid interface had a surprisingly similar influence on the assembly.

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supporting

confidence: 88%

“…QSX-301 Au sensors (Biolin Scientific) for quartz crystal microbalance with dissipation (QCM-D) were cleaned and rendered hydrophobic using 2 mM 1-undecanethiol as previously described. 33…”

Section: Solid Surfacesmentioning

confidence: 99%

Interfacial Behavior of Recombinant Spider Silk Protein Parts Reveals Cues on the Silk Assembly Mechanism

et al. 2018

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“…Almost all studies reported that silk coatings increased smoothness and rendered the surfaces more hydrophilic . Although hydrophilic, it was found that 4RepCT coating formed a nanofibrillar structure . However, perhaps the most exciting result of this study was the nearly undetectable immune response as well as the slow biodegradation in vivo.…”

Section: Implant Coatingsmentioning

confidence: 63%

“…The silk coating assembled into fibrils on the surface, and these nanofibrous coatings, especially FN‐4RepCT, improved viability, spreading, and proliferation of HDMEC and human dermal fibroblasts. A decrease in S. aureus density was also observed on both 4RepCT and Mag‐4RepCT …”

Section: Implant Coatingsmentioning

confidence: 73%

Biomedical Applications of Recombinant Silk‐Based Materials

Aigner

DeSimone

Scheibel³

2018

Advanced Materials

251

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Silk is mostly known as a luxurious textile, which originates from silkworms first cultivated in China. A deeper look into the variety of silk reveals that it can be used for much more, in nature and by humanity. For medical purposes, natural silks were recognized early as a potential biomaterial for surgical threads or wound dressings; however, as biomedical engineering advances, the demand for high-performance, naturally derived biomaterials becomes more pressing and stringent. A common problem of natural materials is their large batch-to-batch variation, the quantity available, their potentially high immunogenicity, and their fast biodegradation. Some of these common problems also apply to silk; therefore, recombinant approaches for producing silk proteins have been developed. There are several research groups which study and utilize various recombinantly produced silk proteins, and many of these have also investigated their products for biomedical applications. This review gives a critical overview over of the results for applications of recombinant silk proteins in biomedical engineering.

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“…It has previously been reported that silk proteins adsorb at a constantly increasing rate on a surface and, through protein− protein interactions, form patterns similar to the ECM within connective tissues. 11 Thereby, added cells can sense a microenvironment similar to the native and develop essential cell-ECM attachment points through cell receptors. 10 Many of these cell receptors are transmembrane proteins (e.g., integrins) that bind to ECM proteins (e.g., fibronectin and laminin) and key mediators in cell adherence, migration, and proliferation.…”

Section: Discussionmentioning

confidence: 99%

Surface Functionalization of PTFE Membranes Intended for Guided Bone Regeneration Using Recombinant Spider Silk

Tasiopoulos

Petronis

Sahlin³

et al. 2019

ACS Appl. Bio Mater.

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Alveolar bone loss is usually treated with guided bone regeneration, a dental procedure which utilizes a tissue-separation membrane. The barrier membrane prevents pathogens and epithelial cells to invade the bone augmentation site, thereby permitting osteoblasts to deposit minerals and build up bone. This study aims at adding bioactive properties to otherwise inert PTFE membranes in order to enhance cell adherence and promote proliferation. A prewetting by ethanol and stepwise hydration protocol was herein employed to overcome high surface tension of PTFE membranes and allow for a recombinant spider silk protein, functionalized with a cellbinding motif from fibronectin (FN-silk), to self-assemble into a nanofibrillar coating. HaCaT and U-2 OS cells were seeded onto soft and hard tissue sides, respectively, of membranes coated with FN-silk. The cells could firmly adhere as early as 1 h post seeding, as well as markedly grow in numbers when kept in culture for 7 days. Fluorescence and scanning electron microscopy images revealed that adherent cells could form a confluent monolayer and develop essential cell−cell contacts during 1 week of culture. Hence, functionalized PTFE membranes have a potential of better integration at the implantation site, with reduced risk of membrane displacement as well as exposure to oral pathogens.

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Self-Assembly of Recombinant Silk as a Strategy for Chemical-Free Formation of Bioactive Coatings: A Real-Time Study

Cited by 52 publications

References 40 publications

Interfacial Behavior of Recombinant Spider Silk Protein Parts Reveals Cues on the Silk Assembly Mechanism

Interfacial Behavior of Recombinant Spider Silk Protein Parts Reveals Cues on the Silk Assembly Mechanism

Biomedical Applications of Recombinant Silk‐Based Materials

Surface Functionalization of PTFE Membranes Intended for Guided Bone Regeneration Using Recombinant Spider Silk

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