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47 pages, 30956 KiB

Open AccessArticle

Screening of Self-Assembling of Collagen IV Fragments into Stable Structures Potentially Useful in Regenerative Medicine

by Marcin Kolasa, Grzegorz Galita, Ireneusz Majsterek, Ewa Kucharska, Katarzyna Czerczak, Joanna Wasko, Angelika Becht, Justyna Fraczyk, Anna Gajda, Lukasz Pietrzak, Lukasz Szymanski, Agnieszka Krakowiak, Zbigniew Draczynski and Beata Kolesinska

Int. J. Mol. Sci. 2021, 22(24), 13584; https://doi.org/10.3390/ijms222413584 - 18 Dec 2021

Cited by 2 | Viewed by 2620

Abstract

The aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV [...] Read more.

The aim of the research was to check whether it is possible to use fragments of type IV collagen to obtain, as a result of self-assembling, stable spatial structures that could be used to prepare new materials useful in regenerative medicine. Collagen IV fragments were obtained by using DMT/NMM/TosO⁻ as a coupling reagent. The ability to self-organize and form stable spatial structures was tested by the CD method and microscopic techniques. Biological studies covered: resazurin assay (cytotoxicity assessment) on BJ, BJ-5TA and C2C12 cell lines; an alkaline version of the comet assay (genotoxicity), Biolegend Legendplex human inflammation panel 1 assay (SC cell lines, assessment of the inflammation activity) and MTT test to determine the cytotoxicity of the porous materials based on collagen IV fragments. It was found that out of the pool of 37 fragments (peptides 1–33 and 2.1–2.4) reconstructing the outer sphere of collagen IV, nine fragments (peptides: 2, 4, 5, 6, 14, 15, 25, 26 and 30), as a result of self-assembling, form structures mimicking the structure of the triple helix of native collagens. The stability of spatial structures formed as a result of self-organization at temperatures of 4 °C, 20 °C, and 40 °C was found. The application of the MST method allowed us to determine the K_d of binding of selected fragments of collagen IV to ITGα1β1. The stability of the spatial structures of selected peptides made it possible to obtain porous materials based on their equimolar mixture. The formation of the porous materials was found for cross-linked structures and the material stabilized only by weak interactions. All tested peptides are non-cytotoxic against all tested cell lines. Selected peptides also showed no genotoxicity and no induction of immune system responses. Research on the use of porous materials based on fragments of type IV collagen, able to form stable spatial structures as scaffolds useful in regenerative medicine, will be continued. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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16 pages, 2526 KiB

Open AccessArticle

Docosahexaenoic Acid Modulates Paracellular Absorption of Testosterone and Claudin-1 Expression in a Tissue-Engineered Skin Model

by Andréa Tremblay, Mélissa Simard, Sophie Morin and Roxane Pouliot

Int. J. Mol. Sci. 2021, 22(23), 13091; https://doi.org/10.3390/ijms222313091 - 03 Dec 2021

Cited by 1 | Viewed by 2026

Abstract

Healthy skin moLEdels produced by tissue-engineering often present a suboptimal skin barrier function as compared with normal human skin. Moreover, skin substitutes reconstructed according to the self-assembly method were found to be deficient in polyunsaturated fatty acids (PUFAs). Therefore, in this study, we [...] Read more.

Healthy skin moLEdels produced by tissue-engineering often present a suboptimal skin barrier function as compared with normal human skin. Moreover, skin substitutes reconstructed according to the self-assembly method were found to be deficient in polyunsaturated fatty acids (PUFAs). Therefore, in this study, we investigated the effects of a supplementation of the culture media with docosahexaenoic acid (DHA) on the barrier function of skin substitutes. To this end, 10 μM DHA-supplemented skin substitutes were produced (n = 3), analyzed, and compared with controls (substitutes without supplementation). A Franz cell diffusion system, followed by ultra-performance liquid chromatography, was used to perform a skin permeability to testosterone assay. We then used gas chromatography to quantify the PUFAs found in the epidermal phospholipid fraction of the skin substitutes, which showed successful DHA incorporation. The permeability to testosterone was decreased following DHA supplementation and the lipid profile was improved. Differences in the expression of the tight junction (TJ) proteins claudin-1, claudin-4, occludin, and TJ protein-1 were observed, principally a significant increase in claudin-1 expression, which was furthermore confirmed by Western blot analyses. In conclusion, these results confirm that the DHA supplementation of cell culture media modulates different aspects of skin barrier function in vitro and reflects the importance of n-3 PUFAs regarding the lipid metabolism in keratinocytes. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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12 pages, 1292 KiB

Open AccessArticle

Encapsulin Based Self-Assembling Iron-Containing Protein Nanoparticles for Stem Cells MRI Visualization

by Anna N. Gabashvili, Stepan S. Vodopyanov, Nelly S. Chmelyuk, Viktoria A. Sarkisova, Konstantin A. Fedotov, Maria V. Efremova and Maxim A. Abakumov

Int. J. Mol. Sci. 2021, 22(22), 12275; https://doi.org/10.3390/ijms222212275 - 12 Nov 2021

Cited by 8 | Viewed by 2531

Abstract

Over the past decade, cell therapy has found many applications in the treatment of different diseases. Some of the cells already used in clinical practice include stem cells and CAR-T cells. Compared with traditional drugs, living cells are much more complicated systems that [...] Read more.

Over the past decade, cell therapy has found many applications in the treatment of different diseases. Some of the cells already used in clinical practice include stem cells and CAR-T cells. Compared with traditional drugs, living cells are much more complicated systems that must be strictly controlled to avoid undesirable migration, differentiation, or proliferation. One of the approaches used to prevent such side effects involves monitoring cell distribution in the human body by any noninvasive technique, such as magnetic resonance imaging (MRI). Long-term tracking of stem cells with artificial magnetic labels, such as magnetic nanoparticles, is quite problematic because such labels can affect the metabolic process and cell viability. Additionally, the concentration of exogenous labels will decrease during cell division, leading to a corresponding decrease in signal intensity. In the current work, we present a new type of genetically encoded label based on encapsulin from Myxococcus xanthus bacteria, stably expressed in human mesenchymal stem cells (MSCs) and coexpressed with ferroxidase as a cargo protein for nanoparticles’ synthesis inside encapsulin shells. mZip14 protein was expressed for the enhancement of iron transport into the cell. Together, these three proteins led to the synthesis of iron-containing nanoparticles in mesenchymal stem cells—without affecting cell viability—and increased contrast properties of MSCs in MRI. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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15 pages, 3961 KiB

Open AccessArticle

Adhesive Antimicrobial Peptides Containing 3,4-Dihydroxy-L-Phenylalanine Residues for Direct One-Step Surface Coating

by Young Eun Hwang, Seonghun Im, Hyun Kim, Jung-Hoon Sohn, Byung-Kwan Cho, Ju Hyun Cho, Bong Hyun Sung and Sun Chang Kim

Int. J. Mol. Sci. 2021, 22(21), 11915; https://doi.org/10.3390/ijms222111915 - 03 Nov 2021

Cited by 8 | Viewed by 2304

Abstract

Bacterial colonization and transmission via surfaces increase the risk of infection. In this study, we design and employ novel adhesive antimicrobial peptides to prevent bacterial contamination of surfaces. Repeats of 3,4-dihydroxy-L-phenylalanine (DOPA) were added to the C-terminus of NKC, a potent synthetic antimicrobial [...] Read more.

Bacterial colonization and transmission via surfaces increase the risk of infection. In this study, we design and employ novel adhesive antimicrobial peptides to prevent bacterial contamination of surfaces. Repeats of 3,4-dihydroxy-L-phenylalanine (DOPA) were added to the C-terminus of NKC, a potent synthetic antimicrobial peptide, and the adhesiveness and antibacterial properties of the resulting peptides are evaluated. The peptide is successfully immobilized on polystyrene, titanium, and polydimethylsiloxane surfaces within 10 min in a one-step coating process with no prior surface functionalization. The antibacterial effectiveness of the NKC-DOPA₅-coated polystyrene, titanium, and polydimethylsiloxane surfaces is confirmed by complete inhibition of the growth of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus within 2 h. The stability of the peptide coated on the substrate surface is maintained for 84 days, as confirmed by its bactericidal activity. Additionally, the NKC-DOPA₅-coated polystyrene, titanium, and polydimethylsiloxane surfaces show no cytotoxicity toward the human keratinocyte cell line HaCaT. The antimicrobial properties of the peptide-coated surfaces are confirmed in a subcutaneous implantation animal model. The adhesive antimicrobial peptide developed in this study exhibits potential as an antimicrobial surface-coating agent for efficiently killing a broad spectrum of bacteria on contact. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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12 pages, 2980 KiB

Open AccessArticle

Design of PG-Surfactants Bearing Polyacrylamide Polymer Chain to Solubilize Membrane Proteins in a Surfactant-Free Buffer

by Taro Shimamoto, Tatsuki Nakakubo, Tomoyasu Noji, Shuhei Koeda, Keisuke Kawakami, Nobuo Kamiya and Toshihisa Mizuno

Int. J. Mol. Sci. 2021, 22(4), 1524; https://doi.org/10.3390/ijms22041524 - 03 Feb 2021

Cited by 1 | Viewed by 1930

Abstract

The development of techniques capable of using membrane proteins in a surfactant-free aqueous buffer is an attractive research area, and it should be elucidated for various membrane protein studies. To this end, we examined a method using new solubilization surfactants that do not [...] Read more.

The development of techniques capable of using membrane proteins in a surfactant-free aqueous buffer is an attractive research area, and it should be elucidated for various membrane protein studies. To this end, we examined a method using new solubilization surfactants that do not detach from membrane protein surfaces once bound. The designed solubilization surfactants, DKDKC₁₂K-PA_n (n = 5, 7, and 18), consist of two parts: one is the lipopeptide-based solubilization surfactant part, DKDKC₁₂K, fand the other is the covalently connected linear polyacrylamide (PA) chain with different M_w values of 5, 7, or 18 kDa. Intermolecular interactions between the PA chains in DKDKC₁₂K-PA_n concentrated on the surfaces of membrane proteins via amphiphilic binding of the DKDKC₁₂K part to the integral membrane domain was observed. Therefore, DKDKC₁₂K-PA_n (n = 5, 7, and 18) could maintain a bound state even after removal of the unbound by ultrafiltration or gel-filtration chromatography. We used photosystem I (PSI) from Thermosynecoccus vulcanus as a representative to assess the impacts of new surfactants on the solubilized membrane protein structure and functions. Based on the maintenance of unique photophysical properties of PSI, we evaluated the ability of DKDKC₁₂K-PA_n (n = 5, 7, and 18) as a new solubilization surfactant. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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14 pages, 2524 KiB

Open AccessArticle

Surface Modification by Nano-Structures Reduces Viable Bacterial Biofilm in Aerobic and Anaerobic Environments

by Sarah Ya’ari, Michal Halperin-Sternfeld, Boris Rosin and Lihi Adler-Abramovich

Int. J. Mol. Sci. 2020, 21(19), 7370; https://doi.org/10.3390/ijms21197370 - 06 Oct 2020

Cited by 7 | Viewed by 4377

Abstract

Bacterial biofilm formation on wet surfaces represents a significant problem in medicine and environmental sciences. One of the strategies to prevent or eliminate surface adhesion of organisms is surface modification and coating. However, the current coating technologies possess several drawbacks, including limited durability, [...] Read more.

Bacterial biofilm formation on wet surfaces represents a significant problem in medicine and environmental sciences. One of the strategies to prevent or eliminate surface adhesion of organisms is surface modification and coating. However, the current coating technologies possess several drawbacks, including limited durability, low biocompatibility and high cost. Here, we present a simple antibacterial modification of titanium, mica and glass surfaces using self-assembling nano-structures. We have designed two different nano-structure coatings composed of fluorinated phenylalanine via the drop-cast coating technique. We investigated and characterized the modified surfaces by scanning electron microscopy, X-ray diffraction and wettability analyses. Exploiting the antimicrobial property of the nano-structures, we successfully hindered the viability of Streptococcus mutans and Enterococcus faecalis on the coated surfaces in both aerobic and anaerobic conditions. Notably, we found lower bacteria adherence to the coated surfaces and a reduction of 86–99% in the total metabolic activity of the bacteria. Our results emphasize the interplay between self-assembly and antimicrobial activity of small self-assembling molecules, thus highlighting a new approach of biofilm control for implementation in biomedicine and other fields. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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Review

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17 pages, 2747 KiB

Open AccessReview

Applications of DNA-Functionalized Proteins

by Zhaoqiu Gong, Yuanyuan Tang, Ningning Ma, Wenhong Cao, Yong Wang, Shuang Wang and Ye Tian

Int. J. Mol. Sci. 2021, 22(23), 12911; https://doi.org/10.3390/ijms222312911 - 29 Nov 2021

Cited by 1 | Viewed by 2172

Abstract

As an important component that constitutes all the cells and tissues of the human body, protein is involved in most of the biological processes. Inspired by natural protein systems, considerable efforts covering many discipline fields were made to design artificial protein assemblies and [...] Read more.

As an important component that constitutes all the cells and tissues of the human body, protein is involved in most of the biological processes. Inspired by natural protein systems, considerable efforts covering many discipline fields were made to design artificial protein assemblies and put them into application in recent decades. The rapid development of structural DNA nanotechnology offers significant means for protein assemblies and promotes their application. Owing to the programmability, addressability and accurate recognition ability of DNA, many protein assemblies with unprecedented structures and improved functions have been successfully fabricated, consequently creating many brand-new researching fields. In this review, we briefly introduced the DNA-based protein assemblies, and highlighted the limitations in application process and corresponding strategies in four aspects, including biological catalysis, protein detection, biomedicine treatment and other applications. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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23 pages, 1981 KiB

Open AccessReview

Self-Assembling Peptides: From Design to Biomedical Applications

by Sara La Manna, Concetta Di Natale, Valentina Onesto and Daniela Marasco

Int. J. Mol. Sci. 2021, 22(23), 12662; https://doi.org/10.3390/ijms222312662 - 23 Nov 2021

Cited by 41 | Viewed by 4289

Abstract

Self-assembling peptides could be considered a novel class of agents able to harvest an array of micro/nanostructures that are highly attractive in the biomedical field. By modifying their amino acid composition, it is possible to mime several biological functions; when assembled in micro/nanostructures, [...] Read more.

Self-assembling peptides could be considered a novel class of agents able to harvest an array of micro/nanostructures that are highly attractive in the biomedical field. By modifying their amino acid composition, it is possible to mime several biological functions; when assembled in micro/nanostructures, they can be used for a variety of purposes such as tissue regeneration and engineering or drug delivery to improve drug release and/or stability and to reduce side effects. Other significant advantages of self-assembled peptides involve their biocompatibility and their ability to efficiently target molecular recognition sites. Due to their intrinsic characteristics, self-assembled peptide micro/nanostructures are capable to load both hydrophobic and hydrophilic drugs, and they are suitable to achieve a triggered drug delivery at disease sites by inserting in their structure’s stimuli-responsive moieties. The focus of this review was to summarize the most recent and significant studies on self-assembled peptides with an emphasis on their application in the biomedical field. Full article

(This article belongs to the Special Issue Self-Assembling Peptide-Based Biomaterials: Smart Materials for Biomedical Applications)

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