Abstract:Polymer multicomponent coatings such as multilayers mimic extracellular matrix (ECM) that attracts significant attention for their use as functional supports for advanced cell culture and tissue engineering. Herein, biodegradation and molecular transport in hyaluronan/polylysine multilayers coated with gold nanoparticles was described. Nanoparticle 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 2 coating acts as semipermeable barrier that governs molecular transport into/from the multilayers and makes them biodegradation resistant. Model protein lysozyme (mimics of ECM soluble signals) diffuses in the multilayers as fast and slow diffusing populations existing in an equilibrium. Such composite system may have high potential to be exploited as degradationresistant drug delivery platforms suitable for cell-based applications.The extracellular matrix (ECM) provides not only a structural support for cellular growth, but also a biochemical milieu, which together define cell fate. 1 Artificial ECM is designed to mimic real ECM and to achieve cellular functions in laboratory designed materials similar as in nature.Artificial ECM consists of degradable supports made of natural or (semi)synthetic polymers which: i) recapitulate physical-chemical properties of real ECM, ii) host bioactive signaling molecules (e.g. hormones, cytokines, growth factors) to be presented to cells, and iii) possess tuned biodegradability. 2-3 The issues above are challenging for the fabrication of artificial ECM but are essential for successful applications in tissue engineering, diagnostics, and animal-free studies.Introduced in the early nineties, 4 layer-by-layer deposition proofed to be extremely useful in the field of bio-functional coatings. It is powerful technique for the fabrication of polymer-based multilayer films with dimensions from nano to micro, adjusted composition, and tunable cellular response. [5][6][7][8][9][10][11] Control over cellular functions such as adhesion, differentiation, and proliferation, 12-15 along with reservoir properties for a variety of biomolecules such as proteins, growth factors, genes, and drugs 7, 16-20 makes these films an attractive platform for cell-based applications.Despite the evident biological potential to employ the multilayers to recapitulate ECM, 3 the options to tune multilayer biodegradability are mostly limited to multilayer chemical 13,[21][22] that can significantly change multilayer properties. There is also a lack in understanding the mechanism of biomolecule-multilayer interaction that obviously defines biomolecule presentation/release from multilayers to cells and, as a result, cellular response. [23][24] In this work a new approach for the design of artificial ECM with controlled biodegradation based on the coating of multilayers with gold nanoparticles (GNPs) is present. The multilayers wer...
Comprehensive analysis of the multifractional molecular diffusion provides a deeper understanding of the diffusion phenomenon in the fields of material science, molecular and cell biology, advanced biomaterials, etc. Fluorescence recovery after photobleaching (FRAP) is commonly employed to probe the molecular diffusion. Despite FRAP being a very popular method, it is not easy to assess multifractional molecular diffusion due to limited possibilities of approaches for analysis. Here we present a novel simulation-optimization-based approach (S-approach) that significantly broadens possibilities of the analysis. In the S-approach, possible fluorescence recovery scenarios are primarily simulated and afterward compared with a real measurement while optimizing parameters of a model until a sufficient match is achieved. This makes it possible to reveal multifractional molecular diffusion. Fluorescent latex particles of different size and fluorescein isothiocyanate in an aqueous medium were utilized as test systems. Finally, the S-approach has been used to evaluate diffusion of cytochrome c loaded into multilayers made of hyaluronan and polylysine. Software for evaluation of multifractional molecular diffusion by S-approach has been developed aiming to offer maximal versatility and user-friendly way for analysis.
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