Photocrosslinkable natural polymers in tissue engineering

Moon, Seo Hyung; Hwang, Hye Jin; Jeon, Hye Ryeong; Park, Sol Ji; Bae, In Sun; Yang, Yun Jung

doi:10.3389/fbioe.2023.1127757

Cited by 20 publications

(12 citation statements)

References 190 publications

(263 reference statements)

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“…A more rapid photo-crosslinking process minimizes the opportunity for monomers or oligomers to diffuse from the intended area, thereby ensuring more precise control over polymerization sites. This precision facilitates the creation of finer features and more defined boundaries, leading to an enhancement in resolution [64,65].…”

Section: Resultsmentioning

confidence: 99%

3D bioprinting of Liver Microenvironment Model Using Photocrosslinkable Decellularized Extracellular Matrix based Hydrogel

Tabatabaei Rezaei,

Kumar,

Liu

et al. 2024

Preprint

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The liver, as one of the vital organs in the body, plays a crucial role in various bodily functions. Numerous factors can cause liver damage, that the sole remedy for severe liver conditions is the transplantation of healthy liver tissue. In response to the transplantation challenges, innovative approaches involving hydrogel-based technologies have emerged, leading to the creation of highly functionalized tissues. The development of three-dimensional printing and patterning of cell-laden biomaterial matrices offers promising advances for creating tissue-specific structures in tissue engineering and bioprinting. However, the matrix materials currently employed in bioprinting liver microtissue often fail to capture the complexity of the natural extracellular matrix (ECM), hindering their ability to restore innate cellular shapes and functions. Liver ECM-based hydrogels are increasingly recognized for their potential as biomimetic 3D cell culture systems that facilitate the exploration of liver disease, metabolism, and toxicity mechanisms. Yet, the conventional production of these hydrogels relies on slow thermal gelation processes, which restrict the manipulation of their mechanical characteristics. In this research, we introduce a novel approach with a functionalized photocrosslinkable liver decellularized extracellular matrix (dECM). By combining liver dECM methacrylate (LdECMMA) with gelatin methacrylate (GelMA), we achieved accelerated crosslinking under visible light irradiation and the ability to tune the mechanical, rheological, and physiological properties of the material. We encapsulated human hepatocellular carcinoma cells within an optimal concentration of the GelMA-LdECMMA hybrid hydrogel and examined cell proliferation and function over an extended period. The findings revealed that the GelMA-LdECMMA hybrid hydrogel enhances liver cell proliferation and function, holding significant promise for applications in drug screening and liver cancer metastasis research.

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Section: Resultsmentioning

confidence: 99%

3D bioprinting of Liver Microenvironment Model Using Photocrosslinkable Decellularized Extracellular Matrix based Hydrogel

Tabatabaei Rezaei,

Kumar,

Liu

et al. 2024

Preprint

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“…Introduction of methacryloyl groups is a popular route to expand the cross-linking repertoire of biopolymers while retaining biocompatibility. Additionally, this modification facilitates photopolymerization, which allows light-based 3D printing [ 93 ]. Methacrylation has been achieved for α-zein by reaction with glycidyl methacrylate (GMA) which introduced an average of 3.5 methacryloyl groups per zein monomer in mixtures of 70% ethanol and 30% water [ 94 ].…”

Section: Resultsmentioning

confidence: 99%

Conformations of a highly expressed Z19 α-zein studied with AlphaFold2 and MD simulations

Christensen

2024

PLoS ONE

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α-zeins are amphiphilic maize seed storage proteins with material properties suitable for a multitude of applications e.g., in renewable plastics, foods, therapeutics and additive manufacturing (3D-printing). To exploit their full potential, molecular-level insights are essential. The difficulties in experimental atomic-resolution characterization of α-zeins have resulted in a diversity of published molecular models. However, deep-learning α-zein models are largely unexplored. Therefore, this work studies an AlphaFold2 (AF2) model of a highly expressed α-zein using molecular dynamics (MD) simulations. The sequence of the α-zein cZ19C2 gave a loosely packed AF2 model with 7 α-helical segments connected by turns/loops. Compact tertiary structure was limited to a C-terminal bundle of three α-helices, each showing notable agreement with a published consensus sequence. Aiming to chart possible α-zein conformations in practically relevant solvents, rather than the native solid-state, the AF2 model was subjected to MD simulations in water/ethanol mixtures with varying ethanol concentrations. Despite giving structurally diverse endpoints, the simulations showed several patterns: In water and low ethanol concentrations, the model rapidly formed compact globular structures, largely preserving the C-terminal bundle. At ≥ 50 mol% ethanol, extended conformations prevailed, consistent with previous SAXS studies. Tertiary structure was partially stabilized in water and low ethanol concentrations, but was disrupted in ≥ 50 mol% ethanol. Aggregated results indicated minor increases in helicity with ethanol concentration. β-sheet content was consistently low (∼1%) across all conditions. Beyond structural dynamics, the rapid formation of branched α-zein aggregates in aqueous environments was highlighted. Furthermore, aqueous simulations revealed favorable interactions between the protein and the crosslinking agent glycidyl methacrylate (GMA). The proximity of GMA epoxide carbons and side chain hydroxyl oxygens simultaneously suggested accessible reactive sites in compact α-zein conformations and pre-reaction geometries for methacrylation. The findings may assist in expanding the applications of these technologically significant proteins, e.g., by guiding chemical modifications.

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“…Introduction of methacryloyl groups is a popular route to expand the cross-linking repertoire of biopolymers while retaining biocompatibility. Additionally, this modification facilitates photopolymerization, which allows light-based 3D printing [90]. Methacrylation has been achieved for α-zein by reaction with glycidyl methacrylate (GMA) which introduced an average of 3.5 methacryloyl groups per zein monomer in mixtures of 70% ethanol and 30% water [91].…”

Section: Resultsmentioning

confidence: 99%

Conformations of a highly expressed Z19 α-zein studied with AlphaFold2 and MD simulations

Christensen

2023

Preprint

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Alpha-zeins are amphiphilic maize proteins with interesting material properties suitable for numerous applications, e.g., in renewable plastics, foods, therapeutics and additive manufacturing (3D-printing). To exploit their full potential, molecular-level insights are essential. Since alpha-zeins have resisted experimental atomic-resolution characterization, molecular models have been central to elucidating their structure, but deep-learning alpha-zein models are largely unexplored. Therefore, this work studies an AlphaFold2 model of a highly expressed zein with molecular dynamics (MD) simulations. The mature protein sequence of the alpha-zein cZ19C2 gave a loosely packed model with 7 alpha-helical segments connected by turns/loops and 68% total alpha-helicity. Compact tertiary structure was limited to a C-terminal bundle of three alpha-helices, each aligning well with the published repeat sequence NPAAYLQQQQLLPFNQLA(V/A)(L/A). The model was subjected to MD simulations in water containing 0, 3, 23, 50, and 100 mol% ethanol for 400 ns, with extension of selected simulations to the us timescale. Although the simulations gave structurally diverse endpoints, several patterns were observed: In water and less than or equal to 2 mol% ethanol, the model rapidly formed compact globular structures, largely preserving the C-terminal bundle. At greater than or equal to 50 mol% ethanol, extended conformations prevailed, consistent with previous SAXS studies. Tertiary structure was partially stabilized in water and in 2 mol% and 23 mol% ethanol, but was disrupted in greater than or equal to 50 mol% ethanol. Averaged results indicated slightly increased helicity with ethanol concentration. Multiple copies of a globular model conformation rapidly formed branched aggregates in aqueous all-atom and coarse-grained MD simulations. Beta-sheet content was typically <1% across all simulations. In aqueous simulations with glycidyl methacrylate (GMA), 55% of GMA was found within 4 Angstrom of the model. Pre-reaction complexes for methacrylation were indicated by GMA epoxide carbons within 2.9 - 3.3 Angstrom of side chain hydroxyl oxygens, suggesting accessibility of reactive sites in compact alpha-zein conformations.

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Photocrosslinkable natural polymers in tissue engineering

Cited by 20 publications

References 190 publications

3D bioprinting of Liver Microenvironment Model Using Photocrosslinkable Decellularized Extracellular Matrix based Hydrogel

3D bioprinting of Liver Microenvironment Model Using Photocrosslinkable Decellularized Extracellular Matrix based Hydrogel

Conformations of a highly expressed Z19 α-zein studied with AlphaFold2 and MD simulations

Conformations of a highly expressed Z19 α-zein studied with AlphaFold2 and MD simulations

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