Qualitative comparison between different biopolymers for usage in two-photon polymerization towards liver regeneration

Ežerskytė, Eglė; Vengris, Mikas; Gineitis, Karolis; Merkininkaitė, Greta; Leber, Bettina; Vargalis, Rokas; STIEGLER, PHILIPP; Schemmer, Peter; Šakirzanovas, Simas; Kielaitė-Gulla, Aiste; Strupas, Kęstutis; Jonušauskas, Linas

doi:10.1364/ome.459057

Cited by 7 publications

(4 citation statements)

References 54 publications

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“…Previous reports indicated LAP to be sensitive to two-photon-based activation; [52][53][54] thus, we hypothesized that our DN system could enable the creation of complex 3D thiol-ene structures supported by the surrounding SPAAC matrix. Since LAP is only weakly two-photon active, we utilized rhodamine as a photosensitizer in conjunction with the photoinitiator to photopolymerize the second network in a set of intertwining rings via laser-scanning multiphoton lithography (Figure 4e, Supplementary Figure S3).…”

Section: Resultsmentioning

confidence: 99%

Stepwise Stiffening/Softening of and Cell Recovery from Reversibly Formulated Hydrogel Double Networks

Kopyeva,

Goldner,

Hoye

et al. 2024

Preprint

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Biomechanical contributions of the ECM underpin cell growth and proliferation, differentiation, signal transduction, and other fate decisions. As such, biomaterials whose mechanics can be spatiotemporally altered – particularly in a reversible manner – are extremely valuable for studying these mechanobiological phenomena. Herein, we introduce a poly(ethylene glycol) (PEG)-based hydrogel model consisting of two interpenetrating step-growth networks that are independently formed via largely orthogonal bioorthogonal chemistries and sequentially degraded with distinct bacterial transpeptidases, affording reversibly tunable stiffness ranges that span healthy and diseased soft tissues (e.g., 500 Pa – 6 kPa) alongside terminal cell recovery for pooled and/or single-cell analysis in a near “biologically invisible” manner. Spatiotemporal control of gelation within the primary supporting network was achieved via mask-based and two-photon lithography; these stiffened patterned regions could be subsequently returned to the original soft state following sortase-based secondary network degradation. Using this approach, we investigated the effects of 4D-triggered network mechanical changes on human mesenchymal stem cell (hMSC) morphology and Hippo signaling, as well as Caco-2 colorectal cancer cell mechanomemory at the global transcriptome level via RNAseq. We expect this platform to be of broad utility for studying and directing mechanobiological phenomena, patterned cell fate, as well as disease resolution in softer matrices.TOC DescriptionBiomaterials that can dynamically change stiffnesses are essential in further understanding the role of extracellular matrix mechanics. Using independently formulated and subsequently degradable interpenetrating hydrogel networks, we reversibly and spatiotemporally trigger stiffening/softening of cell-laden matrices. Terminal cell recovery for pooled and/or single-cell analysis is permitted in a near “biologically invisible” manner.

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Section: Resultsmentioning

confidence: 99%

Stepwise Stiffening/Softening of and Cell Recovery from Reversibly Formulated Hydrogel Double Networks

Kopyeva,

Goldner,

Hoye

et al. 2024

Preprint

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“…A series of BSA- GMA powders were synthesized by the methacrylation reaction, followed by dialysis and freeze-drying. LAP has good water solubility, good biocompatibility, and a two- photon absorption (TPA) of 0.16 GM at 800 nm, which is promising for TPP initiator [ 54 , 55 ] . Therefore, we chose LAP as the type I initiator for this study.…”

Section: Resultsmentioning

confidence: 99%

Synthesis of biocompatible BSA-GMA and two-photon polymerization of 3D hydrogels with free radical type I photoinitiator

Li¹,

Liu²,

Guo³

et al. 2023

IJB

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Although the development of three-dimensional (3D) printing technology is growing rapidly in the biomedical field, it remains a challenge to achieve arbitrary 3D structures with high resolution and high efficiency. Protein hydrogels fabricated by two-photon polymerization (TPP) have excellent mechanical properties, high precision, and 3D architecture. However, a large number of the amino acid group in bovine serum albumin (BSA) would be consumed when the protein-based hydrogels use dyes of free radical type II photoinitiators. In this study, we use glycidyl methacrylate (GMA) to modify BSA molecules to obtain a series of BSA-GMA materials, allowing the protein material to be two-photon polymerized with a water-soluble free radical type I photoinitiator. The precisely controllable 3D structure of the BSA-GMA hydrogel was fabricated by adjusting the concentration of the precursor solution, the degree of methacrylation, and the processing parameters of the TPP technique. Importantly, BSA-GMA materials are free of acidic hazardous substances. Meanwhile, the water-soluble initiator lithium phenyl (2,4,6-trimethylbenzoyl) phosphite (LAP) allows TPP on the vinyl group of the GMA chain and thus without consuming its amino acid group. The as-prepared BSA-GMA hydrogel structure exhibits excellent autofluorescence imaging, pH responsiveness, and biocompatibility, which would provide new avenues for potential applications in tissue engineering and biomedical fields to meet specific biological requirements.

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“…However, it was shown in numerous works that a wide array of materials are compatible with 2PP technology. 15,16 The sample is also illuminated using LED, which acts as a light source for a built-in imaging system consisting of a focusing lens and CMOS camera. For intensity distribution experiments CCD camera was placed into an optical chain into a focal plane of the first 4F lens.…”

Section: Methodsmentioning

confidence: 99%

Beam-shaping enhanced two-photon polymerization: towards 3D medical printing

Jonušauskas,

Vitkūnaitė,

Andrijec

et al. 2024

Laser 3D Manufacturing XI

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Two-photon polymerization (2PP) based 3D printing is a well-established technique. However, for the vast majority of its existence, 2PP was realized by applying general-purpose setups that were not tailored to any specific field. This resulted in limitations regarding how much 2PP can proliferate in any one particular area. Therefore, in this work, we will explore what can be achieved if a 2PP setup is built from the ground up for usage in one specific field -biomedical 3D printing. To achieve it a special femtosecond (fs) laser-based setup is assembled with integrated dynamic beam-shaping. Removal of zero order maximum in such an optical setup is demonstrated, and main related peculiarities are discussed. Then the beam-shaping is used to elongate voxels allowing to improve manufacturing throughput by more than two orders of magnitude. We then used the setup and developed voxel elongation methodology to produce biology-oriented structures, such as stents, grafts, and organoid scaffolds. We show excellent biocompatibility and cell growth on the later structures. Overall, the presented results show how focusing on 2PP system design to accommodate one particular field, in this case, biomedicine, helps to exploit the system's capabilities beyond what general purpose 2PP setup could achieve.

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Qualitative comparison between different biopolymers for usage in two-photon polymerization towards liver regeneration

Cited by 7 publications

References 54 publications

Stepwise Stiffening/Softening of and Cell Recovery from Reversibly Formulated Hydrogel Double Networks

Stepwise Stiffening/Softening of and Cell Recovery from Reversibly Formulated Hydrogel Double Networks

Synthesis of biocompatible BSA-GMA and two-photon polymerization of 3D hydrogels with free radical type I photoinitiator

Beam-shaping enhanced two-photon polymerization: towards 3D medical printing

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