RGD density along with substrate stiffness regulate hPSC hepatocyte functionality through YAP signalling

Blackford, Samuel J.I.; Yu, Tracy T. L.; Norman, Michael D. A.; Syanda, Adam M.; Manolakakis, Michail; Lachowski, Dariusz; Guo, Yunzhe; Garitta, Elena; Riccio, Federica; Jowett, Geraldine M.; Ng, Soon Seng; Vérnia, Santiago; Hernández, Armando E. del Río; Gentleman, Eileen; Rashid, S. Tamir

doi:10.1016/j.biomaterials.2022.121982

Cited by 8 publications

(5 citation statements)

References 74 publications

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“…For the dECM/FGelMa scaffold, it was found that IL-10 expression was higher than that in the FGelMa hydrogel, indicating that a scaffold containing dECM can improve the anti-inflammatory properties of HCs. Primary cells loaded into a 3D hydrogel culture system showed enhanced YAP activity under the influence of tensile force and maintained differentiation [55]. Our data strongly demonstrate tensile force-associated and YAP-mediated regulation of HC behaviors including immune responses, morphology, proliferation, and chondrogenic characteristics.…”

Section: Effect Of Cyclic Tensile Stimulation On Gene Expressionsupporting

confidence: 64%

3D-biofabricated chondrocyte-laden decellularized extracellular matrix-contained gelatin methacrylate auxetic scaffolds under cyclic tensile stimulation for cartilage regeneration

Chen

Lin

et al. 2023

Biofabrication

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Three-dimensional hydrogel constructs can mimic features of the extracellular matrix and have tailorable physicochemical properties to support and maintain regeneration of articular cartilage. Various studies have shown that mechanical cues affect the cellular microenvironment and thereby influence cellular behavior. In this study, we fabricated an auxetic scaffold to investigate the effect of three-dimensional (3D) tensile stimulation on chondrocyte behavior. Different concentrations of decellularized extracellular matrix (dECM) were mixed with fish gelatin methacrylate (FGelMa) and employed for the preparation of dECM/FGelMa auxetic bio-scaffolds using 3D biofabrication technology. We show that when human chondrocytes (HC) were incorporated into these scaffolds, their proliferation and the expression of chondrogenesis-related markers increased with dECM content. The function of HC was influenced by cyclic tensile stimulation, as shown by increased production of the chondrogenesis-related markers, collagen II and glycosaminoglycans, with the involvement of the yes-associated protein 1 signaling pathway. The biofabricated auxetic scaffold represents an excellent platform for exploring interactions between cells and their mechanical microenvironment.

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Section: Effect Of Cyclic Tensile Stimulation On Gene Expressionsupporting

confidence: 64%

3D-biofabricated chondrocyte-laden decellularized extracellular matrix-contained gelatin methacrylate auxetic scaffolds under cyclic tensile stimulation for cartilage regeneration

Chen

Lin

et al. 2023

Biofabrication

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“…To create hydrogels containing the FRET sensor, we built from our existing design, which relies on a two-step conjugation reaction. [ 20 , 30 ] Heterobifunctional peptides that are either non-functional or contain an MMP-degradable or an RGD sequence are first conjugated to 4-arm PEG functionalized at each terminus with nitrophenyl carbonate through a nucleophilic substitution reaction with the primary amine on an N-terminal lysine ( Figure 3a ). The network is cross-linked through a Michael addition between a C-terminal cysteine on the peptide with PEG-4VS in a 1:1 stoichiometric ratio.…”

Section: Resultsmentioning

confidence: 99%

FRET Sensor‐Modified Synthetic Hydrogels for Real‐Time Monitoring of Cell‐Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

Yan,

Kavanagh,

da Silva Harrabi

et al. 2024

Adv Funct Materials

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Matrix remodeling plays central roles in a range of physiological and pathological processes and is driven predominantly by the activity of matrix metalloproteinases (MMPs), which degrade extracellular matrix (ECM) proteins. How MMPs regulate cell and tissue dynamics is not well understood as in vivo approaches are lacking and many in vitro strategies cannot provide high‐resolution, quantitative measures of enzyme activity in situ within tissue‐like 3D microenvironments. Here, a Förster resonance energy transfer (FRET) sensor of MMP activity is incorporated into fully synthetic hydrogels that mimic many properties of the native ECM. Fluorescence lifetime imaging is then used to provide a real‐time, fluorophore concentration‐independent quantification of MMP activity, establishing a highly accurate, readily adaptable platform for studying MMP dynamics in situ. MCF7 human breast cancer cells encapsulated within hydrogels are then used to detect MMP activity both locally, at the sub‐micron level, and within the bulk hydrogel. This versatile platform may find use in a range of biological studies to explore questions in the dynamics of cancer metastasis, development, and tissue repair by providing high‐resolution, quantitative, and in situ readouts of local MMP activity within native tissue‐like environments.

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“…Bioinks can be generated from natural components (e.g., decellularized liver ECM, collagen, gelatin, alginate) or synthetic polymers (e.g., polyethylene glycol, polycaprolactone, polyvinylpyrrolidone) ( Khoeini et al, 2021 ). Blackford et al (2023) reported that HLCs cultured on soft polyethylene glycol (2.5%) secreted more ALB than PHHs and metabolized more urea than HLCs cultured in alginate ( Blackford et al, 2023 ), underscoring the potential of synthetic polymers for bioprinting. Additionally, Abbasalizadeh et al (2023) achieved a high yield of hiPSC-derived hepatic organoids by combining PEG microencapsulation and microfluidic systems, paving the way for cell transplantation ( Abbasalizadeh et al, 2023 ).…”

Section: Hipsc-derived Hepatocytes For Bioprintingmentioning

confidence: 99%

iPSC-derived cells for whole liver bioengineering

Telles-Silva,

Pacheco,

Chianca

et al. 2024

Front. Bioeng. Biotechnol.

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Liver bioengineering stands as a prominent alternative to conventional hepatic transplantation. Through liver decellularization and/or bioprinting, researchers can generate acellular scaffolds to overcome immune rejection, genetic manipulation, and ethical concerns that often accompany traditional transplantation methods, in vivo regeneration, and xenotransplantation. Hepatic cell lines derived from induced pluripotent stem cells (iPSCs) can repopulate decellularized and bioprinted scaffolds, producing an increasingly functional organ potentially suitable for autologous use. In this mini-review, we overview recent advancements in vitro hepatocyte differentiation protocols, shedding light on their pivotal role in liver recellularization and bioprinting, thereby offering a novel source for hepatic transplantation. Finally, we identify future directions for liver bioengineering research that may allow the implementation of these systems for diverse applications, including drug screening and liver disease modeling.

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RGD density along with substrate stiffness regulate hPSC hepatocyte functionality through YAP signalling

Cited by 8 publications

References 74 publications

3D-biofabricated chondrocyte-laden decellularized extracellular matrix-contained gelatin methacrylate auxetic scaffolds under cyclic tensile stimulation for cartilage regeneration

3D-biofabricated chondrocyte-laden decellularized extracellular matrix-contained gelatin methacrylate auxetic scaffolds under cyclic tensile stimulation for cartilage regeneration

FRET Sensor‐Modified Synthetic Hydrogels for Real‐Time Monitoring of Cell‐Derived Matrix Metalloproteinase Activity using Fluorescence Lifetime Imaging

iPSC-derived cells for whole liver bioengineering

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