Nanofibrillated cellulose/gellan gum hydrogel-based bioinks for 3D bioprinting of skin cells

Lameirinhas, Nicole S; Teixeira, M.C.; Carvalho, João Paulo Felicori; Valente, Bruno F. A.; Pinto, Ricardo J.B.; Oliveira, Helena; Luís, Jorge; Pires, Liliana; Oliveira, José M.; Vilela, Carla; Freire, Carmen S. R.

doi:10.1016/j.ijbiomac.2022.12.227

Cited by 23 publications

(13 citation statements)

References 63 publications

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“…Specifically, ALG:CEp_10% and ALG:CEpCUR_10% maintain good cell viabilities 1 day after the bioprinting procedure, with values of 76.8 ± 4.3% and 77.3 ± 1.9%, respectively. These values are very similar to the results obtained for the ALG hydrogel (76.6 ± 3.1%), proving that the presence of CEp and CEpCUR in the matrix does not , 44 about alginate hydrogels with lysozyme nanofibrils, that showed viabilities of nearly 88% after 7 days; and the study by Lameirinhas et al 67 regarding a bioink composed of gellan gum and cellulose nanofibers, with HaCaT cell viabilities of 90 ± 3%. 67 These results confirm that the composite alginate hydrogelbased bioinks with cellulose ester-based particles are adequate for 3D extrusion bioprinting of living constructs with HaCaT cells, maintaining a high cell viability up to 7 days after the 3D bioprinting procedure.…”

Section: D Printing Of Drug-releasing Structuressupporting

confidence: 84%

“…Other works concerning the 3D bioprinting of alginate-based hydrogels describe similar results, including the work of Lan et al that used TEMPO-oxidized cellulose/alginate hydrogels for the bioprinting of human meniscus fibrochondrocytes, and the study of Huang and colleagues where fibroblasts showed cell viabilities above 85% 2 days after bioprinting in a gelatin/sodium alginate/carboxymethyl chitosan bioink. Regarding the 3D bioprinting of HaCaT cells, our team has shown comparable outcomes using other bioink formulations, like the work of Teixeira et al, about alginate hydrogels with lysozyme nanofibrils, that showed viabilities of nearly 88% after 7 days; and the study by Lameirinhas et al regarding a bioink composed of gellan gum and cellulose nanofibers, with HaCaT cell viabilities of 90 ± 3% …”

Section: Resultsmentioning

confidence: 99%

“…These values are very similar to the results obtained for the ALG hydrogel (76.6 ± 3.1%), proving that the presence of CEp and CEpCUR in the matrix does not , 44 about alginate hydrogels with lysozyme nanofibrils, that showed viabilities of nearly 88% after 7 days; and the study by Lameirinhas et al 67 regarding a bioink composed of gellan gum and cellulose nanofibers, with HaCaT cell viabilities of 90 ± 3%. 67 These results confirm that the composite alginate hydrogelbased bioinks with cellulose ester-based particles are adequate for 3D extrusion bioprinting of living constructs with HaCaT cells, maintaining a high cell viability up to 7 days after the 3D bioprinting procedure. Moreover, the prior inclusion of a molecule of interest (i.e., a model drug) in the cellulose particles enables the creation of living constructs with drugreleasing ability, which constitutes an innovative approach for biomedical applications such as wound healing.…”

Section: D Printing Of Drug-releasing Structuresmentioning

confidence: 99%

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Hydrogel Bioinks of Alginate and Curcumin-Loaded Cellulose Ester-Based Particles for the Biofabrication of Drug-Releasing Living Tissue Analogs

Carvalho

Teixeira

Lameirinhas

et al. 2023

ACS Appl. Mater. Interfaces

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3D bioprinting is a versatile technique that allows the fabrication of living tissue analogs through the layer-by-layer deposition of cell-laden biomaterials, viz. bioinks. In this work, composite alginate hydrogel-based bioinks reinforced with curcumin-loaded particles of cellulose esters (CEpCUR) and laden with human keratinocytes (HaCaT) are developed. The addition of the CEpCUR particles, with sizes of 740 ± 147 nm, improves the rheological properties of the inks, increasing their shear stress and viscosity, while preserving the recovery rate and the mechanical and viscoelastic properties of the resulting fully cross-linked hydrogels. Moreover, the presence of these particles reduces the degradation rate of the hydrogels from 26.3 ± 0.8% (ALG) to 18.7 ± 1.3% (ALG:CEpCUR_10%) after 3 days in the culture medium. The 3D structures printed with the ALG:CEpCUR inks reveal increased printing definition and the ability to release curcumin (with nearly 70% of cumulative release after 24 h in PBS). After being laden with HaCaT cells (1.2 × 10 6 cells mL −1 ), the ALG:CEpCUR bioinks can be successfully 3D bioprinted, and the obtained living constructs show good dimensional stability and high cell viabilities at 7 days post-bioprinting (nearly 90%), confirming their great potential for application in fields like wound healing.

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Section: D Printing Of Drug-releasing Structuressupporting

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Section: D Printing Of Drug-releasing Structuresmentioning

confidence: 99%

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Hydrogel Bioinks of Alginate and Curcumin-Loaded Cellulose Ester-Based Particles for the Biofabrication of Drug-Releasing Living Tissue Analogs

Carvalho

Teixeira

Lameirinhas

et al. 2023

ACS Appl. Mater. Interfaces

Self Cite

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“…Few studies reported the keratinocyte-laden bioink printing for the epidermal layer biofabrication [ 48 , 49 ]. Lameirinhas et al investigated the viability of 3D bioprinted construct using nanofibrillated cellulose/gellan gum hydrogel-based bioinks loaded with HaCaT cell [ 49 ].…”

Section: Discussionmentioning

confidence: 99%

Marine Collagen-Based Bioink for 3D Bioprinting of a Bilayered Skin Model

et al. 2023

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Marine organisms (i.e., fish, jellyfish, sponges or seaweeds) represent an abundant and eco-friendly source of collagen. Marine collagen, compared to mammalian collagen, can be easily extracted, is water-soluble, avoids transmissible diseases and owns anti-microbial activities. Recent studies have reported marine collagen as a suitable biomaterial for skin tissue regeneration. The aim of this work was to investigate, for the first time, marine collagen from basa fish skin for the development of a bioink for extrusion 3D bioprinting of a bilayered skin model. The bioinks were obtained by mixing semi-crosslinked alginate with 10 and 20 mg/mL of collagen. The bioinks were characterised by evaluating the printability in terms of homogeneity, spreading ratio, shape fidelity and rheological properties. Morphology, degradation rate, swelling properties and antibacterial activity were also evaluated. The alginate-based bioink containing 20 mg/mL of marine collagen was selected for 3D bioprinting of skin-like constructs with human fibroblasts and keratinocytes. The bioprinted constructs showed a homogeneous distribution of viable and proliferating cells at days 1, 7 and 14 of culture evaluated by qualitative (live/dead) and qualitative (XTT) assays, and histological (H&E) and gene expression analysis. In conclusion, marine collagen can be successfully used to formulate a bioink for 3D bioprinting. In particular, the obtained bioink can be printed in 3D structures and is able to support fibroblasts and keratinocytes viability and proliferation.

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“…3D printing, also known as additive manufacturing, is a digital manufacturing technology that fabricates components layer by layer according to a specific path [28,29], which has been widely used to construct human tissues/organs with highly bionic structures and components [8]. The core idea of 3D printing is to realize the controllable spatial distribution of biological materials, cells and other active substances [30][31][32], which has the advantages of strong controllability, short production cycle, and individual customization [33]. In one study, silk fibroin-gelatin bioink was extruded to print unique RRs characteristics by adjusting printing parameters.…”

Section: Introductionmentioning

confidence: 99%

Biofabrication of biomimetic undulating microtopography at the dermal-epidermal junction and its effects on the growth and differentiation of epidermal cells

Gao,

Lu,

Liu

et al. 2024

Biofabrication

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The undulating microtopography located at the junction of the dermis and epidermis of the native skin is called rete ridges (RRs), which plays an important role in enhancing keratinocyte function, improving skin structure and stability, and providing three-dimensional (3D) microenvironment for skin cells. Despite some progress in recent years, most currently designed and manufactured tissue-engineered skin models still cannot replicate the RRs, resulting in a lack of biological signals in the manufactured skin models. In this study, a composite manufacturing method including electrospinning, 3D printing, and functional coating was developed to produce the epidermal models with RRs. Polycaprolactone (PCL) nanofibers were firstly electrospun to mimic the extracellular matrix environment and be responsible for cell attachment. PCL microfibers were then printed onto top of the PCL nanofibers layer by 3D printing to quickly prepare undulating microtopography and finally the entire structures were dip-coated with gelatin hydrogel to form a functional coating layer. The morphology, chemical composition, and structural properties of the fabricated models were studied. The results proved that the multi-process composite fabricated models were suitable for skin tissue engineering. Live and dead staining, cell counting kit-8 (CCK-8) as well as histology (haematoxylin and eosin (HE) methodology) and immunofluorescence (primary and secondary antibodies combination assay) were used to investigate the viability, metabolic activity, and differentiation of skin cells for in vitro culturing. In vitro results showed that each model had high cell viability, good proliferation, and the expression of differentiation marker. It was worth noting that the sizes of the RRs affected the cell growth status of the epidermal models. In addition, the unique undulation characteristics of the epidermal-dermal junction can be reproduced in the developed epidermal models. Overall, these in vitro human epidermal models can provide valuable reference for skin transplantation, screening and safety evaluation of drugs and cosmetics.

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Nanofibrillated cellulose/gellan gum hydrogel-based bioinks for 3D bioprinting of skin cells

Cited by 23 publications

References 63 publications

Hydrogel Bioinks of Alginate and Curcumin-Loaded Cellulose Ester-Based Particles for the Biofabrication of Drug-Releasing Living Tissue Analogs

Hydrogel Bioinks of Alginate and Curcumin-Loaded Cellulose Ester-Based Particles for the Biofabrication of Drug-Releasing Living Tissue Analogs

Marine Collagen-Based Bioink for 3D Bioprinting of a Bilayered Skin Model

Biofabrication of biomimetic undulating microtopography at the dermal-epidermal junction and its effects on the growth and differentiation of epidermal cells

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