Fabrication of co-cultured tissue constructs using a dual cell seeding compatible cell culture insert with a clip-on scaffold for potential regenerative medicine and toxicological screening applications

Ameer, Jimna Mohamed; Venkatesan, Ramesh Babu; Damodaran, Vinod B.; Nishad, K. V.; Sabareeswaran, A; Asari, Anil Kumar Pallickaveedu Rajan; Kasoju, Naresh

doi:10.1016/j.jsamd.2020.04.004

Cited by 9 publications

(5 citation statements)

References 37 publications

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“…Immortalised cell lines, which commonly include human keratinocyte cells (HaCaT) and N/TERT cells, are financially viable and provide reasonably consistent results . However, HaCaT cells ,− fail to correctly express cornified envelope proteins in the differentiation cycle when compared to native human primary keratinocyte cells, to form the biomechanically critical stratum corneum. , Further comparisons of immortalized cells N/TERT1 and N/TERT2G to human primary keratinocytes show similar differentiation gene expression levels, and N/TERT immortalized cells have been presented as a more relevant substitute for primary keratinocytes than HaCaT cells . It is recognized that primary keratinocytes are the ideal choice of cell for accuracy of the skin model, as primary keratinocytes more accurately reflect in vivo cell behavior; however, primary keratinocytes require longer culture periods and undergo faster cell senescence when compared with immortalized cells …”

Section: Layer-by-layer Analysis Of Skin Tissue Modelsmentioning

confidence: 96%

“…191 Their ability of induced hair neogenesis is impaired in 2D systems, with increased essential gene expression observed in 3D culture. 69 Spheroid protocols have been followed to first form dermal papillae cell aggregates 107 that form hair follicles which show characteristic bulge-like regions 54 (Figure 2). Betriu et al used a RAD16-I (self-assembling peptide) scaffold aiming to preserve the expected phenotype of dermal papilla cultures using a biofabrication approach.…”

Section: Basement Membrane Zonementioning

confidence: 99%

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Layer-by-Layer Analysis of In Vitro Skin Models

Footner,

Firipis,

Liu

et al. 2023

ACS Biomater. Sci. Eng.

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In vitro human skin models are evolving into versatile platforms for the study of skin biology and disorders. These models have many potential applications in the fields of drug testing and safety assessment, as well as cosmetic and new treatment development. The development of in vitro skin models that accurately mimic native human skin can reduce reliance on animal models and also allow for more precise, clinically relevant testing. Recent advances in biofabrication techniques and biomaterials have led to the creation of increasingly complex, multilayered skin models that incorporate important functional components of skin, such as the skin barrier, mechanical properties, pigmentation, vasculature, hair follicles, glands, and subcutaneous layer. This improved ability to recapitulate the functional aspects of native skin enhances the ability to model the behavior and response of native human skin, as the complex interplay of cell-to-cell and cell-to-material interactions are incorporated. In this review, we summarize the recent developments in in vitro skin models, with a focus on their applications, limitations, and future directions.

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Section: Layer-by-layer Analysis Of Skin Tissue Modelsmentioning

confidence: 96%

Section: Basement Membrane Zonementioning

confidence: 99%

Layer-by-Layer Analysis of In Vitro Skin Models

Footner,

Firipis,

Liu

et al. 2023

ACS Biomater. Sci. Eng.

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“…The cytotoxicity of the fabricated hydrogels (ADA-Gel and ADA-Gel-PRP) was evaluated using a test on extracts followed by an MTT (3-[4-C-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay, as per ISO 10993-5 [17]. Briefly, the hydrogel discs, of a known size having a surface area of about 1.25 cm 2 , were incubating in 1 mL culture medium at 37 • C for 24 h. After incubation, the spent culture medium containing potential leachables from the hydrogel, or otherwise termed as extracts, were collected.…”

Section: % Hemolysis =mentioning

confidence: 99%

Formulation and Characterization of Alginate Dialdehyde, Gelatin, and Platelet-Rich Plasma-Based Bioink for Bioprinting Applications

et al. 2020

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Layer-by-layer additive manufacturing process has evolved into three-dimensional (3D) “bio-printing” as a means of constructing cell-laden functional tissue equivalents. The process typically involves the mixing of cells of interest with an appropriate hydrogel, termed as “bioink”, followed by printing and tissue maturation. An ideal bioink should have adequate mechanical, rheological, and biological features of the target tissues. However, native extracellular matrix (ECM) is made of an intricate milieu of soluble and non-soluble extracellular factors, and mimicking such a composition is challenging. To this end, here we report the formulation of a multi-component bioink composed of gelatin and alginate -based scaffolding material, as well as a platelet-rich plasma (PRP) suspension, which mimics the insoluble and soluble factors of native ECM respectively. Briefly, sodium alginate was subjected to controlled oxidation to yield alginate dialdehyde (ADA), and was mixed with gelatin and PRP in various volume ratios in the presence of borax. The formulation was systematically characterized for its gelation time, swelling, and water uptake, as well as its morphological, chemical, and rheological properties; furthermore, blood- and cytocompatibility were assessed as per ISO 10993 (International Organization for Standardization). Printability, shape fidelity, and cell-laden printing was evaluated using the RegenHU 3D Discovery bioprinter. The results indicated the successful development of ADA–gelatin–PRP based bioink for 3D bioprinting and biofabrication applications.

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“…SF can be fabricated into a variety of forms, viz. films, hydrogels, porous sponges, and micro/nanofibrous matrices. ,, Depending on the process, the parameters involved would strongly influence the properties of the resultant product. For instance, Tamada reported a new process of fabricating SF porous sponges by freeze-thawing of fibroin aqueous solution in the presence of a small amount of an organic solvent .…”

Section: Introductionmentioning

confidence: 99%

Optically Clear Silk Fibroin Films with Tunable Properties for Potential Corneal Tissue Engineering Applications: A Process–Property–Function Relationship Study

2022

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Owing to the shortage of donor corneas and issues associated with conventional corneal transplantation, corneal tissue engineering has emerged as a promising therapeutic alternative. Biocompatibility and other attractive features make silk fibroin a biomaterial of choice for corneal tissue engineering applications. The current study presents three modes of silk fibroin film fabrication by solvent casting with popular solvents, viz. aqueous (aq), formic acid (FA), and hexafluoroisopropanol (HFIP), followed by three standard modes of postfabrication annealing with water vapor, methanol vapor, and steam, and systematic characterization studies including corneal cell culture in vitro . The results indicated that silk fibroin films made from aq, FA, and HFIP solvents had surface roughness (Rq) of 1.39, 0.32, and 0.13, contact angles of 73°, 85°, and 89°, water uptake% of 58, 29, and 27%, swelling ratios of 1.58, 1.3, and 1.28, and water vapor transmission% of 39, 26, and 22%, respectively. The degradation rate was in the order of aq > HF > FA, whereas the tensile strength was in the order of aq < HF < FA. Further, the results of the annealing process indicated notable changes in morpho-topographical, physical, degradation, and tensile properties. However, the films showed no detectable changes in chemical composition and remained optically clear with >90% transmission in the visible range, irrespective of fabrication and postfabrication processing conditions. The films were noncytotoxic against L929 cells and were cytocompatible with rabbit cornea-derived SIRC cells in vitro . The study demonstrated the potential of fine-tuning various properties of silk fibroin films by varying the fabrication and postfabrication processing conditions.

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Fabrication of co-cultured tissue constructs using a dual cell seeding compatible cell culture insert with a clip-on scaffold for potential regenerative medicine and toxicological screening applications

Cited by 9 publications

References 37 publications

Layer-by-Layer Analysis of In Vitro Skin Models

Layer-by-Layer Analysis of In Vitro Skin Models

Formulation and Characterization of Alginate Dialdehyde, Gelatin, and Platelet-Rich Plasma-Based Bioink for Bioprinting Applications

Optically Clear Silk Fibroin Films with Tunable Properties for Potential Corneal Tissue Engineering Applications: A Process–Property–Function Relationship Study

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