Human corneal epithelial equivalents constructed on Bombyx mori silk fibroin membranes

Bray, Laura J.; George, Karina A.; Ainscough, Louise; Hutmacher, Dietmar W.; Chirilă, Traian V.; Harkin, Damien G.

doi:10.1016/j.biomaterials.2011.03.068

Cited by 135 publications

(94 citation statements)

References 24 publications

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“…In addition, recent work has shown that both the protein structure and material properties of water-annealed silk films are consistent from batch to batch allowing for reproducible culture results as shown through various mechanical and biophysical testing methods 15,16 . In addition, the material's surface has shown great fidelity amongst film batches as indicated by SEM, atomic force micrscopy (AFM), and cell culture studies {Lawrence:2008wr, Omenetto:2008tc, Bray:2011kq} 7,23,24 . Material stability and consistency is an important factor to how the cell will sense the culture substrate through the various mechanotransduction pathways, and ultimately produce a desired/undesired cellular response 25,26 .…”

Section: Discussionmentioning

confidence: 99%

Silk Film Culture System for <em>in vitro</em> Analysis and Biomaterial Design

Lawrence

Pan

Weber

et al. 2012

JoVE

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Silk films are promising protein-based biomaterials that can be fabricated with high fidelity and economically within a research laboratory environment 1,2 . These materials are desirable because they possess highly controllable dimensional and material characteristics, are biocompatible and promote cell adhesion, can be modified through topographic patterning or by chemically altering the surface, and can be used as a depot for biologically active molecules for drug delivery related applications [3][4][5][6][7][8] . In addition, silk films are relatively straightforward to custom design, can be designed to dissolve within minutes or degrade over years in vitro or in vivo, and are produce with the added benefit of being transparent in nature and therefore highly suitable for imaging applications [9][10][11][12][13] . The culture system methodology presented here represents a scalable approach for rapid assessments of cell-silk film surface interactions. Of particular interest is the use of surface patterned silk films to study differences in cell proliferation and responses of cells for alignment 12,14 . The seeded cultures were cultured on both micro-patterned and flat silk film substrates, and then assessed through time-lapse phase-contrast imaging, scanning electron microscopy, and biochemical assessment of metabolic activity and nucleic acid content. In summary, the silk film in vitro culture system offers a customizable experimental setup suitable to the study of cell-surface interactions on a biomaterial substrate, which can then be optimized and then translated to in vivo models. Observations using the culture system presented here are currently being used to aid in applications ranging from basic cell interactions to medical device design, and thus are relevant to a broad range of biomedical fields. Video LinkThe video component of this article can be found at https://www.jove.com/video/3646/ Protocol Fabrication of Silicone Rubber Molds1. Produce or purchase a desired topographic surface for casting. For this publication a standard 100 mm etched silicon wafer will be described (Figure 1). 2. Weigh out polydimethylsiloxane (PDMS) potting (component A) and catalyst (component B) solution in a 1:9 ratio (9 g potting and 1 g catalyst) as provided in the purchased kit. 3. Mix solutions thoroughly to initiate curing process. 4. Place silicon wafer surface within a casting dish. 5. Weigh out 4.5 g of PDMS solution onto silicon wafer. 6. Spread PDMS solution as to cover a 100 mm diameter area of the wafer surface. 7. Tilt wafer to spread PDMS solution evenly. 8. Cover wafer with 100 mm diameter petri dish lid. 9. Place casting setup into 60 °C oven over night, making certain that curing is taking place on a flat surface. 10. Place cured PDMS/silicon wafer into 70% ethanol bath before removal. 11. Begin removing PDMS from wafer by using razor blade to lift edge (entire circumference) first. 12. Gently pull PDMS off using forceps within a 70% ethanol bath being careful not to tear silicone rubber casting....

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Section: Discussionmentioning

confidence: 99%

Silk Film Culture System for <em>in vitro</em> Analysis and Biomaterial Design

Lawrence

Pan

Weber

et al. 2012

JoVE

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“…44 Particularly, silk film topography significantly influenced initial human epithelial cell attachment, proliferation, and growth. 45,46 Further studies have shown that silk surfaces can be optimized by contact guidance to direct collective epithelial cell migration and the direction of tissue epithelialization. 47 Similarly, keratin-based substrates have been exploited for ocular surface reconstruction because of their good optical properties and ability to support epithelial cell growth in vitro.…”

Section: Corneal Epitheliummentioning

confidence: 99%

Corneal Tissue Engineering: Recent Advances and Future Perspectives

Ghezzi

Rnjak‐Kovacina

Kaplan

2015

Tissue Engineering Part B: Reviews

167

151

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“…While little of the native full-length protein remains following processing, the resulting peptide solutions can subsequently be used to generate a variety of stable and biocompatible materials including freestanding membranes, electro-spun fibers and porous sponges [24]. Of these three materials, the freestanding BMSF membranes have received most attention for potential use in corneal tissue reconstruction owing to their high degree of transparency and general handling properties [25][26][27][28]. Importantly, subsequent studies have revealed that BMSF-based materials support the attachment, growth and differentiation of cells isolated from all three main tissue layers of the cornea [25,[28][29][30].…”

Section: • Silk Fibroin Based Constructsmentioning

confidence: 99%

Recent advances in the design of artificial corneas

Griffith

Harkin

2014

Current Opinion in Ophthalmology

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Purpose of review: Artificial corneas are being developed to meet a shortage of donor corneas as well as to address cases where allografting is contraindicated. A range of artificial corneas has been developed. Here we review several newer designs and especially those inspired by naturally occurring biomaterials found with the human body and elsewhere. Summary:More recent attempts at making artificial corneas have taken a more nature-based or nature-inspired approach. Several will in the near future be likely to be available clinically.

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Human corneal epithelial equivalents constructed on Bombyx mori silk fibroin membranes

Cited by 135 publications

References 24 publications

Silk Film Culture System for <em>in vitro</em> Analysis and Biomaterial Design

Silk Film Culture System for <em>in vitro</em> Analysis and Biomaterial Design

Corneal Tissue Engineering: Recent Advances and Future Perspectives

Recent advances in the design of artificial corneas

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