Engineering of Uniform Epidermal Layers via Sacrificial Gelatin Bioink-Assisted 3D Extrusion Bioprinting of Skin

Minjun Ahn, Won Woo Cho, Hanju Lee, Wonbin Park, Seok Hyeon Lee, Jae Woo Back, Qiqi Gao, Ge Gao*, Dong Woo Cho*, Byoung Soo Kim*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

To reconstruct an ideal full-thickness skin model, basal keratinocytes must be distributed as a confluent monolayer on the dermis. However, the currently available extrusion bioprinting method for the skin is limited when producing an air-exposed cellular monolayer because the cells are encapsulated within a bioink. This is the first study to use sacrificial gelatin-assisted extrusion bioprinting to reproduce a uniform and stratified epidermal layer. Experimental analyses of the rheological properties, printability, cell viability, and initial keratinocyte adhesion shows that the optimal gelatin bioink concentration is 4 wt.%. The appropriate thickness of the bioprinted gelatin structure for achieving a confluent keratinocyte layer is determined to be 400 µm. The suggested strategy generates a uniform keratinocyte monolayer with tight junctions throughout the central and peripheral regions, whereas manual seeding generates non-uniform cellular aggregates and vacancies. These results influence gene expression, exhibiting a propensity for epidermal differentiation. Finally, the gelatin-assisted keratinocytes are bioprinted onto a dermis composed of gelatin methacryloyl and dermis-derived decellularized extracellular matrix to establish a full-thickness skin model. Thus, this strategy leads to significant improvements in epidermal differentiation/stratification. The findings demonstrate that the gelatin-assisted approach is advantageous for recreating reliable full-thickness skin models with significant consistency for mass production.

Original languageEnglish
Article number2301015
JournalAdvanced healthcare materials
Volume12
Issue number27
DOIs
Publication statusPublished - 27 Oct 2023

Keywords

  • confluent keratinocyte monolayers
  • extrusion bioprinting
  • sacrificial gelatin-based bioinks
  • skin engineering
  • stratified epidermal layers

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Ahn, M., Cho, W. W., Lee, H., Park, W., Lee, S. H., Back, J. W., Gao, Q., Gao, G., Cho, D. W., & Kim, B. S. (2023). Engineering of Uniform Epidermal Layers via Sacrificial Gelatin Bioink-Assisted 3D Extrusion Bioprinting of Skin. Advanced healthcare materials, 12(27), Article 2301015. https://doi.org/10.1002/adhm.202301015