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Plastic Surgery Research Council

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3d Printed Skin Substitute Using Human Dermal Fibroblasts And Human Epithelial Keratinoctyes
Jason Patel1, Liz Macias, PhD2, Joseph Willis1, Joel Berry, PhD3, Timothy W. King, MD, PhD, MSBE, FAAP, FACS1,2.
1UAB School of Medicine, Birmingham, AL, USA, 2UAB Department of Plastic Surgery, Birmingham, AL, USA, 3UAB Department of Biomedical Engineering, Birmingham, AL, USA.

PURPOSE:
Wound healing affects millions of people annually. Following injury, keratinocytes from the wound edge proliferate, migrate & differentiate to recapitulate the 3-D structure needed to provide a barrier function. If the wound is too large, skin grafting may be required. We are interested in discovering novel strategies to enhance the wound healing process. It may be possible to recreate a viable and histologically accurate skin tissue using three-dimensional (3D) printing with keratinocytes and dermal fibroblasts.
METHODS:
Adult Human Dermal Fibroblasts (HDFa) and Adult Human Epithelial Keratinocytes (HEKa; Cell Applications, San Diego, CA) were cultured and subsequently grown on Using a 3D bioprinter (Biobot, Advanced Solutions Life Sciences, Louisville, KY), a 3D printed hydrogel scaffold (Cellink, Boston, MA) was printed. The HEKa and HDFa cells were either seeded onto the 3D printed hydrogel scaffold or were incorporated into the hydrogel scaffold prior to 3D printing. Cell viability and histological appearance was determined by histologically sectioning the printed tissue and performing H&E as well as LIVE/DEAD staining (Thermo Fisher Scientific). The stained histological sections were analyzed for cell number, cell density, and tissue morphology.
RESULTS:
3D hydrogels were printed and seeded with HEKa and HDFa cells successfully. In addition, the HEKa and HDFa cells suspended in the hydrogel were successfully printed into 3D "skin" scaffolds. In both experimental groups the cells survived and were able to proliferate successfully.
CONCLUSION:
HEKa and HDFa cells can be grown on 3D printed hydrogels successfully. In addition, HEKa and HDFa cells can survive and grow when suspended in a hydrogel and 3D printed. Future potential applications of these results could lead to the creation of viable skin tissue for wound healing and surgical repair.


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