Epidermal Basement Membrane Substitutes for Bioengineering of Human Epidermal Equivalents

Nikola Kolundzic; Preeti Khurana; Debra Crumrine; Anna Celli; Theodora M Mauro; Dusko Ilic

doi:10.1016/j.xjidi.2021.100083

Epidermal Basement Membrane Substitutes for Bioengineering of Human Epidermal Equivalents

JID Innov. 2021 Dec 7;2(2):100083. doi: 10.1016/j.xjidi.2021.100083. eCollection 2022 Mar.

Authors

Nikola Kolundzic^{1

2}, Preeti Khurana^{1

2}, Debra Crumrine^{3

4}, Anna Celli^{3

4}, Theodora M Mauro^{3

4}, Dusko Ilic^{1

2}

Affiliations

¹ Department of Women & Children's Health, School of Life Course & Population Sciences, Faculty of Life Sciences & Medicine, King's College London, London, United Kingdom.
² Assisted Conception Unit (ACU), Guy's and St Thomas' NHS Foundation Trust, London, United Kingdom.
³ UCSF Department of Dermatology, University of California San Francisco, San Francisco, California, USA.
⁴ San Francisco Veteran Affairs Medical Center, San Francisco, California, USA.

Abstract

Epidermal basement membrane, a tightly packed network of extracellular matrix (ECM) components, is a source of physical, chemical, and biological factors required for the structural and functional homeostasis of the epidermis. Variations within the ECM create distinct environments, which can affect the property of cells in the basal layer of the epidermis and subsequently affect keratinocyte differentiation and stratification. Very little attention has been paid to mimicking basement membrane in organotypic cultures. In this study, using parameters outlined in a consensus on the quality standard of organotypic models suitable for dermatological research, we have evaluated three basement membrane substitutes. We compared fibronectin with three complex three-dimensional matrices: Matrigel, decellularized dermal fibroblast‒produced and ‒assembled ECM, and a dry human amniotic membrane. Our results suggest that Matrigel is not a suitable substrate for human epidermal equivalent culture, whereas the two other complex three-dimensional substitutes, decellularized dermal fibroblast‒produced and ‒assembled ECM and dry human amniotic membrane, were superior to single layer fibronectin coating. Human epidermal equivalents cultured on either decellularized dermal fibroblast‒produced and ‒assembled ECM or on dry human amniotic membrane generated hemidesmosomes, whereas those on fibronectin did not. In addition, human epidermal equivalent cultured on decellularized dermal fibroblast‒produced and ‒assembled ECM and on dry human amniotic membrane can be maintained in culture 4 days longer than human epidermal equivalent cultured on fibronectin without compromising the barrier function.

Keywords: 3D, three-dimensional; Ca++, calcium ion; ECM, extracellular matrix; HEE, human epidermal equivalent; HSE, human skin equivalent; K, keratin; KC, keratinocyte; TEER, transepithelial electrical resistance; dDF ECM, decellularized dermal fibroblast‒produced and ‒assembled extracellular matrix; dHAM, dry human amniotic membrane.