Ex vivo culture of keratinocytes on papillary and reticular dermal layers remodels skin explants differently: towards improved wound care

Abstract

In this study, we characterised the effect that seeding keratinocytes on the papillary and reticular dermis had on the extracellular matrix and tissue integrity ex vivo. Human skin explants from consented patients (n = 6) undergoing routine surgery were cultured at a liquid–air interface, dermal-side up, and autologous keratinocytes seeded on the exposed papillary or reticular layer. After 7–21 days, histological and immunohistochemical evaluation of the morphology and extracellular matrix was performed. While the dermis remained robust in all explants cultures, keratinocytes seeded on the papillary layer showed less tissue infiltration and remodelling and formed clusters across the tissue. In contrast, keratinocytes seeded on the reticular layer infiltrated the tissue homogenously with an intact single-cell-layer surface coverage and structural changes characterised by increased deposition of ground substance, glycosaminoglycans, and collagen VII in 14 days. In addition, while the papillary section showed more new laminin deposition by 14 days than the reticular section, the latter expressed more connexin 43. These differences in re-epithelialisation and extracellular matrix characteristics suggest that wound depth and graft thickness may play a key role in wound healing and indicate that ECM characteristics should be factored in when designing biomaterials for wound applications and in the selection of recipient sites when using cells for grafting.

Development of Chitosan/Silver Sulfadiazine/Zeolite Composite Films for Wound Dressing

“…A burn is a wound of any traumatic type that compromises the function of the epithelial tissue. It is considered a significant problem, not only for the gravity when acute, but also concerning its significant sequelae that may forever mark burned patients [1]. This type of injury is distinguished from others due to the high risk of colonization by pathogenic bacteria, the presence of large amounts of non-viable tissue, the loss of a large quantity of water and blood, risk to remain open for extended periods of time until its complete healing, and frequently need to mobilize tissue for wound closure [2,3].…”

Elastin‐Like Recombinamers: Deconstructing and Recapitulating the Functionality of Extracellular Matrix Proteins Using Recombinant Protein Polymers

“…[16] Considering natural ECM heterogeneity and its importance to many fundamental cellular process, numerous tissue culture techniques, in vivo studies, and biomaterials have been developed in regenerative medicine to understand the interplay between ECM and cells so as to elucidate on and harness these qualities for biomedical applications. [15,47] In this regard, researchers using in vitro monolayer cultures of human cells have relied on coating tissue culture dishes (mostly plastic or glass) with different purified ECM-derived molecules so as to understand cell-ECM interactions. [48,49] To address the absence of the multidimensional 3D environment physicochemical cues, purified preparations or a mixture of ECM proteins (e.g., collagen, fibrinogen, elastin, and silk) has been formulated into 3D scaffolds with promising results both in vitro and in vivo for different applications.…”