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Seeding Keratinocytes on Papillary and Reticular Dermal Layers Remodels Skin Explants Differently: Towards Improved Skin Grafting

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Seeding Keratinocytes on Papillary and Reticular Dermal Layers Remodels Skin Explants DifferentlyTowards Improved Skin Grafting

Bage T, Eydmann T, Metcalfe A, Dheansa B, Mbundi L

Introduction

 

In full-thickness wounds such a severe burns, complex interactions between our cells and the extracellular matrixbecome disrupted.  Untreated, this leads to seriousclinical sequelae. 

 

Previous research in has highlighted differences between the effect of the papillary and reticular dermis in wound healing, suggesting that this could lead to possible optimisation1,2.

 

However, these studies were conducted in 3D skin models, not skin explants1,2.


Aims of the Study

 

1)To characterise the differences between the papillary and reticular dermal layers in supporting keratinocyte growth and re-epithelialisation
 
2)Assessthesesdifferencesinskinexplants, for thefirsttime
 
3)Consider any clinical implications of these differences


Methods

 

1.Use of a hand-held motorised dermatome to cut full-thickness (800µm) and partial-thickness (300-400µm) skin explants from human skin tissue
 
2.Inverted the explants to expose the papillary and reticular dermis without using enzymatic separation
 
3.Keratinocytes isolated from same donor seeded onto the exposed surface. At two weeks, sections were analysed histologically. 

 

Results

 

Cells seen on the surface of the explant confirmed to be keratinocytes with cytokeratin-14 staining (Image G)
 
Papillary sections seeded with keratinocytes (A, C, E) show basement membrane component formation by week two (Image C, yellow arrows show original tissue basement membrane laminin, green arrows show new laminin)
 
Reticular sections seededwith keratinocytes show increased proliferation and migration (not quantified) to reach complete dermal coverage by week two (Image B)
 
Seededkeratinocytesappeared to modulate their surrounding ECM to papillary-type (Images E&F, light blue staining)
 

All images 5x magnification

 

Discussion

Firstly, the results show that the two layers of the dermis modulate the repair process of skin in different ways. Since the principal cell type of the dermal compartments is fibroblasts, these findings suggest that the distinction between reticular and papillary fibroblasts is important in wound healing and repair.

 

In full-thickness wounds where the basement membrane has been compromised, the findings from this study suggest that papillary fibroblasts could be seeded in advance of a cultured epithelial autograft, to promote basement membrane reformation and support keratinocyte differentiation. In contrast, this study suggests that in cases where rapid complete wound coverage is a priority, such as in patients with high total burns surface area, perhaps mixed populations or reticular fibroblasts couldbe employed

 

This pilot study is not without its limitations, but it has highlighted exciting areas for further study.


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