Use Of Clotted Human Plasma And Aprotinin In Skin Tissue Engineering: A Novel Approach To Engineering Composite Skin On A Porous Scaffold | 30356
Journal of Tissue Science & Engineering
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Tissue-engineered composite skin is a promising therapy for treatment of chronic and acute wounds including burns. Providing
the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice
host cell infiltration and vascularisation to achieve permanent wound closure in a single-stage. However, the high porosity and
the lack of a supportive basement membrane in most commercially available dermal scaffoldshinders organized keratinocyte
proliferation and stratification in vitro and may delay re-epithelization in vivo. The objective of this study was to develop a method
to enable the in vitro production of a human skin equivalent (HSE) that included a porous scaffold and dermal and epidermal
cells expanded ex vivo with the potential to be used for definitive treatment of skin defects in a single procedure. A collagenglycosaminoglycan
dermal scaffold (IntegraŽ) was populated with adult fibroblasts. A near normal skin architecture was achieved
by addition of coagulated human plasma to the fibroblast-populated scaffold prior to seeding cultured keratinocytes. This resulted
in reducing scaffold pore size and improving contact surfaces. Skin architecture and basement membrane formation was further
improved by the addition of aprotinin (a serine protease inhibitor) to the culture media to inhibit premature clot digestion.
Histological assessment of the novel HSE revealed expression of keratin 14 and keratin 10 similar to native skin with a multilayered
neo-epidermis morphologically comparable to human skin. Furthermore, deposition of collagen IV and laminin-511 were
detected by immunofluorescence indicating the formation of a continuous basement membrane at the dermal-epidermal junction.
The proposed method was efficient in producing an in vitro near native HSE using the chosen ?off the shelf ? porous scaffold
(IntegraŽ). The same principles and promising outcomes should be applicable to other biodegradable porous scaffolds combined
with autologous cells for use in wound treatment.
Shiva Akbarzadeh obtained her PhD from the Ludwig Institute for Cancer Research, University of Melbourne in 2002. She has over 13 years of experience in cell
biology and signalling in a variety of systems. In 2009 she established the Skin Culture Laboratory at the Victorian Adult Burns Service- the Alfred, focusing on
translational research in skin regeneration. She leads the team responsible for manufacturing Cultured Epithelial Autograft for cellular based therapy of major burns
and research in developing novel skin substitutes for clinical application. In addition, she provides lab based training on skin tissue engineering for scientists and
surgeons from developing countries.
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