Ka. Wright et al., ALTERNATIVE DELIVERY OF KERATINOCYTES USING A POLYURETHANE MEMBRANE AND THE IMPLICATIONS FOR ITS USE IN THE TREATMENT OF FULL-THICKNESS BURN INJURY, Burns, 24(1), 1998, pp. 7-17
Citations number
19
Categorie Soggetti
Dermatology & Venereal Diseases","Emergency Medicine & Critical Care
The Epicel ASAProgram(SM) service generates autologous keratinocyte gr
afts used for the closure of full-thickness wounds in moderately and s
everely burned patients. The manufacturing process used to generate Ep
icel(SM) service autografts (ESA) is based upon the keratinocyte co-cu
lture technique described by Rheinwald and Green which employs murine
Swiss 373/J2 fibroblasts as feeder cells. Recently, a technique has be
en described that employs a polyurethane wound dressing, HydroDerm(TM)
(HD, Innovative Technologies, Ltd), as a delivery vehicle for culture
d keratinocytes intended for autologous grafting. We have examined the
practical feasibility of this technique and report on testing the abi
lity of HD to support keratinocyte growth and epithelium formation in
vitro, at the air-liquid interface (ALI), and in vivo, after grafting
to full-thickness wounds created on the backs of athymic (Swiss Nu/Nu)
mice. The results demonstrate that keratinocytes grow on the HD dress
ing in Gibco SFM at a rate that is approximately 15 per cent of that o
bserved when cells are cultivated on tissue culture (TC) plastic using
standard techniques, yet the cells retain their proliferative capacit
y and form an epithelium in vitro when cultivated at the ALI on a derm
al substrate. Keratinocyte-seeded HD membranes were also transferred t
o full-thickness wounds in athymic mice. Animals grafted with cells se
eded to HD developed human epithelium, as revealed by species-specific
detection of involucrin and evolved a normal attachment to the wound
substratum, as demonstrated through the expression of dermally opposed
laminin and alpha 6 beta 4 integrin. The ability of keratinocytes to
maintain proliferative potential after seeding onto HD and their abili
ty to form a properly oriented epithelium in vitro and in vivo suggest
s that this wound dressing may be useful as a vehicle for autologous k
eratinocyte grafting and help to provide earlier epithelial coverage t
o the burned patient. However, because of the slow proliferation rate
of keratinocytes on HydroDerm, timely graft delivery would be best ach
ieved by combining cell expansion via the Rheinwald and Green culture
system, followed by the seeding of cells onto HydroDerm in a reduced c
alcium medium for subsequent autologous granting. (C) 1998 Elsevier Sc
ience Ltd for ISBI. All rights reserved.