Ontogenesis of the basement membrane zone after grafting cultured human epithelium: a morphologic and immunoelectron microscopic study.
Sheets of cultured epithelial cells have been successfully used as autografts for the permanent coverage of patients with full-thickness burns and as allografts to stimulate the healing of chronic skin ulcers. The basement membrane zone (BMZ), composed of lamina lucida, lamina densa, and anchoring fibrils, plays a pivotal role in the firm adherence of the epidermis to the dermis. The present study describes the ultrastructural development during various stages of wound healing after resurfacing different wound areas by cultured epithelial grafts. For this purpose, biopsies were obtained from five patients 5 d to 4.5 years after resurfacing full-thickness burns with cultured autografts, and from five patients at various stages after treatment of excised tattoos and chronic skin ulcers with cultured allografts. One biopsy was taken from a spontaneously healed burn wound 30 years post-injury. Ultrathin sections were prepared for transmission and immunoelectron microscopy, using monoclonal antibodies against type IV and VII collagen. Findings were compared to controls of age- and site-matched normal skin. Eleven days after grafting, the first BMZ features had regenerated, including lamina lucida, a discontinuous lamina densa, hemidesmosomes, and sparse anchoring fibrils. The process of de novo synthesis of BMZ components had begun, and within 4 to 5 weeks complete reformation of BMZ was observed, including normal distribution of anchoring fibrils. Immunolabeling of type VII collagen was first observed upon the lower part of lamina densa at day 11 and steadily increased, reaching normal values 5 weeks after grafting. In contrast, gold deposition of type IV collagen upon lamina densa was strongly increased at day 19 compared to normal. This high expression reduced a little at 5 weeks, but remained high up to 30 years after injury. Long-term burn scars exhibited pseudopodia-like extensions of all basal cells, abundant anchoring fibrils, and an increased amount of arching anchoring fibrils. These features might compensate for the lack of proper rete ridges.
Mommaas
A M
AM
Department of Dermatology, University Hospital Leiden, The Netherlands.
Teepe
R G
RG
Leigh
I M
IM
Mulder
A A
AA
Koebrugge
E J
EJ
Vermeer
B J
BJ
D000293:Adolescent; D000328:Adult; D001485:Basement Membrane / Q000254:growth & development*; D002056:Burns / Q000473:pathology; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D002921:Cicatrix / Q000473:pathology; D003094:Collagen / Q000737:chemistry; D006801:Humans; D007223:Infant; D016253:Microscopy, Immunoelectron; D008875:Middle Aged; D012867:Skin / Q000166:cytology / Q000648:ultrastructure; D016038:Skin Transplantation / Q000502:physiology*
D003094:Collagen