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Am J Physiol Cell Physiol (February 27, 2002). doi:10.1152/ajpcell.00555.2001
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Articles in PresS, published online ahead of print February 6, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00555.2001
Submitted on December 3, 2001
Accepted on January 29, 2002

Fibroblast Collagen Secretion in Co-Culture with Keratinocytes: Normal Fibroblasts Secrete Collagen in a Keloid-Like Manner

Ivor J Lim1*, Toan-Thang Phan1, Boon-Huat Bay1, Robert Qi1, Hung-The Huynh1, Walter T Tan1, Seng-Teik Lee1, and Michael T Longaker1

1 Division of Plastic Surgery, Department of Surgery, National University of Singapore, Singapore, Singapore

* To whom correspondence should be addressed. E-mail: surlimi{at}nus.edu.sg.

Keloid scars represent a pathological response to cutaneous injury, reflecting a new set point between synthesis and degradation biased toward ECM collagen accumulation. Using a serum-free two-chamber co-culture model, we recently demonstrated a significant increase in normal fibroblast proliferation when co-cultured with keloid-derived keratinocytes. We hypothesized that similar keratinocyte-fibroblast interactions might influence fibroblast collagen production and examined conditioned media and cell lysate from co-culture for collagen I and III production by Western blot, allied with Northern analysis for procollagen I and III mRNA. Normal fibroblasts co-cultured with keloid keratinocytes produced increased soluble collagen I and III with a corresponding increase in pro-collagen I and III mRNA transcript levels. This was associated with decreased insoluble collagen from cell lysate. When keloid fibroblasts were co-cultured with keloid keratinocytes, both soluble and insoluble collagen were increased with associated procollagen III mRNA upregulation. Transmission electron microscopy of normal fibroblasts co-cultured with keloid keratinocytes showed an ECM appearance similar to in-vivo keloid tissue; an appearance not seen when normal fibroblasts were co-cultured with normal keratinocytes.




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