A study of cytokines released from fibroblasts in cultured dermal substitute

Allogeneic cultured dermal substitute (CDS) was prepared by culturing fibroblasts on a two-layered spongy matrix of hyaluronic acid ( HA) and atelo-collagen (Col). CDS can be cryopreserved and transported to other hospitals in a frozen state. The present study was designed to analyze amounts of cytokines released from fibroblasts in fresh or cryopreserved CDS. The culture medium used in preparing CDS over a cultivation period of 1 week ( fresh CDS culture medium sample) contained vascular endothelial growth factor ( VEGF), basic fibroblast growth factor ( bFGF), hepatocyte growth factor (HGF), platelet derived growth factor ( PDGF)-AA, transforming growth factor (TGF)-beta 1, keratinocyte growth factor (KGF), interleukin (IL)-6 and IL-8. After thawing of cryopreserved CDS, the CDS was re-cultured in medium for 1 week. The culture medium used in re-culturing CDS for 1 week ( cryopreserved CDS culture medium sample) contained VEGF, bFGF, and HGF in the same concentration as before freezing, and TGF-beta 1 and IL-8 at half the concentration before freezing. Levels of PDGF-AA, KGF, and IL-6 were significantly less than before freezing. This finding suggests that the cryopreserved CDS retains its ability to release VEGF, bFGF, and HGF that are essential for wound healing.