Development of cultured dermal substitute composed of hyaluronic acid and collagen spongy sheet containing fibroblasts and epidermal growth factor.
The present study aimed to develop a two-layered cultured dermal substitute (CDS). The upper layer is a hyaluronic acid (HA) and collagen (Col) spongy sheet with or without epidermal growth factor (EGF). The lower layer is a HA spongy sheet and Col gel containing fibroblasts. The CDS is prepared in serum-free medium, followed by placing on the wound surface. Corresponding to clinical application, CDS was incubated in serum-free medium for a period of 1, 3 or 5 days, followed by placing onto the air and culture medium interface (wound surface model), and culture for 6 days using conventional culture medium supplemented with serum. Metabolic activity and cytokine production were considerably higher in EGF-incorporating CDS, as compared with EGF-free CDS. Metabolic activity of EGF-incorporating CDS was maintained for a period of 3 days, but decreased slightly after 5 days. EGF-incorporating CDS is able to effectively stimulate fibroblasts within CDS to release increased amounts of vascular endothelial growth factor and hepatocyte growth factor, which are essential for wound healing. CDS is promising for wound therapy, because there is no risk of cellular damage caused by cryopreservation, thawing and rinsing processes. The critical issue is how to reduce the cellular damage during a prolonged period of incubation in serum-free medium. EGF-incorporating CDS can be used after a period of 3-5 days incubation in serum-free medium. This period is sufficient for transport of CDS from manufacturing facilities to hospitals.
['Kuroyanagi|Misato|M|', 'Yamamoto|Akiko|A|', 'Shimizu|Nahoko|N|', 'Ishihara|Eiko|E|', 'Ohno|Haruka|H|', 'Takeda|Akira|A|', 'Kuroyanagi|Yoshimitsu|Y|']
D000388:Air D018929:Cell Culture Techniques D003094:Collagen D003470:Culture Media D020405:Dermis D004815:Epidermal Growth Factor D005347:Fibroblasts D017228:Hepatocyte Growth Factor D006801:Humans D006820:Hyaluronic Acid D008855:Microscopy, Electron, Scanning D008954:Models, Biological D017768:Skin, Artificial D023822:Tissue Engineering D042461:Vascular Endothelial Growth Factor A D014945:Wound Healing D014947:Wounds and Injuries
hyaluronic acid collagen fibroblasts epidermal growth factor skin substitute