Allogeneic cultured dermal substitute composed of spongy collagen containing fibroblasts: evaluation in animal test

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Abstract
The authors developed a cultured dermal substitute (CDS) composed of a spongy collagen containing cultured fibroblasts. The cultured fibroblasts derived from Sprague Dawley rat skin were seeded on a spongy collagen at a density of 5 x 105 cells cm-2 and cultured for 7 days. This CDS was applied to the debrided wound of full-thickness burn which was inflicted experimentally on the dorsum of Wister rat, and then the wound conditions were observed over a period of 2 weeks. The comparative study was conducted using an acellular spongy collagen as well as a commercially available temporary wound dressing, Biobrane®, since a different type of cultured dermal substitute, Dermagraft-TC®, is composed of Biobrane®, whose inner site is combined with cultured fibroblasts. Each covering material was applied on the debrided wound area and exchanged by new one 1 week later. When the debrided wound was covered with Biobrane®, a small portion of necrotic tissue was observed I week after application, and the granulation tissue formation was greatly delayed. This wound area showed a poor granulation tissue even 2 weeks later. On the contrary, when covered with an acellular spongy collagen, no necrotic tissue was observed. This wound area showed a more or less irregular granulation tissue at 1 week and then a healthy granulation tissue 2 weeks later. This preliminary comparative study suggests that an acellular spongy collagen is able to function as a more suitable matrix for CDS, compared with Biobrane®. The wound area covered with a CDS assumed a moist, shiny, and hyperaemic appearance 1 week after application showing a healthy granulation tissue. The macroscopic evaluations indicate that the CDS is able to prepare a healthy granulation tissue at an earlier stage, compared with the acellular spongy collagen. In addition, the histologic views demonstrate that the CDS is able to prepare a thicker and denser granulation tissue, compared with the acellular spongy collagen. Although the fate of cultured fibroblasts in the CDS on the wound surface within 1 week is not clear, these findings suggest that fibroblasts in CDS are able to provide excellent conditions for wound healing.