gms | German Medical Science

Introduction: Surgical resection of sarcomas may cause extensive soft tissue defects. Alternatively to plastic surgery a programmed filling with granular tissue with good biomechanical properties would be a desirable approach, in particular when seeded with autologous fibroblasts. An acellular dermal matrix (ADM) from human split-thickness skin was examined for its native structure, composition, cell- and DNA-freedom.

Materials and methods: To determine the collagen assembly and the structural preservation we visualized cryostat-cuts of ADM. ECM components were determined using immunofluorescence and CLSM. To determine the freedom of cells an immunofluorescence staining with antibodies against specific cell marker were performed. Presence of DNA was assessed qualitatively by histological analysis with propidium iodide and quantitatively by gel- electrophoresis for fragment length and by spectrophotometry for total DNA amount as well as by RT-PCR for GAPDH.

Results: Autofluorescence images showed the integrity of the natural structure of the collagen network with preserved gross structures like blood vessel canals. We could show that most of the ECM components were present. We detected a small amount of remaining cellwall debris but not intact cells. We were also able to show, that some measurable amounts of DNA had remained in the matrix as determined either by histological staining, gel electrophoresis or by spectrophotometry. However none of the performed RT-PCR for GAPDH was positive.

Conclusion: Processing of the ADM is not causing any damage to the integrity of the skin graft and does not degrade the quality of ECM proteins. Although the transplant is containing certain amounts of cell and DNA remnants, we could show that processing procedures lead to a total destruction of cells and active DNA. With no vital cells nor transcribable DNA it should be immunologically inert and due to it approved sterilisation method biologically save and it should be a suitable scaffold with potential for good revascularisation and wound integration.