Burns 24 (1998) 104-113 Characterization of acellular dermal matrices (ADMs) prepared by two different methods Robert J. Waltera*, Takayoshi Matsudab, Hernan M. Reyes”, Jessica M. Walter”, Marella Hanumadassb ~‘Depurtment of Surgery, Division of Surgical Research. Cook County Hospital, Chicago, IL, USA “Department of Trauma, Bum Center; Cook Couny Hospital, Chicago, IL, USA Accepted 2 September 1997 Abstraet The efficacy of acellular dermal matrix (ADM) in the treatment of full-thickness skin injuries as a dermal substitute depends on its low antigenicity, capacity for rapid vascularization, and stability as a dermal template. These properties will be determined largely by the final composition of the ADM. We have treated human skin with either Dispase followed by Triton X-100 detergent or NaCl followed by SDS detergent, cryosectioned the resulting ADMs, and then characterized them immunohisto- chemically. Staining for cell-associated antigens (HLA-ABC, HLA-DR, vimentin, desmin, talin), extracellular matrix components (chondroitin sulfate, fibronectin, laminin, vitronectin, hyaluronic acid), elastin, and collagen type VII was dramatically reduced or absent from ADMs prepared by both methods. However, significant amounts of elastin, keratan sulfate, laminin, and collagen types III and IV were still observed in both ADMs. Both methods of ADM preparation resulted in extensive extraction of both cellular and extracellular components of the skin but retention of the basic dermal architecture. In general, ADM prepared by the NaCl-SDS method retained larger amounts of each antigen than did that prepared by the Dispase-Triton method. This was most evident for laminin and type VII collagen but larger amounts of type IV collagen, fibronectin, desmin, elastin, and HLA-DR were also evident in the NaCl-SDS ADM. 0 1998 Published by Elsevier Science Ltd for ISBI. All rights reserved. 1. Introduction The dermis provides stability to split-thickness skin grafts (STSGs) and to cultured epidermal autografts (CEAs). A sufficiently thick dermal bed at the recip- ient site inhibits contraction and hypertrophic scar formation of STSGs [l] and improves the survival of CEAs [2,3]. In recent years, many attempts have been made to produce a dermal substitute capable of supporting thin. STSG or CEA [1,4-lo], but none have proven entirely satisfactory. The most acceptable substitutes produced thus far appear to be those which employ an ac’ellular dermal matrix (ADM) derived from full- or split-thickness skin treated to remove epithelial components (keratinocytes, sweat glands and sebaceous glands) and dermal components (fibroblasts, vascular endothelium and smooth muscle) [8,9,11-131. “Correspondence should be addressed to: Dr Robert J. Walter. Department of Surgery, Hektoen Institute for Medical Research, Cook County Hospital, 627 South Wood Street, Chicago, IL 60612, USA. Tel.: +(312) 633-7237; Fax: +(312) 633-8347. These dermal substitutes must exhibit three important properties; very low antigenicity, the capacity for rapid vascularization, and stability as a dermal template. Earlier studies employed several methods for producing ADM from skin including treatment with trypsin [11,14,15], freeze-thawing [8,12,16], and lengthy incubations during which time endogenous enzymes act upon the dermis [9,13,1’7]. For the most part, ADMs produced by these methods have proven to be too highly antigenic when implanted into recipi- ents where they induce immune reaction resulting in poor graft survival [11,12,14,15]. Recently, using more effective and controlled extraction methods, ADMs exhibiting very low antigenicity, excellent stability, and retaining native dermal structure have been produced. Such ADMs have been produced using hypertonic NaCl followed by sodium dodecyl sulfate (NaCl-SDS ADM) and freeze-drying (AlloDenm) [lg. 191 or by using sequential treatments with Dispase followed by Triton X-100 (Dispase-Triton ADM) [20-221. To use these ADMs most effectively in wound management and to further develop composite skin 0305-4179/98/$19.00 + 0.00 0 1998 Published by Elsevier Science Ltd for ISBI. All rights reserved. PII: s0305-4179(97)00110-1