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A Novel Process for Optimizing Musculoskeletal Allograft Tissue to Improve Safety, Ultrastructural Properties, and Cell Infiltration

Whitlock, Patrick W. MD, PhD; Seyler, Thorsten M. MD; Parks, Griffith D. PhD; Ornelles, David A. PhD; Smith, Thomas L. PhD; Van Dyke, Mark E. PhD; Poehling, Gary G. MD

Journal of Bone & Joint Surgery - American Volume: 15 August 2012 - Volume 94 - Issue 16 - p 1458–1467
doi: 10.2106/JBJS.K.01397
Scientific Articles
Supplementary Content

Background: This study evaluated the properties of scaffold derived from freeze-dried human Achilles tendon allograft for use in anterior cruciate ligament (ACL) reconstruction. Our hypothesis was that such an allograft could be processed using a method to remove cellular and infectious material, producing a cytocompatible, architecturally modified scaffold possessing tensile properties suitable for ACL reconstruction.

Methods: Fifty-two allografts were provided by a tissue bank. Twenty-one were used as controls to assess cellularity, DNA content, microarchitecture, porosity, cytocompatibility, and tensile properties in vitro (n = 13) and in vivo (n = 8). Thirty-one were processed to produce scaffolds that were similarly assessed for these properties in vitro (n = 23) and in vivo (n = 8). The elimination of added enveloped and nonenveloped viruses was also determined in vitro after each processing step.

Results: A subjective decrease in cellularity and a significant decrease in DNA content were observed in the scaffolds compared with the allografts from which they had been derived. The porosity was increased significantly, and the scaffolds were cytocompatible in vitro. Processing resulted in significantly increased elongation of the scaffolds (138% of the elongation of the unprocessed allograft) during tensile testing. No other significant differences in tensile properties were observed in vitro or in vivo. The number of infiltrating host cells and the depth to which those cells infiltrated were significantly greater in the scaffolds. No enveloped viruses and only two of 108 nonenveloped viruses were detected in the scaffolds after processing, corresponding to a sterility assurance level of 0.2 × 10−7.

Conclusions: Allografts were processed using a method that removed cellular and infectious material to produce a decellularized, cytocompatible, architecturally modified scaffold with tensile properties that differed minimally from those of human allograft tissue both in vitro and in vivo. The scaffold production process also resulted in an increase in porosity that led to increased cell infiltration in vivo.

Clinical Relevance: Scaffolds derived from such tendon allografts have the potential to eliminate disease transmission and inflammation in recipients and to promote earlier and increased cell infiltration while retaining the initial tensile properties necessary to withstand rehabilitation after implantation.

1Department of Orthopaedic Surgery (P.W.W., T.M.S., T.L.S., and G.G.P.), Department of Microbiology and Immunology (G.D.P. and D.A.O.), and Institute for Regenerative Medicine (M.E.V.D.), Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157. E-mail address for P.W. Whitlock: pwhitloc@wfubmc.edu

Copyright 2012 by The Journal of Bone and Joint Surgery, Incorporated
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