TRITON-X is most effective among three decellularization agents for ACL tissue engineering.
Ruptures of the anterior cruciate ligament (ACL) are still associated with high rates of long-term complications, even in patients undergoing modern, state-of-the-art replacement. Tissue-engineering approaches have been shown to be of value in improving treatment of ACL ruptures. However, the success of tissue-engineering procedures depends on the choice of an appropriate biomaterial. Decellularized ACL tissue potentially combines the structural composition of the targeted tissue with a reduced risk of graft rejection or disease transmission. In this study, we tested the effectiveness of currently available decellularization methods based on TRITON-X, sodium dodecyl sulfate (SDS), and trypsin. After identifying the most effective decellularization method, the capacity for reseeding with ACL fibroblasts was studied. All decellularization protocols reduced DNA content, with TRITON-X treatment having the greatest effect. Concurrently, decellularization did not affect tissue collagen or total protein content, but did decrease glycosaminoglycan content. TRITON-X also resulted the least glycosaminoglycan depletion. Porcine ACL tissue after decellularization with TRITON-X could be successfully reseeded with human ACL fibroblasts as demonstrated by steady DNA content and increasing pro-collagen expression.
Vavken
Patrick
P
Sports Medicine Research Laboratory, Department of Orthopedic Surgery, Children's Hospital Boston, Harvard Medical School, 300 Longwood Avenue, Enders 1016, Boston, Massachusetts 02115, USA. patrick.vavken@childrens.harvard.edu
Joshi
Shilpa
S
Murray
Martha M
MM
D000818:Animals; D016118:Anterior Cruciate Ligament / Q000166:cytology / Q000187:drug effects* / Q000637:transplantation; D016025:Bone Transplantation / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000032:analysis; D004247:DNA / Q000032:analysis; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D008297:Male; D011092:Polyethylene Glycols / Q000737:chemistry / Q000494:pharmacology*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013501:Surface-Active Agents / Q000737:chemistry / Q000494:pharmacology*; D013552:Swine / Q000502:physiology*; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry; D014184:Transplantation, Homologous
D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D003094:Collagen; D004247:DNA