Flexor Tendon Tissue Engineering: Acellularization of Human Flexor Tendons with Preservation of Biomechanical Properties and Biocompatibility

• Published in: Tissue engineering. Part C, Methods Methods, 2011-08, Vol.17 (8), p.819-828
• Main Authors: Pridgen, Brian C., Woon, Colin Y.L., Kim, Maxwell, Thorfinn, Johan, Lindsey, Derek, Pham, Hung, Chang, James
• Format: Article
• Language: English
• Subjects: Biocompatible Materials - chemistry; Biomechanical Phenomena; Biomechanics; Biomedical materials; Cell Proliferation; Cell Survival; Cellular biology; DNA - metabolism; Fibroblasts - pathology; Glycosaminoglycans - chemistry; Humans; Physiological aspects; Skin - pathology; Tendon Injuries - pathology; Tendons; Tendons - pathology; Tensile Strength; Tissue engineering; Tissue Engineering - methods
• ISSN: 1937-3384; 1937-3392
• DOI: 10.1089/ten.tec.2010.0457

Objective: Acellular human tendons are a candidate scaffold for tissue engineering flexor tendons of the hand. This study compared acellularization methods and their compatibility with allogeneic human cells. Method: Human flexor tendons were pretreated with 0.1% ethylenediaminetetracetic acid (EDTA) for 4 h followed by 24 h treatments of 1% Triton X-100, 1% tri(n-butyl)phosphate, or 0.1% or 1% sodium dodecyl sulfate (SDS) in 0.1% EDTA. Outcomes were assessed histologically by hematoxylin and eosin and SYTO green fluorescent nucleic acid stains and biochemically by a QIAGEN DNeasy kit, Sircol collagen assay, and 1,9 dimethylmethylene blue glycosaminoglycan assay. Mechanical data were collected using a Materials Testing System to pull to failure tendons acellularized with 0.1% SDS. Acellularized tendons were re-seeded in a suspension of human dermal fibroblasts. Attachment of viable cells to acellularized tendon was assessed biochemically by a cell viability assay and histologically by a live/dead stain. Data are reported as mean±standard deviation. Result: Compared with the DNA content of fresh tendons (551±212 ng DNA/mg tendon), only SDS treatments significantly decreased DNA content (1% SDS [202.8±37.4 ng DNA/mg dry weight tendon]; 0.1% SDS [189±104 ng DNA/mg tendon]). These findings were confirmed by histology. There was no decrease in glycosaminoglycans or collagen following acellularization with SDS. There was no difference in the ultimate tensile stress (55.3±19.2 [fresh] vs. 51.5±6.9 [0.1% SDS] MPa). Re-seeded tendons demonstrated attachment of viable cells to the tendon surface using a viability assay and histology. Conclusion: Human flexor tendons were acellularized with 0.1% SDS in 0.1% EDTA for 24 h with preservation of mechanical properties. Preservation of collagen and glycoaminoglycans and re-seeding with human cells suggest that this scaffold is biocompatible. This will provide a promising scaffold for future human flexor tendon tissue engineering studies to further assess biocompatibility through cell proliferation and in vivo studies.