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Enhancing engineered ligament strength using a multi‐strand approach.
Author(s) -
Paxton Jennifer,
Grover Liam
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.346.1
Subject(s) - petri dish , fibrin , ligament , biomedical engineering , tissue engineering , thrombin , in vivo , ascorbic acid , materials science , chemistry , anatomy , microbiology and biotechnology , biology , medicine , immunology , food science , platelet , genetics
Ligaments have a poor capacity for repair and options for developing engineered replacements are of significant interest. We have previously reported the creation of an engineered ligament using a cellular fibrin gel 1 . While these ligaments have a composition similar to native tissue, they can only withstand low loads under tension (~0.2N) and are, at present, too weak to withstand in vivo loads. This study aimed to investigate methods to augment the mechanical properties of engineered ligaments using a multi‐strand approach. Individual ligament constructs (LC) were made using 2 silk sutures pinned to a 35mm Sylgard‐coated Petri dish. A fibrin gel was made around the sutures using 50U/ml thrombin and 20mg/ml fibrinogen in growth media. 100K fibroblasts were seeded on the gel, incubated at 37°C and fed every 2‐3days with DMEM +250µM Ascorbic Acid (AA) and 50µM Proline (P). Formation of multi‐stand LC began after 3 weeks of formation of the original single‐stranded constructs (SS). SS were grouped together in 2s or 3s and placed side‐by‐side to make double‐(DS) or triple‐(TS) stranded LC (n=3) and fed as above. The tensile properties of the SS, DS and TS LC were assessed using an Instron microtester (10N load cell, 0.4mm/min rate). During culture, the individual strands in the DS and TS LC amalgamated into a distinct structure.The maximum load of the LC was augmented step‐wise by combining SS into DS or TS.Max. load was increased by 60.5% (SS‐DS, p=0.03), and by a further 71.4% (DS‐TS, p=0.001).LC type Single stranded (SS) Double stranded (DS) Triple stranded (TS)Maximum load (N) [mean (SD )] 0.28 (0.10) 0.45 (0.06) 0.77 (0.02)Our current work is focussed on increasing strand number per LC and assessing their histological characteristics. We would like to thank BBSRC and ORUK for funding.