Enhancing engineered ligament strength using a multi-strand approach.

Abstract

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 gel1. 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 ...