Regional variability, processing methods, and biophysical properties of human fascia lata extracellular matrix.

This study aims to assess the regional variability, processing methods, mechanical, biochemical, and cellular properties of human fascia lata as a scaffold for soft tissue repair and tissue engineering applications. Ten pairs of fascia lata (donor age 18-55) were used. One fascia patch from each pair was used to assess the geometric and biomechanical variability of fresh fascia. The other from each pair was subjected to 1 of 2 allograft processing methods: antibiotic soak alone or acellularization plus antibiotic soak. Stiffness, modulus, hydroxyproline, chondroitin/dermatan sulfate glycosaminoglycan (CSDS GAG), and DNA content were quantified in fascia from fresh and treated groups. The effect of location was not significant for thickness or stiffness within a 6 x 12 cm2 region of the iliotibial tract of fresh human fascia lata. Processing did not significantly change the stiffness, modulus, or CSDS GAG content of fascia ECM. However, hydroxyproline (collagen) content is significantly reduced in acellularized fascia, probably reflecting a removal of soluble collagen during the treatment (p < 0.02). Processing reduced the DNA content of fresh fascia approximately 10-fold (p < 0.001). The mechanical, chemical and ultrastructural similarities between fascia lata and tendon may make fresh or processed fascia an attractive ECM scaffold for soft tissue, particularly tendon, repair.

Mechanical conditioning of cell-seeded small intestine submucosa: a potential tissue-engineering strategy for tendon repair.

Our long-term objective is to enhance tendon repair by delivering cells on natural biologic scaffolds to the repair site. Clinical outcomes may be improved by first preconditioning these cell-seeded constructs in bioreactors to enhance their properties at implantation and to deliver cells expressing a desired phenotype. In this work, we have investigated the effect of in vitro mechanical conditioning on small-intestine submucosa (SIS) scaffolds seeded with primary tendon cells (tenocytes). SIS scaffolds (with and without cells) were conditioned under various loading regimes over a 2-week period. In vitro cyclic loading significantly increased the biomechanical properties (e.g., stiffness) of cell-seeded SIS constructs (129.1 +/- 10.2%) from time 0. The stiffness change of cyclically loaded constructs without cells was 33.9 +/- 13.8% and of statically loaded constructs with cells was 34.0 +/- 15.2% and without cells was 33.4 +/- 10.7%. In the cell-seeded groups, our data demonstrate a direct role (e.g., cell tensioning) for cells in construct stiffening. In addition, the initial stiffness of the cell-seeded, cyclically loaded constructs was found to be a strong predictor of the change in construct stiffness. Despite the mechanical integrity of these constructs being significantly less than native tendon, our data show that structural properties can be improved with in vitro mechanical conditioning. These data provide the basis for future studies investigating in vitro conditioning (mechanical, chemical) of cell-seeded ECM scaffolds and the use of such constructs for enhancing tendon repair in vivo.

Commercial extracellular matrix scaffolds for rotator cuff tendon repair. Biomechanical, biochemical, and cellular properties.

BACKGROUND: We are not aware of any in vitro study comparing the biomechanical, biochemical, and cellular properties of commercial extracellular matrix materials marketed for rotator cuff tendon repair. In this study, the properties of GraftJacket, TissueMend, Restore, and CuffPatch were quantified and compared with each other. The elastic moduli were also compared with that of normal canine infraspinatus tendon. METHODS: Samples were tested from different manufacturing lots of four materials: GraftJacket (ten lots), TissueMend (six), Restore (ten), and CuffPatch (six). The Kruskal-Wallis test was used to compare thickness, stiffness, and modulus as well as hydroxyproline, chondroitin/dermatan sulfate glycosaminoglycan, hyaluronan, and DNA contents among these matrices. The moduli of the extracellular matrices were also compared with those of normal canine infraspinatus tendon. RESULTS: All four extracellular matrices required 10% to 30% stretch before they began to carry substantial load. Their maximum moduli were realized in their linear region at 30% to 80% strain. The elastic moduli of all four commercial matrices were an order of magnitude lower than that of canine infraspinatus tendon. TissueMend had significantly higher DNA content than the other three matrices (p<0.0001), although both Restore and GraftJacket also had measurable amounts of DNA. CONCLUSIONS: Our data demonstrate chemical and mechanical differences among the four commercial extracellular matrices that we evaluated. Probably, the source (dermis or small intestine submucosa), species (human, porcine, or bovine), age of the donor (fetal or adult), and processing of these matrices all contribute to the unique biophysical properties of the delivered product. The biochemical composition of commercial extracellular matrices is similar to that of tendon. However, the elastic moduli of these materials are an order of magnitude lower than that of tendon, suggesting a limited mechanical role in augmentation of tendon repair.