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TISSUE-ENGINEERED CARTILAGE: A MULTI-DIRECTIONAL ANALYSIS OF A GEL-BASED COMPOSITE



Abstract

The purpose of this study was to assess the physical, biochemical and biomechanical properties of a cartilage matrix-chondrocyte-fibrin glue composite as biological tool for cartilage repair.

Chondrocytes were enzymatically isolated from pig joints and resuspended in fibrinogen solution. Articular cartilage was harvested from pig joints, chopped into small chips and lyophiliaed. Cartilage chips were rehydrated and mixed with the cell/fibrinogen solution and with thrombin, in order to form a fibrin glue gel composite with cells and chips (group A). Control composites were made from lyophilised cartilage chips assembled with fibrin glue, but not containing chondrocytes (group B). Other control groups included fibrin glue/chondrocyte specimens without cartilage chips (group C) and specimens made of the fibrin glue alone (group D). All samples were weighed and implanted into subcutaneous pouches of nude mice. Animals were sacrificed at 2 and 9 weeks. Samples were evaluated grossly and the final/initial mass ratio was calculated. Samples were evaluated histologically, biomechanically, and biochemically.

Upon retrieval, only the samples in experimental group A retained their original pre-implantation mass. Histological analysis showed newly formed cartilage matrix in the specimens from group A and C. Biomechanical analysis showed significantly higher modulus in experimental samples, with respect to the other groups at the latest time point. Analysis of hydraulic permeability showed significantly decreasing values for all groups throughout the experimental times and lowest values for the experimental samples of group A in the latest time point, although there was no statistically significant difference among the groups. Biochemical analysis demonstrated higher values in the latest time point for samples prepared with cells for water and GAG content, whereas highest values for hydroxyproline were recorded for samples assembled with cartilage chips. DNA analysis showed higher values of samples prepared with chondrocytes and fibrin glue and also an important increase in values of the samples made of fibrin glue only, indicating a possible host fibroblast growth inside the samples over time. This tissue-engineered composite presents cartilaginous appearance and biomechanical integrity after 9 weeks in vivo.

NIH Grant R03AR45059 and SIOT Scholarship 2003