Effects of perfusion and cyclic compression on in vitro tissue engineered meniscus implants.

ABSTRACT

PURPOSE: The purpose of this study was to investigate the influence of continuous perfusion and mechanical stimulation on bone marrow stromal cells seeded on a collagen meniscus implant. METHODS: Bone marrow aspirates from 6 donors were amplified in vitro. 10(6) human BMSC were distributed on a collagen meniscus implant. Scaffolds were cultured under static conditions (control) or placed into a bioreactor system where continuous perfusion (10 ml/min) or perfusion and mechanical stimulation (8 h of 10% cyclic compression at 0.5 Hz) were administered daily. After 24 h, 7 and 14 days, cell proliferation, synthesis of procollagen I and III peptide (PIP, PIIIP), histology, and the equilibrium modulus of the constructs were analyzed. RESULTS: Proliferation demonstrated a significant increase over time in all groups (p < 0.001). PIP synthesis was found to increase from 0.1 ± 0.0 U/ml/g protein after 24 h to 2.0 ± 0.5 (perfusion), 3.8 ± 0.3 (mechanical stimulation), and 1.8 ± 0.2 U/ml/g protein (static control, lower than perfusion and mechanical stimulation, p < 0.05). These differences were also evident after 2 weeks (2.7 ± 0.3, 4.0 ± 0.6, and 1.8 ± 0.2 U/ml/g protein, p < 0.01); PIIIP synthesis was found to increase from 0.1 ± 0.0 U/ml/g protein after 24 h to 2.9 ± 0.7 (perfusion), 3.1 ± 0.9 (mechanical stimulation), and 1.6 ± 0.3 U/ml/g protein (controls) after 1 week and remained significantly elevated under the influence of perfusion and mechanical stimulation (p < 0.01) after 2 weeks. Mechanical stimulation increased the equilibrium modulus more than static culture and perfusion after 2 weeks (24.7 ± 7.6; 12.3 ± 3.7; 15.4 ± 2.6 kPa; p < 0.02). CONCLUSION: Biomechanical stimulation and perfusion have impact on collagen scaffolds seeded with BMSCs. Cell proliferation can be enhanced using continuous perfusion and differentiation is fostered by mechanical stimulation.