[A PRELIMINARY STUDY ON SMALL INTESTINAL SUBMUCOSA-SILK COMPOSITE SCAFFOLD TO RECONSTRUCT ANTERIOR CRUCIATE LIGAMENT].

OBJECTIVE: To prepare the small intestinal submucosa (SIS)-silk composite scaffold for anterior cruciate ligament (ACL) reconstruction, and to evaluate its properties of biomechanics, biocompatibility, and the influence on synovial fluid leaking into tibia tunnel so as to provide a better choice in the clinical application of ACL reconstruction.

METHODS: The silk was used to remove sericin and then weaved as silk scaffold, which was surrounded cylindrically by SIS to prepare a composite scaffold. The property of biomechanics was evaluated by biomechanical testing system. The cell biocompatibility of scaffolds was evaluated by live/dead staining and the cell counting kit 8 (CCK- 8). Thirty 6-week-old Sprague Dawley rats were randomly assigned to 2 groups (n = 15). The silk scaffold (S group) and composite scaffold (SS group) were subcutaneously implanted. At 2, 4, and 8 weeks after implanted, the specimen were harvested for HE staining to observe the biocompatibility. Another 20 28-week-old New Zealand white rabbits were randomly assigned to the S group and SS group (n = 20), and the silk scaffold and composite scaffold were used for ACL reconstruction respectively in 2 groups. Furthermore, a bone window was made on the tibia tunnel. At last, the electric resistance of tendon graft in the bone window was measured and recorded at different time points after 5 mL of 10% NaCl or 5 mL of ink solution was irrigated into the joint cavity recspectively.

RESULTS: The gross observation showed that the composite scaffold consisted of the helical silk bundle inside which was surrounded by SIS. The maximal load of silk scaffold and composite scaffold was respectively (138.62 ± 11.41) N and (137.05± 16.95) N, showing no significant difference (P > 0.05); the stiffness was respectively (24.65 ± 2.62) N/mm and (24.21 ± 2.39) N/mm, showing no significant difference (P > 0.05). The live/dead staining showed that the cells had good activity on both scaffolds. However, the cells on the composite scaffold had better extensibility. In addition, the cell proliferation curve indicated that no significant difference in the absorbance (A) values was founded between groups at various time points (P > 0.05). HE staining showed less inflammatory cells and much more angiogenesis in SS group than in S group at 2, 4, and 8 weeks after subcutaneously implanted (P < 0.05), indicating good biocompatibility. Additionally, the starting time points of electric resistance decrease and the ink leakage were both significantly later in SS group than in S group (P < 0.05). The duration of ink leakage was significantly longer in SS group than in S group (P < 0.05).

CONCLUSION: The SIS-silk composite scaffold has excellent biomechanical properties and biocompatibility and early vacularization after in viva implantation. Moreover, it can reducing the leakage of synovial fluid into tibia tunnel at the early stage of ACE reconstruction. So it is promising to be an ideal ACL scaffold.