Assembly of bone marrow stromal cell sheets with knitted poly (L-lactide) scaffold for engineering ligament analogs

The current cell seeding technique has several disadvantages, such as low efficiency of cell attachment to scaffolds and the limited strength of cell-gel composite adhesion to scaffold. These problems warrant further study to improve the assembly of cell to scaffold. Therefore this study aims to fabricate a bone marrow stromal cells (bMSCs) sheet and assemble it on a knitted poly (L-lactide) (PLLA) scaffold for engineering ligament analogs. bMSCs were cultured to form a cell sheet in the presence of ascorbic acid. Once a sheet of bMSCs was obtained, it was assembled onto the knitted scaffold by a wrapping technique. Then the assembled structure was held in place in a spinner flask for 4 weeks. The macromorphology, histology, and biomechanics of the grafts were evaluated. The composite of cell sheet/PLLA scaffold constructs had transformed into tissuelike ligament analogs. Immunohistochemical analysis showed that the components of the analogs were similar to that of ligament tissues, consisting primarily of collagen type I and small amount of collagen type III and tenascin. The failure force of the cell/scaffold assembly under tension (46.68+/-2.29 N) was higher than that of the scaffold group (43.58+/-2.41 N; p<0.05), but tensile stiffness of the cell/scaffold group (20.6+/-1.417 N/mm) was significantly lower than that of the scaffold group (27.6+/-1.449 N/mm; p<0.05). These data showed that the incorporation of bMSCs sheet onto the PLLA scaffold could make the analog stronger and more stretchable. Therefore the approach of assembling bMSCs sheet onto knitted PLLA scaffold is promising for producing tissuelike and functional ligament analogs under dynamic fluid situation for the purpose of anterior cruciate ligament (ACL) reconstruction.