Application of PLGA-collagen hybrid mesh for three-dimensional culture of canine anterior cruciate ligament cells

Canine anterior cruciate ligament (ACL) cells were isolated from the anterior cruciate ligament and subcultured in DMEM containing 10% FBS. After being subcultured twice, the ACL cells were seeded in a PLGA-collagen hybrid mesh and cultured in DMEM containing 10% FBS in vitro for 16 days. After 1 day, the cells adhered and spread well on collagen microsponges of the hybrid mesh. The cells proliferated and excreted extracellular matrices with culture time. After 16 days of culture, the cells and extracellular matrices occupied the spaces in the scaffold. At that point, the cell/mesh constructs were rolled up and implanted subcutaneously in the dorsum of nude mouse. The implants were harvested after 2, 4, 8, and 12 weeks of implantation and examined by histological staining. The implants were macroscopically white. Histological examination of cross sections of the implant by hematoxylin and eosin staining showed a large population of cells and newly deposited collagen in the rolled implants seeded with ACL cells. The cells were surrounded with bundles of collagen fibers. The layers in the rolled implant became integrated after 8 weeks. The polymer fibers could still be seen 8 weeks after implantation and gradually degraded and eventually disappeared after 12 weeks.