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The MTT assay showed that the cell proliferation on hydroxyapatite (HAp) and HAp/bone morphogenic protein (BMP) coated group was better than the control and BMP coated groups at 5 days. And after 7 days of culture, the mRNA expression levels of type I collagen, osteonectin, osteopontin, bonesialoprotein, BMP-2, alkaline phosphatase (ALP) and Runx-2 in the HAp/BMP coated group were significantly higher than the other groups. Also, in this group showed the most significant induction of osteogenic gene expression compared to mesenchymal stem cells (MSCs) grown on the other groups. In addition, the cells in the HAp/BMP coated group delivered higher levels of ALP than the other three groups. Also, silk scaffolds were implanted as artificial ligaments in knees of rabbits, and they were harvested 1 and 3 months after implantation. On gross examination, HE staining showed that new bone tissue formation was more observed in the HAp/BMP coated group 3 weeks postoperatively. And masson staining showed that in the HAp/BMP coated group, the silk fibers were encircled by osteoblast, chondrocyte, and collagen. Furthermore, the analysis showed that the width of the graft-bone interface in the HAp and HAp/BMP coated group was narrower than that in the other two groups 3 weeks postoperatively. So, it is concluded that BMP incorporated HAp coated silk scaffold can be enhanced osseointegration and osteogenesis in bone tunnel. As a result, these experimental designs have been demonstrated to be effective in the acceleration of graft-to-bone healing by increasing new bone or fibrocartilage formation at the interface between graft and bone.

Enhancement of osseointegration of artificial ligament by nano-hydroxyapatite and bone morphogenic protein-2 into the rabbit femur