Effect of difference in fixation methods of tendon graft and the microfracture procedure on tendon-bone junction healing

Background: There are generally two methods of fixation for tendon grafts used in ligament reconstruction: bone tunnel fixation and anchor fixation. The microfracture (Mf) procedure is a technique to induce bleeding from the bone marrow, and the bleeding may contain cells with differentiation potential. However, few studies have compared the effects of the Mf procedure with those of the fixation methods. This study aimed to evaluate the effectiveness of the Mf procedure on two tendon graft fixation methods: histological, gene expression, tendon graft thickness, and mechanical. We especially focused our investigation on junction healing of tendon grafts and bone in the two fixation methods. Methods: We used 20 rabbits to evaluate tendon and bone healing in a peroneal tendon graft model. The rabbit models were divided into five groups according to the combination of peroneal tendon graft fixation method and Mf technique as follows: control group (C, n = 4), bone tunnel fixation without Mf procedure group (BT − Mf, n = 4), bone tunnel fixation with Mf procedure group (BT + Mf, n = 4), anchor fixation without Mf procedure group (A − Mf, n = 4), and anchor fixation with Mf procedure group (A + Mf, n = 4). All animals were sacrificed at 4 weeks postoperatively. The specimens underwent histological evaluation, mRNA analysis, tendon graft thickness at the tendon-bone junction, and biomechanical testing. Results: Histological evaluation of the BT + Mf and A + Mf groups showed healing with fibrocartilage formation at the tendon graft-bone junction. The mRNA expression showed significant increase in type 2 collagen, Scleraxis, and SRY-box9 in the BT + Mf and A + Mf groups. In biomechanical tests, the BT + Mf and A + Mf groups showed significantly increased tensile strength compared with the BT − Mf and A − Mf groups (BT + Mf group, 21.6 ± 1.7 N; A + Mf group, 22.5 ± 2.3 N vs. BT − Mf group, 12.3 ± 2.4 N; A − Mf group, 11 ± 2.3 N). Conclusion: The Mf procedure resulted in fibrocartilage formation at the tendon-bone junction in the BT and anchor fixation and improved the fixation strength at 4 weeks.