Hyaluronic Acid Accelerates Tendon-to-Bone Healing After Rotator Cuff Repair

Background: There is growing evidence that the subacromial injection of hyaluronic acid (HA) is effective for pain relief in rotator cuff tears; however, its effect on tendon-to-bone healing remains unknown. Purpose: To examine the effect of HA on the chondrogenesis of mesenchymal stem cells (MSCs) in vitro and on tendon-to-bone healing in a rotator cuff repair model. Study Design: Controlled laboratory study. Methods: Bilateral complete tears of the infraspinatus tendon were made in rabbits and subsequently repaired. Before closure, 1 mL HA was applied to the repaired site, and phosphate-buffered saline was used in the opposite side as a control. Biomechanical, histological, and immunohistochemical analyses were performed at 4, 8, and 12 weeks after surgery. After euthanizing each animal, the bone marrow was isolated from the femoral bone in the same rabbits. Then, MSCs were cultured in media for chondrogenic differentiation, and the chondral pellet production and cartilage-related gene expression levels in the cells were examined at various concentrations of HA. Results: At 4 and 8 weeks after surgery, ultimate load-to-failure was significantly greater in the HA group than in the control group (45.61 +/- 9.0 N vs 32.42 +/- 9.4 N at 4 weeks, 90.7 +/- 16.0 N vs 66.97 +/- 10.0 N at 8 weeks; both P < .05) but not at 12 weeks after surgery (109.6 +/- 40.2 N vs 108.1 +/- 42.6 N, P > .05). Linear stiffness was not significant throughout the time point evaluation. The chondroid formation area at the tendon-bone interface stained by safranin O (control vs HA group) was 0.33% +/- 0.7% versus 13.5% +/- 12.3% at 4 weeks after surgery (P < .05) and 3.0% +/- 5.9% versus 12.9% +/- 12.9% at 8 weeks after surgery (P < .05), but there was no significant difference at 12 weeks after surgery. Maturity of collagen at the repaired site stained by PicroSirius Red (control vs HA group) was 16.2 +/- 10.6 versus 43.5 +/- 21.3 at 4 weeks after surgery (P < .05), but there were no significant differences at 8 and 12 weeks after surgery. MSCs were cultured in media for chondrogenic differentiation, and the chondral pellet production and cartilage-related gene expression levels in the cells were examined at various concentrations of HA. The number of CD44-positive cells (control vs HA group) was 8.3% +/- 1.4% versus 26.2% +/- 5.2% at 3 days after surgery (P < .05), 1.8% +/- 1.1% versus 26.6% +/- 11.6% at 4 weeks after surgery (P < .05), 0.6% +/- 0.9% versus 0.5% +/- 0.6% at 8 weeks after surgery (P > .05), and 1.8% +/- 4.0% versus 5.4% +/- 4.2% at 12 weeks after surgery (P > .05). Compared with the control group, HA significantly increased the volume of cartilaginous pellet produced by MSCs (0.0016 +/- 0.0015 mm(3) at 0 mg/mL of HA, 0.0041 +/- 0.0023 mm(3) at 1.0 mg/mL, and 0.0041 +/- 0.0018 mm(3) at 4.0 mg/mL), with increased mRNA expression (relative ratio to control) of type 2 collagen (1.34 +/- 0.38), SOX9 (1.58 +/- 0.31), and aggrecan (1.30 +/- 0.22) genes in the pellet (P < .01). Conclusion: HA accelerated tendon-to-bone healing in the rotator cuff repair model, enhancing the biomechanical strength and increasing chondroid formation and tendon maturity at the tendon-bone interface. Based on the data of in vitro experiments, HA-activated MSCs may play a crucial role in the acceleration of tendon-to-bone healing.