Evaluation of the Effects of Transforming Growth Factor-Beta 3 (TGF-beta 3) Loaded Nanoparticles on Healing in a Rat Achilles Tendon Injury Model

Abstract

Background: Achilles tendon (AT) midsubstance injuries may heal suboptimally, especially in athletes. Transforming growth factor-beta 3 (TGF-beta 3) shows promise because of its recently discovered tendinogenic effects. Using poly(lactic-co-glycolic acid)-b-poly(ethylene glycol) (PLGA-b-PEG) nanoparticles (NPs) may enhance the results by a sustained-release effect. Hypothesis: The application of TGF-beta 3 will enhance AT midsubstance healing, and the NP form will achieve better outcomes. Study Design: Controlled laboratory study. Methods: A total of 80 rats underwent unilateral AT transection and were divided into 4 groups: (1) control (C); (2) empty chitosan film (Ch); (3) chitosan film containing free TGF-beta 3 (ChT); and (4) chitosan film containing TGF-beta 3-loaded NPs (ChN). The animals were sacrificed at 3 and 6 weeks. Tendons were evaluated for morphology (length and cross-sectional area [CSA]), biomechanics (maximum load, stress, stiffness, and elastic modulus), histology, immunohistochemical quantification (types I and III collagen [COL1 and COL3]), and gene expression (COL1A1, COL3A1, scleraxis, and tenomodulin). Results: Morphologically, at 3 weeks, ChT (15 +/- 2.7 mm) and ChN (15.6 +/- 1.6 mm) were shorter than C (17.6 +/- 1.8 mm) (P = .019 and = .004, respectively). At 6 weeks, the mean CSA of ChN (10.4 +/- 1.9 mm(2)) was similar to that of intact tendons (6.4 +/- 1.1 mm(2)) (P = .230), while the other groups were larger. Biomechanically, at 3 weeks, ChT (42.8 +/- 4.9 N) had a higher maximum load than C (27 +/- 9.1 N; P = .004) and Ch (29.2 +/- 5.7 N; P = .005). At 6 weeks, ChN (26.9 +/- 3.9 MPa) had similar maximum stress when compared with intact tendons (34.1 +/- 7.8 MPa) (P = .121); the other groups were significantly lower. Histologically, at 6 weeks, the mean Movin score of ChN (4.5 +/- 1.5) was lower than that of ChT (6.3 +/- 1.8). Immunohistochemically, ChN had higher COL3 (1.469 +/- 0.514) at 3 weeks and lower COL1 (1.129 +/- 0.368) at 6 weeks. COL1A1 gene expression was higher in ChT and ChN at 3 weeks, but COL3A1 gene expression was higher in ChN. Conclusion: The application of TGF-beta 3 had a positive effect on AT midsubstance healing, and the sustained-release NP form improved the outcomes, more specifically accelerating the remodeling process.