Evaluation of the Effects of Transforming Growth Factor–Beta 3 (TGF-β3) 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-β3) 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-β3 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-β3 (ChT); and (4) chitosan film containing TGF-β3–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 mm2) was similar to that of intact tendons (6.4 ± 1.1 mm2) ( 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-β3 had a positive effect on AT midsubstance healing, and the sustained-release NP form improved the outcomes, more specifically accelerating the remodeling process. Clinical Relevance: This study demonstrated the effectiveness of TGF-β3 on tendon healing on a rat model, which is an important step toward clinical use. The novel method of using PLGA-b-PEG NPs as a drug-delivery system with sustained-release properties had promising results.

Study of the Synergistic Immunomodulatory and Antifibrotic Effects of Dual-Loaded Budesonide and Serpine1 siRNA Lipid–Polymer Nanoparticles Targeting Macrophage Dysregulation in Tendinopathy

“…The strategy of silencing PAI-1 instead of the upstream mediator TGF-β1 is preferable since TGF-β1 has some beneficial effects on tendon healing, such as the induction of cell proliferation and ECM formation. 39 Therefore, certain downregulation of TGF-β1 can be beneficial, but complete silencing is not desirable. Never-Figure 2.…”

“…The expression of Serpine1 is significantly downregulated when RAW 264.7 cells and PMA-differentiated THP-1 cells pretreated with TGF-β1 are treated with siRNA@LPNs and dual-loaded LPNs for 24 and 48 h, compared to the positive controls (TGF-β1 and LPS + TGF-β1, respectively) (Figures D and D). The strategy of silencing PAI-1 instead of the upstream mediator TGF-β1 is preferable since TGF-β1 has some beneficial effects on tendon healing, such as the induction of cell proliferation and ECM formation . Therefore, certain downregulation of TGF-β1 can be beneficial, but complete silencing is not desirable.…”

“…The strategy of silencing PAI-1 instead of the upstream mediator TGF-β1 is preferable since TGF-β1 has some beneficial effects on tendon healing, such as the induction of cell proliferation and ECM formation. 39 Therefore, certain downregulation of TGF-β1 can be beneficial, but complete silencing is not desirable. Nevertheless, the silencing of its downstream mediator, PAI-1, allows for abolishing only the downside effect of TGF-β, which is the excessive formation of ECM in a disorganized manner.…”

An Atelocollagen Injection Enhances the Healing of Nonoperatively Treated Achilles Tendon Tears: An Experimental Study in Rats

“… 8 , 48 Previous studies have also demonstrated the potential effects of TGF-β3 on the tendinogenic differentiation of stem cells. 3 Recently, Cetik et al 8 reported the effectiveness of TGF-β3 on Achilles tendon healing in a rat model. The higher expression of these TGF-β isoforms in CG rats might have had a more favorable effect on tendon regeneration than that in NG rats.…”

“…TGF-b3 is well known for its antifibrotic effects. 8,48 Previous studies have also demonstrated the potential effects of TGF-b3 on the tendinogenic differentiation of stem cells. 3 Recently, Cetik et al 8 reported the effectiveness of TGF-b3 on Achilles tendon healing in a rat model.…”