1. TGF-β1 Improves Biomechanical Strength by Extracellular Matrix Accumulation Without Increasing the Number of Tenogenic Lineage Cells in a Rat Rotator Cuff Repair Model.
- Author
-
Arimura, Hitoshi, Tokunaga, Takuya, Karasugi, Tatsuki, Okamoto, Nobukazu, Taniwaki, Takuya, Mizuta, Hiroshi, Shukunami, Chisa, Sakamoto, Hidetoshi, and Hiraki, Yuji
- Subjects
HUMERUS physiology ,SCAPULA ,SUPRASPINATUS muscles ,ROTATOR cuff injuries ,ANIMAL experimentation ,BIOMECHANICS ,GENE expression ,IMMUNOHISTOCHEMISTRY ,IN situ hybridization ,POLYMERASE chain reaction ,RATS ,STATISTICAL sampling ,STAINS & staining (Microscopy) ,STATISTICS ,WOUND healing ,GENETIC markers ,PILOT projects ,STATISTICAL power analysis ,DATA analysis ,REVERSE transcriptase polymerase chain reaction ,DESCRIPTIVE statistics ,MANN Whitney U Test ,KRUSKAL-Wallis Test ,THERAPEUTICS ,PHYSIOLOGY - Abstract
Background: Transforming growth factor β1 (TGF-β1) positively regulates the tenogenic marker genes scleraxis (Scx) and tenomodulin (Tnmd) in mesenchymal progenitors in vitro. However, little is known about the effect of TGF-β1 on the expression of tenogenic markers during rotator cuff (RC) healing in rats. Hypothesis: TGF-β1 improves the biomechanical properties and histological maturity of reparative tissue in a rat RC repair model by stimulating the growth of tenogenic cells. Study Design: Controlled laboratory study. Methods: Adult male Sprague-Dawley rats (N = 180) underwent unilateral supraspinatus tendon-to-bone surgical repair and were randomly treated with a gelatin hydrogel presoaked in TGF-β1 (100 ng) or phosphate-buffered saline. The effects of TGF-β1 on RC healing were investigated at 2, 4, 6, 8, and 12 weeks postoperatively by immunostaining for proliferating cell nuclear antigen, by real-time reverse transcription polymerase chain reaction and in situ hybridization or immunostaining for enthesis-related markers (SRY-box containing gene 9 [Sox9], Scx, and Tnmd), and by real-time reverse transcription polymerase chain reaction and immunostaining for type I and III collagen. At 6 and 12 weeks postoperatively, biomechanical testing, micro–computed tomography, and biochemical analysis were also performed. At 2 and 4 weeks postoperatively, mesenchymal stem cell–related markers, phospho-Smad2, and matrix metalloproteinase 9 (MMP-9) and MMP-13 were assessed by immunostaining. Results: The TGF-β1-treated group had significantly higher ultimate load to failure and tissue volume at 6 and 12 weeks postoperatively and a higher collagen content at 12 weeks compared with the saline group. Tendon-related gene expression, histological maturity, cell proliferation, and mesenchymal stem cell–related marker immunoreactivity were not affected by exogenously administrated TGF-β1 at all time points. In the TGF-β1-treated group, the percentage of phospho-Smad2-positive cells within the healing tissue increased, whereas the expression of MMP-9 and MMP-13 significantly decreased at 2 and 4 weeks postoperatively. Conclusion: TGF-β1 enhances formation of tough fibrous tissues at the healing site by inhibiting MMP-9 and MMP-13 expression to increase collagen accumulation but without the growth of tenogenic lineage cells. Clinical Relevance: These findings suggest that TGF-β1 could be used for enhancing biomechanical strength after RC surgical repair. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF