Rat Collagen Type III ELISA Kit

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is essential for maintaining normal function of the adult heart and is known to play an important role in myocardial remodeling. In the present study, we observed that heart-specific HB-EGF transgenic (TG) mice had systolic dysfunction with decreased fractional shortening (FS%), increased end-systolic diameter (LVIDs) at 5 months of age, increased heart fibrosis, and increased mRNA expression of Col1 alpha 1 and Col3 alpha 1 at 1, 3, 5 and 7 months of age compared to nontransgenic (NTG) littermates. However, the left ventricular anterior wall thickness at end-systole (LVAWs) of the TG mice was not different than the NTG mice. Phosphorylation levels of Akt, mTor and p70s6k were increased due to HB-EGF expression in TG mice compared with the NTG mice at 3 and 7 months of age. Additionally, activated Akt, mTor and p70s6k were co-localized with vimentin to cardiac fibroblasts isolated from TG mice. Furthermore, HB-EGF significantly increased phosphorylation levels of Akt, mTor and p70s6k and increased expression of type I collagen in cultured primary cardiac fibroblasts. Rapamycin (Rapa) and CRM197, inhibitors of mTor and HB-EGF respectively, could inhibit the expression of type I collagen in the cultured primary cardiac fibroblasts and Rapa suppressed interstitial fibrosis of the heart tissues in vivo. In addition, a BrdU assay showed that HB-EGF increased proliferation of cardiac fibroblasts by 30% compared with cells without HB-EGF treatment. HB-EGF-induced proliferation was completely diminished in the presence of Rapa. These results suggest that HB-EGF induced heart fibrosis and proliferation of cardiac fibroblasts occurs through activation of the Akt/mTor/p70s6k pathway.

The poor healing capacity and typically incomplete regeneration of injured tendons has made tendon repair as a primacy clinical concern. Several methods for repairing injured tendons have been developed in the last decade. Tendon regeneration using current tissue engineering techniques requires advanced biomaterials to satisfy both microstructural and mechanical criteria. In this study, a novel chitosan (CS)-based scaffold with asymmetric structure was fabricated using a self-deposition technique. The fabricated scaffolds were assessed with regard to the microstructural and mechanical demands of cell ingrowth and the prevention of peritendinous adhesion. In vitro studies showed that rat tendon stem/progenitor cells (TSPCs) seeded onto the CS scaffold displayed higher levels of tenogenic specific genes expression and protein production. Four and six weeks after the implantation of CS scaffolds on full site Achilles tendon defects, in vivo tendon repair was evaluated by histology, immunohistochemistry, immunofluorescence, and mechanical measurements. The production of collagen I (COL1) and collagen III (COL3) demonstrated that the CS scaffolds were capable of inducing conspicuous tenogenic differentiation, higher tenomodulin (TNMD) production, and superior phenotypic maturity, compared with the empty defect group. The introduction of TSPCs into the CS scaffold resulted in a synergistic effect on tendon regeneration and yielded better-aligned collagen fibers with elongated, spindle-shaped cells. These findings indicated that the application of TSPC-seeded CS scaffolds would be a feasible approach for tendon repair. Statement of Significance The poor healing capacity of injured tendons and inevitable peritendinous adhesion has made tendon regeneration a clinical priority. In this study, an asymmetric chitosan scaffold was developed to encapsulate rat tendon stem/progenitor cells (TSPCs), which could induce higher levels of tenogenic specific genes and protein expression. Remarkably, the introduction of TSPCs into the asymmetric chitosan scaffold generated a synergistic effect on in vivo tendon regeneration and lead to better-aligned collagen fibers compared with asymmetric chitosan scaffold alone. This work can provide new guidelines for the structure and property design of cell-seeded scaffolds for tendon regeneration. (C) 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.