ABSTRACT
In the cardiac microenvironment, cardiomyocytes (CMs) are embedded in an aligned and structured extracellular matrix (ECM) to maintain the coordinated contractile function of the heart. The cardiac fibroblast (cFB) is the main cell type responsible for producing and remodeling this matrix. In cardiac diseases, however, adverse remodeling and CM death may lead to deterioration of the aligned myocardial structure. Here, we present an in vitro cardiac model system with uniaxial and biaxial constraints to induce (an)isotropy in 3D microtissues, thereby mimicking 'healthy' aligned and 'diseased' disorganized cardiac matrices. A mixture of neonatal mouse CMs and cFBs was resuspended in a collagen-matrigel hydrogel and seeded to form microtissues to recapitulate the in vivo cellular composition. Matrix disarray led to a stellate cell shape and a disorganized sarcomere organization, while CMs in aligned matrices were more elongated and had aligned sarcomeres. Although matrix disarray has no detrimental effect on the force generated by the CMs, it did have a negative effect on the homogeneity of contraction force distribution. Furthermore, proliferation of the cFBs affected microtissue contraction as indicated by the negative correlation between the percentage of cFBs in the microtissues and their beating frequency. These results suggest that in regeneration of the diseased heart, reorganization of the disorganized matrix will contribute to recover the coordinated contraction but restoring the ratio in cellular composition (CMs and cFBs) is also a prerequisite to completely regain tissue function.
Subject(s)
Extracellular Matrix/physiology , Myocardial Contraction/physiology , Myocardium/cytology , Myocytes, Cardiac/physiology , Tissue Engineering/methods , Animals , Animals, Newborn , Anisotropy , Extracellular Matrix/ultrastructure , Finite Element Analysis , Mice , Mice, Inbred C57BL , Microscopy, Confocal , Microscopy, Fluorescence , Myocardium/ultrastructure , Myocytes, Cardiac/cytologyABSTRACT
Carcass properties and meat quality characteristics of 32 Belgian Blue White double-muscled bulls (DM) were compared with those of 59 bulls of the same breed with normal conformation (N). DM showed superior carcass quality as revealed in increased dressing percentage, meat production yield, conformation grade, muscle/fat and muscle/bone ratios (all P < 0·001). Longissimus dorsi (LD) shear force values, drip and cooking losses at 8 days post mortem (pm) were significantly higher (P < 0·001) for DM, whereas sarcomere lengths were not different. Calpain 1 and calpastatin levels at 1 h and 24 h pm were tremendously decreased in DM as were also cathepsin B and L levels at 1 and 8 days pm (P < 0·001). As evident from semi-quantitative SDS-PAGE, these differences were accompanied by higher titin and lower 30 kDa levels (P < 0·001) in DM. Troponin-t levels were not different, but very low. Intramuscular collagen content was significantly lower in DM (P < 0·001). This suggests that lower background toughness in DM was compensated for by reduced post-mortem proteolytic tenderization. Discrepancy with literature reports regarding tenderness of DM might be related to the extreme muscularity of the Belgian Blue White breed, compared to other DM breeds. The results also suggest that reduced protein turnover might be involved in the muscle hypertrophy phenomenon within this breed, because of likely reduced levels of calpains and cathepsins in living DM animals.
