Meat Science and Muscle Biology Symposium: manipulating meat tenderness by increasing the turnover of intramuscular connective tissue.

Controlled reduction of the connective tissue contribution to cooked meat toughness is an objective that would have considerable financial impact in terms of added product value. The amount of intramuscular connective tissue in a muscle appears connected to its in vivo function, so reduction of the overall connective tissue content is not thought to be a viable target. However, manipulation of the state of maturity of the collagenous component is a biologically viable target; by increasing connective tissue turnover, less mature structures can be produced that are functional in vivo but more easily broken down on cooking at temperatures above 60°C, thus improving cooked meat tenderness. Recent work using cell culture models of fibroblasts derived from muscle and myoblasts has identified a range of factors that alter the activity of the principal enzymes responsible for connective tissue turnover, the matrix metalloproteinases (MMP). Fibroblasts cultured from 3 different skeletal muscles from the same animal show different cell proliferation and MMP activity, which may relate to the different connective tissue content and architecture in functionally different muscles. Expression of MMP by fibroblasts is increased by vitamins that can counter the negative effects of oxidative stress on new collagen synthesis. Preliminary work using in situ zymography of myotubes in culture also indicates increased MMP activity in the presence of epinephrine and reactive oxidative species. Comparison of the relative changes in MMP expression from muscle cells vs. fibroblasts shows that myoblasts are more responsive to a range of stimuli. Muscle cells are likely to produce more of the total MMP in muscle tissue as a whole, and the expression of latent forms of the enzymes (i.e., pro-MMP) may vary between oxidative and glycolytic muscle fibers within the same muscle. The implication is that the different muscle fiber composition of different muscles eaten as meat may influence the potential for manipulation of their connective tissue turnover.

Phenotypic differences in matrix metalloproteinase 2 activity between fibroblasts from 3 bovine muscles.

Different muscles in a beef carcass are known to respond differently to the same stimulus or animal growth pattern or both. This may complicate the search by the meat industry for production methods to render meat tender. One of the major differences between muscles in the same carcass is in the expression of intramuscular connective tissue. Current study investigates the existence of a phenotypic difference among fibroblasts from 3 bovine skeletal muscles as exemplified by the expression of matrix metalloproteinases (MMP) the main enzymes responsible for connective tissue turnover. The sensitivity of phenotypic differences to cell culture conditions (passage number, presence of growth factors from fetal serum) was also examined. Fibroblasts, the main cells responsible for the production and turnover of collagen were isolated from LM, semitendinosus (STN), and sternomandibularis (SMD) muscles from a bull calf and grown in DMEM, 10% fetal bovine serum, and 5% CO(2). Cell doubling times, survival time, resting expression, and activity of MMP and the effect of serum withdrawal in the culture media on MMP expression and activity were determined for each cell line during 15 passages. Fibroblasts isolated from the 3 muscles had different growth potentials. The shortest (P < 0.0001) cell doubling times for almost every passage were found in cells from STN muscle. Cells from the LM had a shorter (P < 0.0001) survival time in comparison with STN and SMD. Cells derived from the STN had greater values (P > 0.05) of MMP-2 activity in comparison with LM and SMD cells until passage 4. At passage 15, no activity was detected for any cell line. Serum withdrawal generally reduced MMP-2 activation but did not eliminate differences in activity between fibroblasts from the 3 muscles. These results suggest that fibroblasts from different locations are phenotypically different and may respond differently to the same growth or nutritional stimulus in vitro. This may be related to in vivo differences in accumulation, maturity, and turnover of collagen, and ultimately meat tenderness. These findings may be important for selecting a management strategy for improving meat tenderness by manipulation of animal growth; a strategy applied to the whole animal may work for some muscles but not for others.

Disparity of dietary effects on collagen characteristics and toughness between two beef muscles.

Manipulation of growth rate and/or diet has been shown to affect protein turnover and may be used to improve beef quality. This trial was conducted to evaluate the effects of average daily gain (ADG) and diet on animal performance, collagen characteristics and beef quality of two different muscles; longissimus dorsi (LD) and semitendinosus (ST). Seventy six Hereford and Angus steers were assigned to three dietary management regimens for finishing: high grain diet based on corn (n=28), alfalfa pasture (n=22) and grass pasture (n=26). Average daily gains were greater (P<0.001) in Herefords vs. Angus and for corn- vs. pasture finished cattle. Overall, total collagen content was greater (P<0.001) and the percentage of total collagen that was heat soluble was lower (P<0.0001) for ST than for LD muscle. The lowest (P<0.05) values for both total and heat soluble collagens were found in animals finished on corn. WBSF values for LD were greater (P<0.01) in grass-fed vs. alfalfa- and grain-finished cattle while there was no difference in WBSF values for ST between grass- and corn-fed animals. No correlation between ADG and WBSF was observed for any muscle. ADG was not correlated with collagen solubility in ST, but was correlated (P<0.05) with collagen solubility in LD. A key finding is that growth rate affected heat soluble collagen in the two muscles to a different extent. In conclusion, this study shows that different feeding strategies may not influence the tenderness of all muscles in a similar way.