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Critical Limb Ischaemia (CLI) is
defined as the presence of rest pain, ulcers and/or gangrene in a limb for over
2 weeks. Associated exercise intolerance is caused by muscle fibre atrophy,
fibro- fatty infiltration, nerve dysfunction, mitochondrial damage and
myofibril disorder. We aimed to determine the behaviour of satellite cells,
responsible for the repair and regeneration of damaged muscle, in repairing the
damage caused to critically ischaemic adult human skeletal muscle. CD34, pax7
and MyoD are all markers of satellite cells at different stages of the cell
cycle and allow for an assessment of their number and activity in ischaemia.
Local ethical committee approval and informed consent was obtained. Samples of
harvested gastrocnemius muscle of patients undergoing major perigenicular
amputation for CLI (n = 10) were analysed and compared to a control group
undergoing coronary artery bypass grafting (n = 10). Using
immunohistochemistry, the expression of pax7, CD34 and MyoD was assessed in
five sequential blinded randomly generated fields. Statistical testing of the
data collected was made via the Mann Whitney U test. Protein electrophoresis
was used to confirm overall protein expression of the satellite cell markers.
There was a significant increase in the number of satellite cells observed in
CLI muscle sections as demonstrated by the expression of pax7 (2.4×？fold p < 0.0001). CD34 expressing？Haematopoietic Stem Cells？(HSCs) and satellite cells were also
more abundant, with a 2×？fold
increase observed (p < 0.0001) whilst those cells expressing both CD34 and
pax7 and identified as quiescent satellite cells, were significantly greater in
number in the CLI samples (2.9×？fold
p < 0.0001), confirmed via immunohistochemistry and protein electrophoresis.
There was a significant decrease in the expression of MyoD positive or
activated satellite cells (p < 0.0001). This indicates an increase in the
proliferation of the satellite cell population as a response to CLI but less
active cells are observed.