Sample Collection and Measurements
Blood samples were obtained from patients with diabetic foot ulcers
before treatment was initiated. A total of 5 mL blood samples were taken
from the volunteers: 3 mL blood samples were placed in tubes containing
anticoagulants to obtain whole blood, and 2 mL blood samples were placed
into dry tubes. Serum samples were obtained by centrifuging the dry tube
blood samples at 3500 g for 10 minutes. Whole blood and serum samples
were kept at -80oC until the study day. The body mass
index (BMI) was calculated by dividing the body weight in kilograms by
the square of the body height in meters (kg/m2).
In the whole blood samples, the amount of glycated haemoglobin (HbA1c)
was determined on an analyzer (Arkray Adams HA-8160 from Japan) working
with a high pressure liquid chromatography method. The serum levels of
low-density lipoprotein (LDL), high-density lipoprotein (HDL),
triglyceride, total cholesterol and glucose were measured by
autoanalysis using photometric methods (Architect C 16200 Abbott
Laboratories Inc., Abbott Park, IL, USA). The levels of CRP were
detected by a BNR II SYSTEM SN 442176 instrument.
The serum IMA levels in both groups were measured according to the
method described by Bar-Or et al.19 A 200 µL serum
sample was placed in a glass tube, 50 µL cobalt chloride added and the
mixture then incubated for 10 min at 24oC before 50 µL
dithiothreitol was added to the mixture. After 2 minutes, 1 mL of sodium
chloride solution was added. The IMA concentrations were calculated by
measuring the absorbance of the coloured complex at 470 nm wavelength.
The serum IMA levels were expressed as absorubans (ABSU).