Determination of Endogenous Cytokines in

This study objectively characterized the microenvironment of indolent, chronic wounds by developing a method to extract and identify minute quantities of cytokines. Utilizing recombinant DNA technology, the research aimed to establish values for endogenous cytokine levels before initiating treatment with exogenous agents. The methodology involved the application of porous, inert hydrophilic dextranomer beads, which were tested for their absorption capacities using in vitro experiments with albumin, and subsequently applied to chronic human pressure ulcers. Protein levels were quantitatively assessed through the Bradford protein assay, while enzyme-linked immunosorbent assay (ELISA) techniques were employed to measure the levels of specific growth factors, including platelet-derived growth factor (PDGF)-AB, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and transforming growth factor-Beta (TGF-B). Results indicated a recovery rate of 88% to 98% of known albumin amounts and over 90% for laced cytokines. However, in the 20 grade III/IV pressure ulcers studied, while protein concentrations were consistent, the endogenous cytokine levels exhibited substantial variability. Notably, PDGF-AB levels ranged from 49 to 867 pg/mL, bFGF from 47 to 697 pg/mL, and EGF from undetectable to 247.5 pg/mL, whereas TGF-B was absent in 17 samples. This technique is considered valuable for measuring cytokine levels, revealing much lower concentrations in chronic compared to acute wounds.