Activated macrophages for treating skin ulceration: gene expression in human monocytes after hypo-osmotic shock

Abstract

SUMMARY Macrophages play a major role in almost all stages of the complex process of wound healing. It has been previously shown that the incorporation of a hypo‐osmotic shock step, in the process of monocyte‐concentrate preparation from a blood unit, induces monocyte/macrophage activation. As the macrophages are produced using a unique, closed and sterile system, they are suitable for local application on ulcers in elderly and paraplegic patients. Enhanced phagocytosis by the activated cells, as well as increased secretion of cytokines such as IL‐1, IL‐6, were detected in a recent study which are in accord with the very encouraging clinical results. In the present study, we used DNA microarrays to analyse the differential gene expressions of the hypo‐osmotic shock‐activated monocytes/macrophages and compare them to non‐treated cells. Of the genes that exhibited differences of expression in the activated cell population, 94% (68/72) displayed increased activity. The mRNA levels of 43/68 of these genes (63%) were found to be 1·5‐fold or higher (1·5–7·98) in the activated macrophages cell population as compared to the non‐treated cells. Only four genes were found to have lower mRNA levels in the activated cells, with ratios of expression of 0·62–0·8, which may suggest that the changes are insignificant. A significant number of the genes that showed increased levels of expression is known to be directly involved in macrophage function and wound healing. This may correlate with the increased secretion of different cytokines by the activated macrophages depicted previously. Other groups of genes expressed are known to be involved in important pathways such as neuronal growth and function, developmental defects and cancer. The hypo‐osmotic shock induces a gene expression profile of cytokines and receptors in the activated cells. These may evoke potential abilities to produce a variety of protein products needed in the wound healing process and may bring to light possibilities for other therapeutic applications of these cells.

Techniques for Wound Healing with a Focus on Pressure Ulcers Elimination

“…However, prior to ulcer application the monocytes / macrophages must first be activated. This can be accomplished by application of hypoosmotic shock during the process of monocyte-concentrate preparation from a blood unit [133].…”

Ex Vivo Activated Human Macrophages Improve Healing, Remodeling, and Function of the Infarcted Heart

“…Purity of cells was assessed by flow cytometry-activated cell sorter (FACS). 15 Monocyte activation was confirmed by enzyme-linked immunosorbent assay (ELISA) kits for interleukin levels in the supernatants obtained from the activated and nonactivated monocyte cultures. 14 Phagocytosis capacity of fluorescent latex beads, by activated and nonactivated monocytes collected from the culture bags, was evaluated by FACS.…”

Enhancement of an Allogeneic GM-CSF-Secreting Breast Cancer Vaccine by Immunomodulatory Doses of Cyclophosphamide and Doxorubicin

“…Immune tolerance represents the second major barrier to the effectiveness of cancer vaccines (reviewed in ref 29). Unlike infectious challenges, tumor cells arise endogenously.…”