Physico-Chemical Factors Influencing Autologous Conditioned Serum Purification

Abstract

Autologous conditioned serum (ACS) is a recent biotherapy based on certain cytokines anti-inflammatory properties mainly used for the reduction of osteoarthritis (OA) symptoms. Here we investigated different physico-chemical factors influencing ACS purification and cytokine production. Human venous blood was incubated in the presence of different diameter beads (respectively 2.5, 3, 3.5, and 4 mm) or glass beads with different types of coating (polished or coated with CrSO4). Sera were recovered, and the concentrations of pro-inflammatory and anti-inflammatory relevant cytokines were measured using Luminex(®) technology. Fresh whole blood incubated for 24 h highly increased production of interleukin (IL)-6 and IL-8 cytokines. At the same time, the concentrations of IL-1β, IL-1 receptor agonist (IL-1Ra), IL-10, and tumor necrosis factor (TNF)-α were slightly induced. The highest cytokine concentrations were obtained with the exposure of whole blood to 3-mm glass beads and 3.5-mm polished beads. The minimum IL-1β/IL-1Ra ratio obtained was 3.2±1.3 after 24-h incubation without any beads. ACS has been shown to alleviate clinical symptoms of OA in clinical studies. This descriptive study demonstrated that different pro- and anti-inflammatory cytokines are present in ACS since no selective anti-inflammatory cytokines were produced based on the different protocols. Furthermore, we showed that CrSO4-treated glass beads are not necessary and that the absence of beads combined with a 24-h incubation could also lead to an enriched serum.