Harmony in Healing: Investigating Platelet-Rich Plasma Activation during Acetylsalicylic Acid Treatment

Abstract

Platelet-rich plasma (PRP) therapy holds promise for treating various clinical conditions. The activation process is crucial in releasing growth factors and cytokines from platelets, enhancing the therapeutic properties of PRP. Standard activation methods involve autologous thrombin or collagen, with variations in efficacy and growth factor release. This study explores the impact of acetylsalicylic acid (ASA), a commonly used antiplatelet drug, on PRP activation. The results indicate that non-activated PRP extracted from the whole blood of ASA-treated patients exhibits increased inflammatory cytokine concentrations, notably TNFa. After activation with autologous thrombin/CaCl₂ or collagen IV, the measured fluorescence intensities suggest varying release patterns between treated and non-treated groups. Understanding the influence of ASA on platelet activation holds implications for personalized medicine and optimizing outcomes for individual patients undergoing PRP therapy. This research sheds light on the potential challenges associated with using antiplatelet drugs, emphasizing the need for careful consideration in tailoring PRP-based regenerative therapies.

Keywords: antiplatelet drugs; collagen; inflammatory cytokines; platelet-rich plasma; regenerative therapies; thrombin.

Figure 1

Platelet-rich plasma collection and activation. PRP was isolated using a whole blood separator (A). PPP (syringe on the left) was collected each time along with the PRP (B) and was used for subsequent thrombin isolation (C). Activation with thrombin resulted in visible clot formation, marked with a black arrow (D). Samples after 60 min activation, from the left: non-activated PRP, PRP activated with thrombin, PRP activated with collagen IV, PRP activated with collagen I, and PPP. PRP—platelet-rich plasma; PPP—platelet-poor plasma.

Figure 2

The comparison between cytokine concentration in non-activated platelet-rich plasma obtained from patients treated with acetylsalicylic acid (ASA) compared to the control group. After activation with autologous thrombin/CaCl_2, the fold increase in fluorescence intensity in the ASA group in comparison to the control group was 1.16 for IL-1A, 1.10 for IL-8, 1.16 for IL-10, 1.06 for MCP-1, 1.11 for INFg, and 1.32 for TNFa (Figure 3). For IL-1B, IL-4, and IL-6, activation with thrombin/CaCl2 resulted in lower fluorescence intensity in the ASA group than in the control. Activation with collagen I increased only the release of MCP-1 (1.15-fold) and TNFa (1.26-fold). In turn, activation with collagen IV resulted in a fold change of 1.61, 1.38, 1.10, 1.22, 1.23, 1.48, and 1.10 for IL-1B, IL-4, IL-6, IL-8, IL-10, IFNg, and TNFa, respectively. *, p-value < 0.05 was considered statistically significant.

Figure 3

The comparison between inflammatory cytokine concentration in platelet-rich plasma activated with calcium chloride (CaCl₂), collagen I (col I), and collagen IV (col IV) obtained from patients treated with acetylsalicylic acid (ASA) compared to the control group.