Abstract

Platelet-derived growth factor-enriched plasma (PD-GFEP) remains an essential biological supplement in regenerative medicine. Like other platelet derivatives, the criteria for producing high-quality products for clinical use depend on the specific application. This highlights the importance of exploring suitable activation stimuli and effective methods for production and preservation to maintain functionality for biological approvals or allogeneic use. This study aimed to produce a lyophilized platelet-derived growth factor-enriched plasma (L-PD-GFEP) meeting high-quality standards for clinical application and to evaluate its in vitro functionality after reconstitution. Optimal parameters for producing high-quality platelet-rich plasma (PRP) suitable for clinical use were identified through a meta-analysis. The collection of leukocyte-poor, volume-reduced PRP (LP-PRP) was optimized using a three-step centrifugation process, resulting in undetectable white and red blood cell counts, a volume reduction of 78.72% ± 2.2, a concentration (X value) of 4.09 ± 0.4, and a capture efficiency of 86.32% ± 5.7. Activation of LP-PRP with thrombin, CaCl2, or cold incubation (4°C) was evaluated using 96-well plate-based aggregometry, which showed distinct absorbance patterns over time. The in vitro functional assessment indicated that lyophilized PD-GFEP prepared by cold incubation contains more active factors that promote fibroblast proliferation and peripheral blood mononuclear cell activation than those prepared with thrombin and CaCl2. These findings provide current information for standardizing the criteria for producing and lyophilizing PD-GFEP and demonstrate its in vitro functionality after reconstitution.

Key words: Activation, Functionality, Growth factors, Lyophilization, Platelets