Optimized Autologous Serum and Activated Leukocyte-Poor Platelet-Rich Plasma Eye Drops Production Protocol: Effect of Fasting,Filtering,and Complement Inactivation on the Concentration of Growth Factors and Lipids
Restricted accessResearch articleFirst published online January, 2026
Optimized Autologous Serum and Activated Leukocyte-Poor Platelet-Rich Plasma Eye Drops Production Protocol: Effect of Fasting,Filtering,and Complement Inactivation on the Concentration of Growth Factors and Lipids
To analyze the effect of fasting on volume, growth factor, and lipid concentration in eye drops derived from autologous serum (AS) and leukocyte-poor platelet-rich plasma (LP-PRP). To assess changes in protein and growth factor concentration after filtration with three different syringe filters and after complement inactivation with heat.
Methods:
Blood samples from 10 volunteers after a fasting and nonfasting period were harvested on two separate visits. The total volume gained of AS and LP-PRP was measured. Also, the concentration of lipids, albumin, lysozyme, beta-nerve growth factor (β-NGF), transforming growth factor-β1 (TGF-β1), epidermal growth factor (EGF), hepatocyte growth factor (HGF), platelet-derived growth factor-BB (PDGF-BB), and vascular endothelial growth factor-A (VEGF-A) were quantified in each preparation. The effect of microfiltration was measured in pooled AS and LP-PRP samples before and after filtration with polyethersulfone, polyvinylidene fluoride, or mixed cellulose esters (MCE) membrane syringe filters. The extent of complement inactivation was quantified after incubation at +56°C for 15–240 min.
Results:
The total volume of AS and LP-PRP and the protein and growth factor concentration remained similar after fasting. Increased triacyl glycerides and similar phospholipid levels were observed in nonfasting samples. Except for a minimal decrease in TGF-β1 after filtration with an MCE membrane filter, filtering did not reduce the concentration of growth factors in AS and LP-PRP. Heat inactivation considerably reduced levels of lysozyme, EGF, HGF, and β-NGF in AS and LP-PRP.
Conclusions:
AS should be produced after a fasting period to facilitate the filtration. However, nonfasting does not affect concentrations of relevant biologically active constituents and phospholipids.
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