Abstract
Purpose :
Platelet-rich plasma (PRP) membranes have high potential for use as therapeutic agents in ocular surface disorders. The reported methods vary on centrifugation and PRP separation, which can impact the growth factors concentrations and cellularity, affecting their functionality. We evaluated the impact of the variation in the centrifugation speed on the cellularity and concentration of growth factors of PRP membranes for ophthalmic use.
Methods :
This study was conducted according to the ARVO guidelines for Human research (Ethics Committee 018-2022-CI-R, 2022). PRP was obtained from 16 healthy subjects. The concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF) and transforming growth factor beta 1 (TGF-B1) were evaluated by enzyme-linked immunosorbent assays (ELISA) in PRP obtained with 3 centrifugation speeds: 580 g, 1000 g, and 2000 g (8 minutes at 25 degrees Celsius). Cellularity was assessed by H&E and Masson’s trichrome staining, measured as moderate or abundant, and analyzed by chi-squared test. ANOVA were performed to evaluate the significant difference between the concentrations of growth factors and the centrifugation speed.
Results :
The highest VEGF concentration was 482 pg/ml (SE: 98.65) obtained at 2000 g. The PDGF highest concentration was observed at 1597.75 pg/ml (SE: 60.85) obtained at 580 g. TGF-B1 highest concentration was 843.05 pg/ml (SE: 90.70) obtained at 1000 g. There was no significant difference between the centrifugation speeds and concentration of VEGF, PDGF and TGF-B1 (p>0.05). The observed cellularity had a moderate predominance for the 580 g and 2000 g techniques, while abundant cellularity was observed in the 1000 g technique. No significant difference was found between the evaluated techniques (p=0.16). In the evaluation using Masson's trichrome, precipitates highly suggestive of fibrin were observed abundantly in the 1000 g technique but there was no significant difference between the techniques (p=0.07).
Conclusions :
This study demonstrated that the speed of centrifugation does not impact on the growth factors concentration and cellularity of the PRP. Further studies should be carried out to evaluate the impact of the separation technique to obtain PRP to produce membranes for ophthalmic use.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.