Clinical Ophthalmology (Dec 2023)
Optimizing Platelet and Leucocyte-Rich Plasma as Biomaterials for Ophthalmic Applications: Impact of Centrifugation Speed
Abstract
Alberto Castillo-Macías,1 Judith Zavala,1 Wendy Ortega-Lara,1 Silvia Margarita García-Herrera,2 Jorge E Valdez-García1 1Instituto Tecnológcio y de Estudios Superiores de Monterrey, ITESM, Monterrey, Nuevo León, México; 2Medicina Diagnóstica, Monterrey, Nuevo León, MéxicoCorrespondence: Jorge E Valdez-García, Instituto Tecnológcio y de Estudios Superiores de Monterrey, ITESM, Av. Batallon de San Patricio 112, Real San Agustín, San Pedro Garza García, Monterrey, Nuevo León, 66260, México, Tel +52 81 8888, Email [email protected]: To assess the impact of varying centrifugation speeds on platelet and leucocyte-rich plasma (L-PRP) in liquid and gel form cellularity and growth factor concentrations for potential use against ocular surface disorders.Patients and Methods: L-PRP was collected from 16 healthy subjects using three different centrifugation speeds: 580, 1000, and 2000 g, each for 8 min at 25°C. Platelet and leukocyte counts were automatically evaluated. The concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and transforming growth factor beta 1 (TGF-B1) were measured using enzyme-linked immunosorbent assays. L-PRP gel cellularity was assessed through hematoxylin-eosin and Masson’s trichrome staining, categorized as moderate or abundant, and statistically analyzed. L-PRP gel membrane’s chemical composition was analyzed using Fourier-transform infrared spectroscopy (FTIR), crystallization was investigated with X-ray diffraction (XRD), and ultrastructure was assessed using surface electron microscopy (SEM). Additionally, membrane degradation was evaluated over a 7-day period.Results: No significant differences in cellularity and growth factor concentrations among centrifugation speeds (p > 0.05) were found. Moderate cellularity predominated at 580 g and 2000 g, while abundant cellularity was observed at 1000 g. No significant differences were found techniques (p = 0.16). Masson’s trichrome staining suggested the existence of abundant fibrin at 1000 g but without significant differences (p = 0.07). FTIR analysis exhibited the characteristic fibrin bands at all speeds, and XRD indicated a keratin-like pattern. SEM revealed greater porosity at 580 g and fibrin membrane degradation was lower at this speed (p = 0.0001).Conclusion: Centrifugation speed did not significantly affect growth factor concentration or cellularity in both liquid and gel L-PRP. Further studies should explore the impact of different separation techniques for L-PRP used in ophthalmic applications.Plain Language Summary: Background: Platelet and Leukocyte-rich plasma (L-PRP) is obtained from an individual’s blood. It is currently being utilized successfully and showing encouraging outcomes in ophthalmology for various illnesses. The issue of standardization, particularly the centrifugation speed, is one of the main obstacles to its wider application because the preparation protocols vary depending on the disease type and the setting.Goal: We carried out this study to evaluate the effect of centrifugation speed in obtaining platelet and leucocyte-rich plasma in liquid and gel form and its impact on the quality of the product.Results: We discovered that centrifugation speed affected Platelet and Leukocyte-rich plasma cellularity. Furthermore, the centrifugation speed influenced the platelet and leucocyte-rich plasma gel deterioration after seven days. These findings highlight the significance of standardizing procedures, particularly centrifugation speed, to produce a high-quality product that lasts longer and may yield more encouraging outcomes when applied in the field of ophthalmology.Conclusion: L-PRP is a product with great clinical potential. Standardization is the biggest barrier to widespread use. We observed differences depending on the type of centrifugation speed used for its preparation, which opens the way for us to continue carrying out future studies until we obtain a product with the best therapeutic potential in patients with ophthalmological diseases.Keywords: platelet, leucocytes, centrifugation, growth factors
