Abstract: Introduction: Orthobiologics such as Platelet-Rich Plasma (PRP) and Injectable Platelet-Rich Fibrin (i-PRF) have emerged as promising tools in regenerative medicine. However, the lack of methodological standardization and the still limited comparative characterization between these products represent significant barriers to their optimized clinical application. This comparative laboratory study aimed to characterize and differentiate PRP and i-PRF, focusing on their cellular composition, obtained volume, and concentration of Platelet-Derived Growth Factor (PDGF-BB). Materials and Methods: This study was conducted with 34 healthy individuals. Peripheral blood samples were collected from all participants. PRP was obtained using a modified double-spin centrifugation protocol, whereas i-PRF was prepared using a modified low-speed centrifugation technique. Cellularity (platelet and leukocyte counts), final produced volume, and PDGF-BB concentration were assessed using complete blood count analysis and enzyme-linked immunosorbent assay (ELISA), respectively. Statistical analysis was performed using Generalized Linear Models (GLM). Results: Both protocols resulted in significant increases in platelet and leukocyte concentrations compared to baselines values. PRP showed significantly higher platelet and leukocyte concentrations compared with i-PRF, as well as markedly higher PDGF-BB levels. In contrast, i-PRF yielded a substantially greater final volume and enabled a higher absolute delivery of total leukocytes, whereas PRP delivered a greater absolute number of platelets. Female Sex, presence of comorbidities, and increased abdominal circumference positively influenced product volume and cellular composition. Discussion: Although both PRP and i-PRF effectively concentrate blood-derived components, they present distinct biological profiles regarding cellularity, volume, and growth factor availability. These findings indicate that PRP and i-PRF are not interchangeable, and their clinical use should be guided by the target tissue, therapeutic mechanism, and evidence from randomized clinical trials.