The effect of hyaluronic acid and collagen on growth factors within platelet-rich fibrin: An in-vitro pilot study

Platelet-Rich Fibrin (PRF) is an autologous product enabling physiological platelet activation and fibrin polymerization. It consists of leukocytes, platelets, and a fibrin network enriched with growth factors that accelerate wound healing and promote collagen production. Exploring ways to enhance PRF's regenerative potential and assessing the influence of externally added materials on its growth factors are areas of ongoing research. Hyaluronic acid (HA), a polysaccharide with diverse physicochemical and biological properties, has been widely used as a supportive agent in periodontal therapy. Similarly, collagen, the most abundant protein in the human body, is recognized for its structural support and frequent use in biomedical applications due to its modifiable nature and ability to form cross-links or integrate with biological molecules. Both HA and collagen are likely to impact the mechanical properties of the fibrin matrix in PRF and the growth factors it contains. In this study, venous blood samples were collected from 20 volunteer patients using 7 tubes. The first tube was sent for blood count analysis, while the second tube was used to prepare pure PRF. HA was added to the third tube and collagen to the fourth tube prior to centrifugation. The fifth tube was used to obtain pure PRF for mechanical tests and SEM analysis, whereas the sixth and seventh tubes, containing HA and collagen, respectively, were also used for mechanical and SEM analyses. PRF was obtained by centrifugation at 2700 rpm for 12 minutes, and three experimental groups were established: Control PRF, HA-PRF, and COL-PRF. The growth factors in all PRF samples were quantified using the ELISA method, and PRF membranes underwent tensile and elongation tests with a testometric device. The results showed that the PDGF growth factor was significantly higher in the HA-PRF and COL-PRF groups at 72 hours compared to the control group, though no positive effects were noted for other growth factors. Mechanical testing revealed significantly greater elongation values in the COL-PRF group compared to the control and HA-PRF groups. Similarly, tensile strength was markedly higher in the COL-PRF group than in the control group. These findings suggest that PRF enriched with HA and collagen exhibits improved mechanical properties. The observed enhancements in tensile and elongation strength may positively impact the resorption period of PRF, indicating its potential as an alternative to membranes in bone augmentation procedures.