Biofabrication of a novel leukocyte-fibrin-platelet membrane as a cells and growth factors delivery platform for tissue engineering applications

Abstract

Autologous platelet-rich hemocomponents have emerged as potential biologic tools for regenerative purpose, but their therapeutic efficacy still remains controversial. This work represents the characterization study of an innovative autologous leukocyte-fibrin-platelet membrane (LFPm), which we prepared according to a novel protocol involving multiple cycles of apheresis. The high content in fibrinogen gave to our hemocomponent the appearance of a manipulable and suturable membrane with high elasticity and deformation capacity. Moreover, being highly enriched with platelets, leukocytes, and monocytes/macrophages, the LFPm sustained the local release of bioactive molecules (platelet derived growth factor, vascular endothelial growth factor, interleukin-10, and tumour necrosis factor alpha). In parallel, the evaluation of stemness potential highlighted also that the LFPm contained cells expressing pluripotency and multipotency markers both at the messenger ribonucleic acid (NANOG, SOX2, THY1, NT5E, and ENG) and surface-protein level (CD44 /CD73 /CD34 /CD117 /CD31 ). Finally, biodegradation analysis interestingly showed a good stability of the membrane for at least 3 weeks in vitro and 1 week in vivo. In both cases, biodegradation was associated with progressive exposure of fibrin scaffold, loss/migration of cellular elements, and release of growth factors. Overall, collected evidence could shed some light on the regenerative effect that LFPms may exert after the autologous implant on a defect site.

Effects of leukocyte‐ and platelet‐rich fibrin associated or not with bovine bone graft on the healing of bone defects in rats with osteoporosis induced by ovariectomy

“…However, it is important to emphasize that there has never been any published evidence demonstrating that a PRF membrane can maintain spaces for tissue regeneration for sufficient periods of time because of its rapid degradability (Kawase et al, ). Specific in vitro/in vivo analyses of biodegradation profiles of fibrin membranes have been rarely reported in literature (Barbon et al, ). Kawase et al, (), demonstrated that L‐PRF was completely resorbed within 2 weeks after subcutaneous implantation in nude mice.…”

“…Kawase et al, (), demonstrated that L‐PRF was completely resorbed within 2 weeks after subcutaneous implantation in nude mice. Barbon et al, (), produced a innovative autologous leukocyte‐fibrin‐platelet membrane (LFPm), which was prepared according to a novel protocol involving multiple cycles of apheresis. The biodegradation analysis showed a good stability of the membrane for only 1 week in vivo.…”

Platelet-Rich Fibrin Scaffolds for Cartilage and Tendon Regenerative Medicine: From Bench to Bedside

“…The final membrane is highly enriched with platelets, leukocytes, monocytes/macrophages, fibrinogen, and CD34 + cells. Our research group performed a detailed in vitro characterization of the CLP-MB membrane, highlighting its regenerative properties in terms of GF release, mechanical behavior and biodegradation profile [12]. The CLP-MB exhibits good elasticity and high deformation capacity, demonstrating itself to be suitable for implantation in the Knee joint for articular cartilage repair.…”

“…Moreover, this blood-derived membrane is enriched with leukocytes, which play a key role not only in immune and antibacterial responses, but also in the wound healing process [7,8]. Since Choukroun’s PRF was first described, many variations of the original protocol have appeared, resulting in the production of PRF-like materials with different architectures and cell contents [9,10,11,12,13,14,15,16,17]. The fundamental challenge to be overcome remains the concentration of platelets, which should be increased to a minimum of 5 times above baseline values for the hemocomponent to be considered “platelet rich” [18].…”

“…Interestingly, beside the enrichment in platelets and leukocytes, the entrapment of stem-like cells with high regenerative potential within the fibrin network has recently been acknowledged [11,12], providing an even more solid basis for the use of PRF in regenerative medicine.…”