Cito-compability analysis of mesenchymal stem cells in platelet rich fibrin matrix (PRFM) for tissue regeneration

Abstract

Mesenchymal stem cells (MSCs) is progenitor cell that capable to differentiate to various mesenchymal or non-mesenchymal lineages. MSC also play a role towards injury during healing and regeneration processes in most tissue. Platelet rich plasma (PRP) had been reported to induce endometrium regeneration, therefore a compacted form of PRP, known as platelet rich fibrin matrix (PRFM), was studied for its capability to host MSC and the behavior of MSC in the matrix. PRP gelation was generated by adding 25 mM of CaCl2 to form a coin shape with diameter of 5 cm. Each matrix was cut into half, seeded directly with bone marrow (hBM-MSC) and umbilical cord MSC (hUC-MSC) with seeding density of 2,000 cell.cm-2. Initial cellular attachment was analyzed using H&E staining, while the cellular viability was assessed quantitatively using MTT assay and qualitatively using Live/Dead staining. hUC-MSC showed the highest viability compared to hBM-MSC when delivered in PRFM, especially during the first and five days incubation. After 10 dan 15 days incubation, the viability had dropped. The Live/Dead staining indicated too crowded population and most cells were dead. This might be due to asphyxia. hUC-MSC and hBM-MSC were seen to proliferated and infiltrated the PRFM over the time, although the ratio of dead cells was more dominant in the later day of incubation. PRFM can be used to deliver MSC to the site injury. However the direct seeding method was not optimal to hold the cells on place during the first 24 h incubation because they were easily detached.