Interaction of platelet-rich concentrate with bone graft materials: an in vitro study.

Objective: Platelet-rich concentrate (PRC) is in routine use for orthopaedic and maxilofacial surgery and is frequently combined with bone graft materials to fill bony defects and enhance healing. Numerous studies have been performed investigating the efficacy of PRC to enhance bone healing in which a variety of graft materials have been combined with varying degrees of success. Here, we sought to determine the effect of combining PRC with different graft materials on human bone marrow stromal cell (hBMSC) proliferation, osteoblastic differentiation, and bone formation. Methods: Our central hypothesis is that PRC is not a true osteogenic agent but rather is osteopromotive, with cell fate determination being dependent on additional signals derived from the microenvironment. Experiments were performed with low passage (maximum 3) hBMSCs that were maintained in the presence of ascorbic acid-2-phosphate and beta-glycerol phosphate. Dexamethasone was excluded from these studies. PRC and graft materials were retained within well inserts and clotted by addition of bovine thrombin. Cell proliferation was determined by DNA content, osteoblastic commitment, and differentiation by alkaline phosphatase activity and matrix mineralization. Results: Combining PRC with the graft materials increased proliferation above that seen with the graft materials alone; however, only demineralized bone matrix (DBM) and allograft were capable of increasing proliferation above that seen with PRC alone. The increased proliferation observed in the presence of PRC coincided with decreased normalized alkaline phosphatase activity, suggesting decreased osteoblastic differentiation. However, at later time points, PRC increased mineralization compared with DBM, collagen, or beta tricalcium phosphate alone. When compared with PRC alone, addition of DBM or allograft decreased mineralization. Collagen gave rise to a small increase in mineralization, whereas beta tricalcium phosphate yielded the same level of mineralization as PRC alone. Conclusions: The data obtained from these in vitro investigations demonstrate that the cellular responses induced by PRC and bone graft materials in hBMSC can be significantly (positively or negatively) modified by adding the agents in combination. These in vitro data highlight the need to consider the potential interaction between biologic agents when added in combination.