Comprehensive biocompatibility testing of a new PMMA-HA bone cement versus conventional PMMA cement in vitro

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Abstract
For more than 50 years PMMA bone cements have been used in orthopaedic surgery. In this study attempts were made to show whether cultured human bone marrow cells (HBMC) show an osteogenetic response resulting in new bone formation, production of extracellular matrix (ECM) and cell differentiation when they were cultured onto polymerized polymethylmethacrylate (PMMA)-hydroxyapatite (HA), conventional PMMA bone cement being taken as reference. Biocompatibility parameters were collagen-I and -II synthesis, the detection of the osteoblast markers alkaline phosphatase (ALP) and osteocalcin, the number of adherent cells and the cytodifferentiation of immunocompetent cells. Cement surface structure, HA stability in culture medium and chemical element analysis of specimens were considered. Fresh marrow cells were obtained from the human femora during hip replacement. Incubation time was up to ten weeks. We used atomic forced microscopy (AFM) and scanning electron microscopy (SEM) for cement specimen analysis. Fluorescent activated cell sorter (FACS), immunohistochemical staining. SEM and light microscopy (LM) served us to judge the cellular morphology. Products of the extracellular matrix were analyzed by protein dot blot analysis, SEM energy dispersive X-ray analysis (SEM-EDX) and Ca2+/PO(4)3- detection. HA particles increased the osteogenetic potential of PMMA bone cement regarding the cellular production of collagen, alkaline phosphatase (AP), the number of osteoblasts and the cellular differentiation pattern in vitro. Both tested cements showed good biocompatibility in a human long-term bone marrow cell-culture system.