RESUMEN
BACKGROUND: Blood platelets (PLTs) link the processes of hemostasis and inflammation. Recent studies have demonstrated that PLTs promote immunity and inflammation mainly by means of the CD40/CD40L pathway. Our objective was to describe the accumulation of cytokines in PLT concentrates during storage. STUDY DESIGN AND METHODS: Pools of PLT concentrates were prepared, separated from plasma, and resuspended in clinical-grade storage medium; samples were taken on Days 0, 1, 2, 3, and 5 for analysis, without replacement (i.e., without soluble protein dilution). Interleukin (IL)-6, IL-8, PLT-derived growth factor (PDGF)-AA, soluble CD40 ligand (sCD40L), RANTES, and transforming growth factor-beta production were measured by specific enzyme-linked immunosorbent assays. RESULTS: Over time, the levels of RANTES, IL-8, and IL-6 were stable. In contrast, the levels of PDGF-AA and sCD40L increased. Ex vivo production of sCD40L was quantified at levels sufficient to induce B-cell effects based on previous studies of in vitro induced B-cell activation and differentiation by sCD40L. Cytokine and/or chemokine levels were generally higher in PLT concentrate supernatants and/or PLT lysates in comparison to PLT-free plasma, allowing the determination of which cytokine and/or chemokine was absorbed or secreted by transfusion-grade PLTs over time. CONCLUSION: Our data provide evidence that stored PLTs contain molecules with known immunomodulatory competence and secrete them differentially over time during storage for transfusion purposes.
Asunto(s)
Plaquetas/inmunología , Conservación de la Sangre , Factores Inmunológicos/metabolismo , Plaquetas/metabolismo , Ligando de CD40/análisis , Quimiocina CCL5/análisis , Citaféresis , Citocinas/análisis , Humanos , Interleucinas/análisis , Transfusión de Plaquetas/métodos , Transfusión de Plaquetas/normas , Factor de Crecimiento Derivado de Plaquetas/análisis , Factores de TiempoRESUMEN
This in vivo study investigated the efficiency of an injectable calcium phosphate bone substitute (IBS) for bone regenerative procedures through non-destructive three-dimensional (3D) micro-tomographic (microCT) imaging, biomechanical testing with a non-destructive micro-indentation technique and 2D scanning electron microscopy (SEM) analysis. The injectable biomaterial was obtained by mixing a biphasic calcium phosphate (BCP) ceramic mineral phase and a cellulosic polymer. The BCP particles were 200-500 microm or 80-200 microm in diameter. The injectable material was implanted for 6 weeks into critical-sized bone defects at the distal end of rabbit femurs. Extensive new bone apposition was noted with both 2D and 3D techniques. Micro-CT showed that newly formed bone was in perfect continuity with the trabecular host bone structure and demonstrated the high interconnectivity of the restored bone network. For both IBS formulations, SEM and microCT gave very close measurements. The only detected significant difference concerned the amount of newly formed bone obtained with IBS 80-200 that appeared significantly higher with microCT analysis than with SEM (p=0.00007). Student t-tests did not show any significant difference in the amount of newly formed bone and remaining ceramic obtained from microCT analysis or SEM. Regression analysis showed satisfactory correlation between both the amount of newly formed bone and remaining ceramic obtained from microCT or SEM. For IBS 200-500, the newly formed bone rate inside the defect was 28.0+/-5.2% with SEM and yield strength of the samples was 18.8+/-5.4 MPa. For IBS 80-200, the newly formed bone rate inside the defect was 31.7+/-5.1% with SEM and yield strength of the samples was 26.8+/-4.5 MPa. Yield strength appeared well correlated with the amount of newly formed bone, specially observed with microCT. This study showed the ability of non-destructive techniques to investigate biological and mechanical aspects of bone replacement with injectable biomaterials.