Influence of calcium chloride and aprotinin in the in vivo biological performance of a composite combining biphasic calcium phosphate granules and fibrin sealant.
ABSTRACT Highly bioactive biomaterials have been developed to replace bone grafts in orthopedic revision and maxillofacial surgery for bone augmentation. A mouldable, self-hardening material can be obtained by combining TricOs Biphasic Calcium Phosphate Granules and Tissucol Fibrin Sealant. Two components, calcium chloride and antifibrinolytic agents (aprotinin), are essential for the stability of the fibrin clot. The ingrowth of cells in composites combining sealants without calcium chloride or with a low concentration of aprotinin was evaluated in vivo in an experiment on rabbits. Bone colonization was compared using TricOs alone or with the composite made from TricOs and the standard fibrin sealant. Without the addition of calcium chloride, the calcium ions released by the ceramic component interacted with the components of the sealant too late to stabilize the clot. With a low concentration of aprotinin, the degradation of the clot occurred more quickly, leading to the absence of a scaffold on which the bone cells could colonize the composite. Our results indicate that a stable fibrin scaffold is crucial for bone colonization. The low calcium chloride and low aprotinin groups have shown lower bone growth. Further studies will be necessary to determine the minimal amount of antifibrinolytic agent (aprotinin) necessary to allow the same level of osteogenic activity as the TricOs-fibrin glue composite.
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ABSTRACT: Membranes of the autologous blood-derived biomaterial platelet-rich fibrin (PRF) were functionalized by incorporation of alkaline phosphatase (ALP), an enzyme involved in mineralization of bone, and subsequently incubated in calcium glycerophosphate (CaGP) solution to induce PRFs mineralization with calcium phosphate (CaP) to improve PRFs suitability as a material for bone replacement. Incorporated ALP retained its bioactivity and induced formation of CaP material within PRF membranes, as confirmed by SEM, EDS, FTIR, and von Kossa staining. The mass percentage attributable to CaP was quantified by lyophilization and measurement of the remaining mass fraction as well as by TGA. Cytocompatibility tests (LDH, MTT, and WST) with SAOS-2 cells showed that mineralized PRF did not release substances detrimental to cell vitality. Live/dead staining and SEM showed that mineralized PRF was colonized by cells. The results show that hydrogel biomaterials such as PRF can be mineralized through functionalization with ALP.Journal of Biomedical Materials Research Part A 02/2012; 100(5):1335-46. · 2.83 Impact Factor
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ABSTRACT: The limited intrinsic repair capacity of articular cartilage has led to the investigation of different treatment options to promote its regeneration. The delivery of hydrogels containing stem or progenitor cells and growth factor releasing microspheres represents an attractive approach to cartilage repair. In this study, the influence of the encapsulating hydrogel on the ability of progenitor cells coupled with TGF-β3 releasing microspheres to form cartilaginous tissue was investigated. Fibrin, agarose and gellan gum hydrogels containing TGF-β3 loaded gelatin microspheres and progenitor cells derived from the infrapatellar fat-pad of the knee were cultured for 21 days in a chemically defined media. In the presence of TGF-β3 releasing microspheres, gellan gum hydrogels were observed to facilitate greater cell proliferation than fibrin or agarose hydrogels. Histological and biochemical analysis of the hydrogels indicated that fibrin was the least chondro-inductive of the three hydrogels, while agarose and gellan gum appeared to support more robust cartilage formation as demonstrated by greater sGAG accumulation within these constructs. Gellan gum hydrogels also stained more intensely for collagen type II and collagen type I, suggesting that although total collagen synthesis was higher in these constructs, that the phenotype may be more fibrocartilaginous in nature than normal hyaline cartilage. This study demonstrates how the encapsulating hydrogel can have a significant impact on the ability of stem cells to form cartilage when incorporated into a growth factor delivery system.Biomedical Materials 03/2013; 8(3):035004. · 2.17 Impact Factor
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ABSTRACT: Calcium phosphate cements (CPCs) and fibrin glue (FG) are used for surgical applications. Their combination is promising to create bone substitutes able to promote cell attachment and bone remodeling. This study proposes a novel approach to create CPC-FG composites by simultaneous CPC setting and FG fibrinogenesis. CPC-FG composites were obtained by mixing CPC powders, i.e. α-tricalcium phosphate, dicalcium phosphate anhydrous and precipitated hydroxyapatite, with FG powder components, i.e. fibrinogen and thrombin, and a 2% Na(2) HPO(4) solution. To study the effect of FG quantity and fibrinogenesis kinetics, long and fast setting FGs were evaluated in amounts of 0.125, 0.250, and 0.500 mL on CPC-FG composites. Physicochemical, interconnectivity, and mechanical properties were measured. Scanning electron microscopy, Micro-computed tomography (μ-CT), X-ray diffraction, and Fourier transform Infrared spectroscopy (FTIR) analyzed morphology, structure, crystallographic, and chemical composition, respectively. FG fibrinogenesis was performed within the CPC. FTIR confirmed this and its interfacial bonding with CPC. μ-CT confirmed a good FG distribution. FG addition affected the CPC when compared with pristine CPC. Adding FG to CPC changed their morphology, density, porosity, setting, cohesion, injectability, interconnectivity, crystallographic and chemical composition and mechanical properties. Moreover, 0.500 mL of long setting FG modified the observed fracture behavior of the CPC-FG. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.Journal of Biomedical Materials Research Part A 08/2012; · 2.83 Impact Factor