Juan Manuel Fernández

Development of an osteoconductive PCL-PDIPF-hydroxyapatite composite scaffold for bone tissue engineering.

Hydroxyapatite (HAP)‐containing poly‐ε‐caprolactone (PCL)–polydiisopropyl fumarate (PDIPF) composite (Blend) was developed as an alternative for bone tissue engineering. The physicochemical, mechanical and biocompatibility properties of these composites were evaluated using two osteoblast‐like cell lines (UMR106 and MC3T3E1) and compared with the blend without HAP and PCL/HAP films. The increment in the elastic modulus and the decrease in the elongation‐at‐break of Blend–HAP suggest that the mechanical properties of the HAP scaffolds have improved significantly. The addition of HAP to both PCL and Blend significantly improves the cell biocompatibility and osteogenicity of the scaffolds. Evidence for this notion is based in several observations: (a) HAP–polymer increases proliferation of osteoblastic cells; (b) HAP included in the blend increases the ALP expression in UMR106 cells; (c) HAP–Blend increases the type‐I collagen production in both cell lines, and d) higher levels of the osteogenic transcription factor Runx‐2 were detected when MC3T3E1 osteoblasts were induced to differentiate and mineralize on HAP–polymer scaffolds. In conclusion, a novel biocompatible HAP–Blend composite with uniform dispersion of semi‐nano HAP particles and good interphase compatibility has been prepared successfully. The development of HAP–Blend composite, with improved physical, mechanical and osteoinductive properties, may potentially be used in bone tissue‐engineering applications. Copyright

Characterization of Poly(ε-caprolactone)/Polyfumarate Blends as Scaffolds for Bone Tissue Engineering

There is considerable interest in the design of polymeric biomaterials that can be used for the repair of bone defects. In this study, we used ultrasound to prepare a compatibilized blend of poly(ε-caprolactone) (PCL) and poly(diisopropyl fumarate) (PDIPF). The formation of post-sonication inter-polymer coupling products was verified by SEC analysis of a blend with azo-labeled PDIPF. We also analyzed the physicochemical and mechanical properties of the compatibilized blend. When compared to PCL alone, the PCL/PDIPF blend showed no difference in its resistance as evaluated by the elastic modulus, although it did show a 50% decrease in ultimate tensile stress (P < 0.05) and an 84% decrease in elongation-at-break (P < 0.05). However, the mechanical properties of this blend were comparable to those of trabecular bone. We next evaluated biocompatibility of the PCL/PDIPF blend, and of homo-polymeric PCL and PDIPF films for comparison, with UMR106 and MC3T3E1 osteoblastic cells. Osteoblasts plated on the compatibilized blend adhered and proliferated more than on either homo-polymer, showed a greater number of cellular processes with a better organized actin cytoskeleton and expressed more type-I collagen and mineral, both markers of osteoblast phenotype. These results support the hypothesis that this new compatibilized blend could be useful in future applications for bone regeneration.

Strontium ranelate prevents the deleterious action of advanced glycation endproducts on osteoblastic cells via calcium channel activation

Accumulation of advanced glycation endproducts (AGEs) in bone tissue occurs in ageing and in Diabetes mellitus, and is partly responsible for the increased risk of low-stress bone fractures observed in these conditions. In this study we evaluated whether the anti-osteoporotic agent strontium ranelate can prevent the deleterious effects of AGEs on bone cells, and possible mechanisms of action involved. Using mouse MC3T3E1 osteoblastic cells in culture we evaluated the effects of 0.1mM strontium ranelate and/or 100 μg/ml AGEs-modified bovine serum albumin (AGEs-BSA) on cell proliferation, osteogenic differentiation and pro-inflammatory cytokine production. We found that AGEs-BSA alone decreased osteoblastic proliferation and differentiation (P<0.01) while increasing IL-1β and TNFα production (P<0.01). On its own, strontium ranelate induced opposite effects: an increase in osteoblast proliferation and differentiation (P<0.01) and a decrease in cytokine secretion (P<0.01). Additionally, strontium ranelate prevented the inhibitory and pro-inflammatory actions of AGEs-BSA on osteoblastic cells (P<0.01). These effects of strontium ranelate were blocked by co-incubation with either the MAPK inhibitor PD98059, or the calcium channel blocker nifedipine. We also evaluated by Western blotting the activation status of ERK (a MAPK) and b-catenin. Activation of both signaling pathways was decreased by AGEs treatment, and this inhibitory effect was prevented if AGEs were co-incubated with strontium ranelate (P<0.01). On its own, strontium ranelate increased both pERK and activated b-catenin levels. In conclusion, this study demonstrates that strontium ranelate can prevent the deleterious in vitro actions of AGEs on osteoblastic cells in culture by mechanisms that involve calcium channel, MAPK and b-catenin activation.