| 1. |
- Wu, Lisha, 1984, et al.
(author)
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Initial DNA Interactions of the Binuclear Threading Intercalator Λ,Λ-[μ-bidppz(bipy)4Ru2]4+: An NMR Study with [d(CGCGAATTCGCG)]2
- 2013
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In: Chemistry - A European Journal. - : Wiley. - 0947-6539 .- 1521-3765. ; 19:17, s. 5401-5410
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Journal article (peer-reviewed)abstract
- Binuclear polypyridine ruthenium compounds have been shown to slowly intercalate into DNA, following a fast initial binding on the DNA surface. For these compounds, intercalation requires threading of a bulky substituent, containing one RuII, through the DNA base-pair stack, and the accompanying DNA duplex distortions are much more severe than with intercalation of mononuclear compounds. Structural understanding of the process of intercalation may greatly gain from a characterisation of the initial interactions between binuclear RuII compounds and DNA. We report a structural NMR study on the binuclear RuII intercalator -B (-[-bidppz-(bipy)4Ru2]4+; bidppz=11,11’-bis(di-pyrido[3,2-:2’,3’-c]phenazinyl, bipy = 2,2’-bipyridine) mixed with the palindromic DNA [d(CGCGAATTCGCG)]2. Threading of -B depends on the presence and length of AT stretches in the DNA. Therefore, the latter was selected to promote initial binding, but due to the short stretch of AT base pairs, final intercalation is prevented. Structural calculations provide a model for the interaction: -B is trapped in a well-defined surface-bound state consisting of an eccentric minor-groove binding. Most of the interaction enthalpy originates from electrostatic and van der Waals contacts, whereas intermolecular hydrogen bonds may help to define a unique position of -B. Molecular dynamics simulations show that this minor-groove binding mode is stable on a nanosecond scale. To the best of our knowledge, this is the first structural study by NMR spectroscopy on a binuclear Ru compound bound to DNA. In the calculated structure, one of the positively charged Ru2+ moieties is near the central AATT region; this is favourable in view of potential intercalation as observed by optical methods for DNA with longer AT stretches. Circular dichroism (CD) spectroscopy suggests that a similar binding geometry is formed in mixtures of -B with natural calf thymus DNA. The present minor-groove binding mode is proposed to represent the initial surface interactions of binuclear RuII compounds prior to intercalation into AT-rich DNA.
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| 2. |
- Barba, Albert, et al.
(author)
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Impact of Biomimicry in the Design of Osteoinductive Bone Substitutes : Nanoscale Matters
- 2019
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In: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 11:9, s. 8818-8830
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Journal article (peer-reviewed)abstract
- Bone apatite consists of carbonated calcium-deficient hydroxyapatite (CDHA) nanocrystals. Biomimetic routes allow fabricating synthetic bone grafts that mimic biological apatite. In this work, we explored the role of two distinctive features of biomimetic apatites, namely, nanocrystal morphology (plate vs needle-like crystals) and carbonate content, on the bone regeneration potential of CDHA scaffolds in an in vivo canine model. Both ectopic bone formation and scaffold degradation were drastically affected by the nanocrystal morphology after intramuscular implantation. Fine-CDHA foams with needle-like nanocrystals, comparable in size to bone mineral, showed a markedly higher osteoinductive potential and a superior degradation than chemically identical coarse-CDHA foams with larger plate-shaped crystals. These findings correlated well with the superior bone-healing capacity showed by the fine-CDHA scaffolds when implanted intraosseously. Moreover, carbonate doping of CDHA, which resulted in small plate-shaped nanocrystals, accelerated both the intrinsic osteoinduction and the bone healing capacity, and significantly increased the cell-mediated resorption. These results suggest that tuning the chemical composition and the nanostructural features may allow the material to enter the physiological bone remodeling cycle, promoting a tight synchronization between scaffold degradation and bone formation.
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| 3. |
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| 4. |
- Barba, Albert, et al.
(author)
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Osteogenesis by foamed and 3D-printed nanostructured calcium phosphate scaffolds: Effect of pore architecture
- 2018
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In: Acta Biomaterialia. - : Elsevier BV. - 1742-7061 .- 1878-7568. ; 79, s. 135-147
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Journal article (peer-reviewed)abstract
- There is an urgent need of synthetic bone grafts with enhanced osteogenic capacity. This can be achieved by combining biomaterials with exogenous growth factors, which however can have numerous undesired side effects, but also by tuning the intrinsic biomaterial properties. In a previous study, we showed the synergistic effect of nanostructure and pore architecture of biomimetic calcium deficient hydroxyapatite (CDHA) scaffolds in enhancing osteoinduction, i.e. fostering the differentiation of mesenchymal stem cells to bone forming cells. This was demonstrated by assessing bone formation after implanting the scaffolds intramuscularly. The present study goes one step forward, since it analyzes the effect of the geometrical features of the same CDHA scaffolds, obtained either by 3D-printing or by foaming, on the osteogenic potential and resorption behaviour in a bony environment. After 6 and 12 weeks of intraosseous implantation, both bone formation and material degradation had been drastically affected by the macropore architecture of the scaffolds. Whereas nanostructured CDHA was shown to be highly osteoconductive both in the robocast and foamed scaffolds, a superior osteogenic capacity was observed in the foamed scaffolds, which was associated with their higher intrinsic osteoinductive potential. Moreover, they showed a significantly higher cell-mediated degradation than the robocast constructs, with a simultaneous and progressive replacement of the scaffold by new bone. In conclusion, these results demonstrate that the control of macropore architecture is a crucial parameter in the design of synthetic bone grafts, which allows fostering both material degradation and new bone formation. Statement of Significance 3D-printing technologies open new perspectives for the design of patient-specific bone grafts, since they allow customizing the external shape together with the internal architecture of implants. In this respect, it is important to design the appropriate pore geometry to maximize the bone healing capacity of these implants. The present study analyses the effect of pore architecture of nanostructured hydroxyapatite scaffolds, obtained either by 3D-printing or foaming, on the osteogenic potential and scaffold resorption in an in vivo model. While nanostructured hydroxyapatite showed excellent osteoconductive properties irrespective of pore geometry, we demonstrated that the spherical, concave macropores of foamed scaffolds significantly promoted both material resorption and bone regeneration compared to the 3D-printed scaffolds with orthogonal-patterned struts and therefore prismatic, convex macropores.
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| 5. |
- Barba, Albert, et al.
(author)
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Osteoinduction by Foamed and 3D-Printed Calcium Phosphate Scaffolds : Effect of Nanostructure and Pore Architecture
- 2017
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In: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 9:48, s. 41722-41736
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Journal article (peer-reviewed)abstract
- Some biomaterials are osteoinductive, that is, they are able to trigger the osteogenic process by inducing the differentiation of mesenchymal stem cells to the osteogenic lineage. Although the underlying mechanism is still unclear, microporosity and specific surface area (SSA) have been identified as critical factors in material-associated osteoinduction. However, only sintered ceramics, which have a limited range of porosities and SSA, have been analyzed so far. In this work, we were able to extend these ranges to the nanoscale, through the foaming and 3D-printing of biomimetic calcium phosphates, thereby obtaining scaffolds with controlled micro- and nanoporosity and with tailored macropore architectures. Calcium-deficient hydroxyapatite (CDHA) scaffolds were evaluated after 6 and 12 weeks in an ectopic-implantation canine model and compared with two sintered ceramics, biphasic calcium phosphate and β-tricalcium phosphate. Only foams with spherical, concave macropores and not 3Dprinted scaffolds with convex, prismatic macropores induced significant ectopic bone formation. Among them, biomimetic nanostructured CDHA produced the highest incidence of ectopic bone and accelerated bone formation when compared with conventional microstructured sintered calcium phosphates with the same macropore architecture. Moreover, they exhibited different bone formation patterns; in CDHA foams, the new ectopic bone progressively replaced the scaffold, whereas in sintered biphasic calcium phosphate scaffolds, bone was deposited on the surface of the material, progressively filling the pore space. In conclusion, this study demonstrates that the high reactivity of nanostructured biomimetic CDHA combined with a spherical, concave macroporosity allows the pushing of the osteoinduction potential beyond the limits of microstructured calcium phosphate ceramics.
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| 6. |
- Braesch-Andersen, Anna, et al.
(author)
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Application of phase-field fracture theories and digital volume correlation to synchrotron X-ray monitored fractures in human trabecular bone : A case study
- 2022
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In: Journal of The Mechanical Behavior of Biomedical Materials. - : Elsevier. - 1751-6161 .- 1878-0180. ; 135
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Journal article (peer-reviewed)abstract
- Fracture processes of trabecular bone have been studied using various approaches over the years. However, reliable methods to analyse fracture at the single trabecula level are limited. In this study, a digital volume correlation (DVC) and a phase-field fracture model are applied and contrasted for human trabecular bone to analyse its failure under global compression at high resolution.A human trabecular bone sample was fractured in situ under synchrotron-based X-ray micro computed to-mography (CT). Reconstructed CT data was then used in DVC algorithms to obtain high-resolution displacement fields in the bone at different load steps. A high-resolution specimen-specific structural mesh was discretized from the CT data and used for the phase-field simulation of the fracturing bone.The DVC analysis showed opening mode cracks as well as shear mode cracks. Strains in cracked regions were analysed. The load distribution in the trabecular structure resulted in two completely separated fracture regions in the sample body. A phenomenon that was also captured in the phase-field model. The results encourage us to believe improvements in boundary conditions and material models are worthwhile pursuing. Findings in this study support further development of a phase-field method to analyse fracture in samples with complex morphology, such as trabecular bone, and the capacity of DVC to quantify strains and slowly growing stable fractures during step-wise loading of trabecular bone.
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| 7. |
- Diez-Escudero, Anna, et al.
(author)
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Effect of calcium phosphate heparinization on the in vitro inflammatory response and osteoclastogenesis of human blood precursor cells
- 2019
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In: Journal of Tissue Engineering and Regenerative Medicine. - : WILEY. - 1932-6254 .- 1932-7005. ; 13:7, s. 1217-1229
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Journal article (peer-reviewed)abstract
- The immobilization of natural molecules on synthetic bone grafts stands as a strategy to enhance their biological interactions. During the early stages of healing, immune cells and osteoclasts (OC) modulate the inflammatory response and resorb the biomaterial, respectively. In this study, heparin, a naturally occurring molecule in the bone extracellular matrix, was covalently immobilized on biomimetic calcium-deficient hydroxyapatite (CDHA). The effect of heparin-functionalized CDHA on inflammation and osteoclastogenesis was investigated using primary human cells and compared with pristine CDHA and beta-tricalcium phosphate (beta-TCP). Biomimetic substrates led to lower oxidative stresses by neutrophils and monocytes than sintered beta-TCP, even though no further reduction was induced by the presence of heparin. In contrast, heparinized CDHA fostered osteoclastogenesis. Optical images of stained TRAP positive cells showed an earlier and higher presence of multinucleated cells, compatible with OC at 14 days, while pristine CDHA and beta-TCP present OC at 21-28 days. Although no statistically significant differences were found in the OC activity, microscopy images evidenced early stages of degradation on heparinized CDHA, compatible with osteoclastic resorption. Overall, the results suggest that the functionalization with heparin fostered the formation and activity of OC, thus offering a promising strategy to integrate biomaterials in the bone remodelling cycle by increasing their OC-mediated resorption.
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| 8. |
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| 9. |
- Díez-Escudero, Anna, et al.
(author)
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Heparinization of Beta Tricalcium Phosphate: Osteo-immunomodulatory Effects
- 2018
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In: Advanced Healthcare Materials. - : Wiley-VCH Verlagsgesellschaft. - 2192-2640 .- 2192-2659. ; 7:5
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Journal article (peer-reviewed)abstract
- Immune cells play a vital role in regulating bone dynamics. This has boosted the interest in developing biomaterials that can modulate both the immune and skeletal systems. In this study, calcium phosphates discs (i.e., beta-tricalcium phosphate, β-TCP) are functionalized with heparin to investigate the effects on immune and stem cell responses. The results show that the functionalized surfaces downregulate the release of hydrogen peroxide and proinflammatory cytokines (tumor necrosis factor alpha and interleukin 1 beta) from human monocytes and neutrophils, compared to nonfunctionalized discs. The macrophages show both elongated and round shapes on the two ceramic substrates, but the morphology of cells on heparinized β-TCP tends toward a higher elongation after 72 h. The heparinized substrates support rat mesenchymal stem cell (MSC) adhesion and proliferation, and anticipate the differentiation toward the osteoblastic lineage as compared to β-TCP and control. The coupling between the inflammatory response and osteogenesis is assessed by culturing MSCs with the macrophage supernatants. The downregulation of inflammation in contact with the heparinized substrates induces higher expression of bone-related markers by MSCs.
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| 10. |
- Diez-Escudero, Anna, et al.
(author)
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Hexagonal pore geometry and the presence of hydroxyapatite enhance deposition of mineralized bone matrix on additively manufactured polylactic acid scaffolds
- 2021
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In: Materials science & engineering. C, biomimetic materials, sensors and systems. - : Elsevier. - 0928-4931 .- 1873-0191. ; 125
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Journal article (peer-reviewed)abstract
- Additive manufacturing (AM) has revolutionized the design of regenerative scaffolds for orthopaedic applications, enabling customizable geometric designs and material compositions that mimic bone. However, the available evidence is contradictory with respect to which geometric designs and material compositions are optimal. There is a lack of studies that systematically compare different pore sizes and geometries in conjunction with the presence or absence of calcium phosphates. We therefore evaluated the physicochemical and biological properties of additively manufactured scaffolds based on polylactic acid (PLA) in combination with hydroxyapatite (HA). HA was either incorporated in the polymeric matrix or introduced as a coating, yielding 15 and 2% wt., respectively. Pore sizes of the scaffolds varied between 200 and 450 um and were shaped either triangularly or hexagonally. All scaffolds supported the adhesion, proliferation and differentiation of both primary mouse osteoblasts and osteosarcoma cells up to four weeks, with only small differences in the production of alkaline phosphatase (ALP) between cells grown on different pore geometries and material compositions. However, mineralization of the PLA scaffolds was substantially enhanced in the presence of HA, either embedded in the PLA matrix or as a coating at the surface level, and by larger hexagonal pores. In conclusion, customized HA/PLA composite porous scaffolds intended for the repair of critical size bone defects were obtained by a cost-effective AM method. Our findings indicate that the analysis of osteoblast adhesion and differentiation on experimental scaffolds alone is inconclusive without the assessment of mineralization, and the effects of geometry and composition on bone matrix deposition must be carefully considered in order to understand the regenerative potential of experimental scaffolds.
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