Contreras-Raggio, J. I., Arancibia, C. T., Millan, C., Ploeg, H. L., Aiyangar, A., & Vivanco, J. F. (2022). Height-to-Diameter Ratio and Porosity Strongly Influence Bulk Compressive Mechanical Properties of 3D-Printed Polymer Scaffolds. Polymers, 14(22), 5017.
Abstract: Although the architectural design parameters of 3D-printed polymer-based scaffolds-porosity, height-to-diameter (H/D) ratio and pore size-are significant determinants of their mechanical integrity, their impact has not been explicitly discussed when reporting bulk mechanical properties. Controlled architectures were designed by systematically varying porosity (30-75%, H/D ratio (0.5-2.0) and pore size (0.25-1.0 mm) and fabricated using fused filament fabrication technique. The influence of the three parameters on compressive mechanical properties-apparent elastic modulus E-app, bulk yield stress sigma(y) and yield strain epsilon(y)-were investigated through a multiple linear regression analysis. H/D ratio and porosity exhibited strong influence on the mechanical behavior, resulting in variations in mean E-app of 60% and 95%, respectively. sigma(y) was comparatively less sensitive to H/D ratio over the range investigated in this study, with 15% variation in mean values. In contrast, porosity resulted in almost 100% variation in mean sigma(y) values. Pore size was not a significant factor for mechanical behavior, although it is a critical factor in the biological behavior of the scaffolds. Quantifying the influence of porosity, H/D ratio and pore size on bench-top tested bulk mechanical properties can help optimize the development of bone scaffolds from a biomechanical perspective.
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Millan, C., Vivanco, J. F., Benjumeda-Wijnhoven, I. M., Bjelica, S., & Santibanez, J. F. (2018). Mesenchymal Stem Cells and Calcium Phosphate Bioceramics: Implications in Periodontal Bone Regeneration. Adv.Exp.Med.Biol., 1107, 91–112.
Abstract: In orthopedic medicine, a feasible reconstruction of bone structures remains one of the main challenges both for healthcare and for improvement of patients' quality of life. There is a growing interest in mesenchymal stem cells (MSCs) medical application, due to their multilineage differentiation potential, and tissue engineering integration to improve bone repair and regeneration. In this review we will describe the main characteristics of MSCs, such as osteogenesis, immunomodulation and antibacterial properties, key parameters to consider during bone repair strategies. Moreover, we describe the properties of calciumphosphate (CaP) bioceramics, which demonstrate to be useful tools in combination with MSCs, due to their biocompatibility, osseointegration and osteoconduction for bone repair and regeneration. Also, we overview the main characteristics of dental cavity MSCs, which are promising candidates, in combination with CaP bioceramics, for bone regeneration and tissue engineering. The understanding of MSCs biology and their interaction with CaP bioceramics and other biomaterials is critical for orthopedic surgical bone replacement, reconstruction and regeneration, which is an integrative and dynamic medical, scientific and bioengineering field of research and biotechnology.
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Selcho, M., Millan, C., Palacios-Munoz, A., Ruf, F., Ubillo, L., Chen, J. T., et al. (2017). Central and peripheral clocks are coupled by a neuropeptide pathway in Drosophila. Nat. Commun., 8, 13 pp.
Abstract: Animal circadian clocks consist of central and peripheral pacemakers, which are coordinated to produce daily rhythms in physiology and behaviour. Despite its importance for optimal performance and health, the mechanism of clock coordination is poorly understood. Here we dissect the pathway through which the circadian clock of Drosophila imposes daily rhythmicity to the pattern of adult emergence. Rhythmicity depends on the coupling between the brain clock and a peripheral clock in the prothoracic gland (PG), which produces the steroid hormone, ecdysone. Time information from the central clock is transmitted via the neuropeptide, sNPF, to non-clock neurons that produce the neuropeptide, PTTH. These secretory neurons then forward time information to the PG clock. We also show that the central clock exerts a dominant role on the peripheral clock. This use of two coupled clocks could serve as a paradigm to understand how daily steroid hormone rhythms are generated in animals.
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Sundram, V., Ng, F. S., Roberts, M. A., Millan, C., Ewer, J., & Jackson, F. R. (2012). Cellular Requirements for LARK in the Drosophila Circadian System. J. Biol. Rhythms, 27(3), 183–195.
Abstract: RNA-binding proteins mediate posttranscriptional functions in the circadian systems of multiple species. A conserved RNA recognition motif (RRM) protein encoded by the lark gene is postulated to serve circadian output and molecular oscillator functions in Drosophila and mammals, respectively. In no species, however, has LARK been eliminated, in vivo, to determine the consequences for circadian timing. The present study utilized RNA interference (RNAi) techniques in Drosophila to decrease LARK levels in clock neurons and other cell types in order to evaluate the circadian functions of the protein. Knockdown of LARK in timeless (TIM)- or pigment dispersing factor (PDF)-containing clock cells caused a significant number of flies to exhibit arrhythmic locomotor activity, demonstrating a requirement for the protein in pacemaker cells. There was no obvious effect on PER protein cycling in lark interference (RNAi) flies, but a knockdown within the PDF neurons was associated with increased PDF immunoreactivity at the dorsal termini of the small ventral lateral neuronal (s-LNv) projections, suggesting an effect on neuropeptide release. The expression of lark RNAi in multiple neurosecretory cell populations demonstrated that LARK is required within pacemaker and nonpacemaker cells for the manifestation of normal locomotor activity rhythms. Interestingly, decreased LARK function in the prothoracic gland (PG), a peripheral organ containing a clock required for the circadian control of eclosion, was associated with weak population eclosion rhythms or arrhythmicity.
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Uranga, R. M., Millan, C., Barahona, M. J., Recabal, A., Salgado, M., Martinez, F., et al. (2017). Adenovirus-mediated suppression of hypothalamic glucokinase affects feeding behavior. Sci Rep, 7, 13 pp.
Abstract: Glucokinase (GK), the hexokinase involved in glucosensing in pancreatic beta-cells, is also expressed in arcuate nucleus (AN) neurons and hypothalamic tanycytes, the cells that surround the basal third ventricle (3V). Several lines of evidence suggest that tanycytes may be involved in the regulation of energy homeostasis. Tanycytes have extended cell processes that contact the feeding-regulating neurons in the AN, particularly, agouti-related protein (AgRP), neuropeptide Y (NPY), cocaine-and amphetamine-regulated transcript (CART) and proopiomelanocortin (POMC) neurons. In this study, we developed an adenovirus expressing GK shRNA to inhibit GK expression in vivo. When injected into the 3V of rats, this adenovirus preferentially transduced tanycytes. qRT-PCR and Western blot assays confirmed GK mRNA and protein levels were lower in GK knockdown animals compared to the controls. In response to an intracerebroventricular glucose injection, the mRNA levels of anorexigenic POMC and CART and orexigenic AgRP and NPY neuropeptides were altered in GK knockdown animals. Similarly, food intake, meal duration, frequency of eating events and the cumulative eating time were increased, whereas the intervals between meals were decreased in GK knockdown rats, suggesting a decrease in satiety. Thus, GK expression in the ventricular cells appears to play an important role in feeding behavior.
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Wijnhoven, I. B., Vallejos, R., Santibanez, J. F., Millan, C., & Vivanco, J. F. (2020). Analysis of cell-biomaterial interaction through cellular bridge formation in the interface between hGMSCs and CaP bioceramics. Sci Rep, 10(1), 14 pp.
Abstract: The combination of biomaterials and stem cells for clinical applications constitute a great challenge in bone tissue engineering. Hence, cellular networks derived from cells-biomaterials crosstalk have a profound influence on cell behaviour and communication, preceding proliferation and differentiation. The purpose of this study was to investigate in vitro cellular networks derived from human gingival mesenchymal stem cells (hGMSCs) and calcium phosphate (CaP) bioceramic interaction. Biological performance of CaP bioceramic and hGMSCs interaction was evaluated through cell adhesion and distribution, cellular proliferation, and potential osteogenic differentiation, at three different times: 5 h, 1 week and 4 weeks. Results confirmed that hGMSCs met the required MSCs criteria while displaying osteogenic differentiaton capacities. We found a significant increase of cellular numbers and proliferation levels. Also, protein and mRNA OPN expression were upregulated in cells cultured with CaP bioceramic by day 21, suggesting an osteoinductible effect of the CaP bioceramic on hGMSCs. Remarkably, CaP bioceramic aggregations were obtained through hGMSCs bridges, suggesting the in vitro potential of macrostructures formation. We conclude that hGMSCs and CaP bioceramics with micro and macropores support hGMSC adhesion, proliferation and osteogenic differentiation. Our results suggest that investigations focused on the interface cells-biomaterials are essential for bone tissue regenerative therapies.
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