High-Speed Video Microscopy for Primary Ciliary Dyskinesia Diagnosis: A Study of Ciliary Motility Variations with Time and Temperature

Primary ciliary dyskinesia (PCD) is a rare disease resulting from a defect in ciliary function that generates, among other issues, chronic upper and lower respiratory tract infections. European guidelines recommend studying ciliary function (pattern (CBP) and frequency (CBF)), together with characteristic clinical symptoms, as one of the definitive tests. However, there is no “gold standard”. The present study aims to use high-speed video microscopy to describe how CBF and CBP alter over time and at different temperatures to reduce the error rate in the diagnosis of PCD. Samples of nasal epithelium from 27 healthy volunteers were studied to assess CBF and CBP at 0, 3, 24, 48, and 72 h, at room temperature and 4 °C. It was observed that CBF increased while CBP became dyskinetic, both at room temperature and at 4 °C, as time passed, especially after 3 h. In order to preserve all ciliary function parameters and to perform a reliable analysis to improve the diagnostic process of PCD, analysis should be performed within the first 3 h of sample collection, preferably in reference centers

A standardised approach to the biomechanical evaluation of tracheal grafts

[EN] The ideal tracheal substitute must have biomechanical properties comparable to the native trachea, but currently there is no standardised approach to evaluating these properties. Here we propose a novel method for evaluating and comparing the properties of tracheal substitutes, thus systematising both measurement and data curation. This system was tested by comparing native rabbit tracheas to frozen and decellularised specimens and determining the histological characteristics of those specimens. We performed radial compression tests on the anteroposterior tracheal axis and longitudinal axial tensile tests with the specimens anastomosed to the jaw connected to a measuring system. All calculations and results were adjusted according to tracheal size, always using variables relative to the tracheal dimensions, thus permitting comparison of different sized organs. The biomechanical properties of the decellularised specimens were only slightly reduced compared to controls and significant in regard to the maximum stress withstood in the longitudinal axis (-0.246 MPa CI [-0.248, -0.145] MPa) and the energy stored per volume unit (-0.124 mJ & BULL;mm(-3) CI [-0.195, -0.055] mJ & BULL;mm(-3)). The proposed method is suitable for the systematic characterisation of the biomechanical properties of different tracheal substitutes, regardless of the size or nature of the substitute, thus allowing for direct comparisons.This research was funded by the 2018 Spanish Society of Thoracic Surgery grant to National Multicentric Study [Number 180101 to N.J.M.-H.] and [PI16-01315 to M.M.-R.] from the Instituto de Salud Carlos III. CIBERER is funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and the Instituto de Salud Carlos III, with assistance from the European Regional Development Fund.Martínez-Hernández, NJ.; Más Estellés, J.; Milián-Medina, L.; Martínez-Ramos, C.; Cerón-Navarro, J.; Galbis-Caravajal, J.; Roig-Bataller, A.... (2021). A standardised approach to the biomechanical evaluation of tracheal grafts. Biomolecules. 11(10):1-12. https://doi.org/10.3390/biom11101461S112111

Propuesta de lugares de interés geológico asociados a bloques y cordones de origen tsunamítico en la costa de Menorca (Baleares)

La costa SE de Menorca es el tramo donde han impactado los tsunamis procedentes de Argelia, dejando como testimonio de su impacto bloques aislados y cordones de bloques de dimensión métrica, con unas características sedimentarias propias de flujos de tsunami. La morfología, morfometría, orientación e imbricación de estos campos de bloques constituyen un rasgo geomorfológico característico del impacto de tsunami en la costa rocosa de las islas Baleares. Por su elevado interés científico se proponen dos emplazamientos como Lugar de Interés Geológico en la isla de Menorca, ya que presentan elevado interés científico, turístico y didáctico como ejemplos representativos de bloques de tsunamis en las costas rocosas del mediterráneo occidental

Optimization of a decellularized protocol of porcine tracheas. Long-term effects of cryopreservation. A histological study

[EN] Objective: The aim of this study was to optimize a decellularization protocol in the trachea of Sus scrofa domestica (pig) as well as to study the effects of long-term cryopreservation on the extracellular matrix of decellularized tracheas. Methods: Porcine tracheas were decellularized using Triton X-100, SDC, and SDS alone or in combination. The effect of these detergents on the extracellular matrix characteristics of decellularized porcine tracheas was evaluated at the histological, biomechanical, and biocompatibility level. Morphometric approaches were used to estimate the effect of detergents on the collagen and elastic fibers content as well as on the removal of chondrocytes from decellularized organs. Moreover, the long-term structural, ultrastructural, and biomechanical effect of cryopreservation of decellularized tracheas were also estimated. Results: Two percent SDS was the most effective detergent tested concerning cell removal and preservation of the histological and biomechanical properties of the tracheal wall. However, long-term cryopreservation had no an appreciable effect on the structure, ultrastructure, and biomechanics of decellularized tracheal rings. Conclusion: The results presented here reinforce the use of SDS as a valuable decellularizing agent for porcine tracheas. Furthermore, a cryogenic preservation protocol is described, which has minimal impact on the histological and biomechanical properties of decellularized porcine tracheas.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by grants MAT2016-76039-C4-2-R (MST) and PID2019-106099RB-C42 (MM) from the Ministry of Economy and Competitiveness of the Spanish Government, by grant PI16-01315 from the ISCIII (Ministerio de Ciencia, Innovacion y Universidades, Spain), and by grant PROMETEO/2020/069 (CC) from the local government of the Comunitat Valenciana (Spain), CIBER-BBN and CIBERER are funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and the Instituto de Salud Carlos III, with assistance from the European Regional Development Fund.Milián, L.; Sancho-Tello, M.; Roig-Soriano, J.; Foschini, G.; Martínez-Hernández, NJ.; Más Estellés, J.; Ruiz-Sauri, A.... (2021). Optimization of a decellularized protocol of porcine tracheas. Long-term effects of cryopreservation. A histological study. The International Journal of Artificial Organs. 44(12):998-1012. https://doi.org/10.1177/03913988211008912S9981012441

Angiotensin II Induces Neutrophil Accumulation In Vivo Through Generation and Release of CXC Chemokines

Background— Angiotensin II (Ang II) is implicated in the development of cardiac ischemic disorders in which prominent neutrophil accumulation occurs. Ang II can be generated intravascularly by the renin-angiotensin system or extravascularly by mast cell chymase. In this study, we characterized the ability of Ang II to induce neutrophil accumulation. Methods and Results— Intraperitoneal administration of Ang II (1 nmol/L) induced significant neutrophil recruitment within 4 hours (13.3±2.3x106 neutrophils per rat versus 0.7±0.5x106 in control animals), which disappeared by 24 hours. Maximal levels of CXC chemokines were detected 1 hour after Ang II injection (577±224 pmol/L cytokine-inducible neutrophil chemoattractant [CINC]/keratinocyte-derived chemokine [KC] versus 5±3, and 281±120 pmol/L macrophage inflammatory protein [MIP-2] versus 14±6). Intravital microscopy within the rat mesenteric microcirculation showed that the short-term (30 to 60 minutes) leukocyte–endothelial cell interactions induced by Ang II were attenuated by an anti-rat CINC/KC antibody and nearly abolished by the CXCR2 antagonist SB-517785-M. In human umbilical vein endothelial cells (HUVECs) or human pulmonary artery media in culture, Ang II induced interleukin (IL)-8 mRNA expression at 1, 4, and 24 hours and the release of IL-8 at 4 hours through interaction with Ang II type 1 receptors. When HUVECs were pretreated with IL-1 for 24 hours to promote IL-8 storage in Weibel-Palade bodies, the Ang II–induced IL-8 release was more rapid and of greater magnitude. Conclusions— Ang II provokes rapid neutrophil recruitment, mediated through the release of CXC chemokines such as CINC/KC and MIP-2 in rats and IL-8 in humans, and may contribute to the infiltration of neutrophils observed in acute myocardial infarction.Mata Roig, Manuel, [email protected] ; Cortijo Gimeno, Julio, [email protected] ; Morcillo Sanchez, Esteban Jesus, [email protected] ; Jose, Peter J., [email protected] ; Sanz Ferrando, Maria Jesus, [email protected]

Correlation between biological responses in vitro and in vivo to Ca-doped sol-gel coatings assessed using proteomic analysis

Poor correlation between the results of in vitro testing and the subsequent in vivo experiments hinders the design of biomaterials. Thus, new characterisation methods are needed. This study used proteomic and histological techniques to analyse the effects of Ca-doped biomaterials in vitro and in vivo and verify the correlation between the two systems. The sol-gel route was employed to synthesise coatings functionalised with 0.5 and 5 wt% of CaCl2. Morphology of the coatings was examined using SEM; the Ca2+ ion release from the materials was analysed by means of ICP-AES spectroscopy. The osteogenic and inflammatory responses were inspected in vitro in human osteoblasts (HOb) and TPH-1 monocytes. The in vivo experiments used a rabbit model. The nLC-MS/MS-based proteomic methods were utilised to analyse the proteins adhering to the material samples incubated with human serum or examine protein expression in the tissues close to the implants. Ca-doped biomaterials caused a remarkable increase in the adsorption of coagulation-related proteins, both in vitro (PLMN, THRB, FIBA and VTNC) and in vivo (FBLN1, G1U978). Enhanced affinity to these materials was also observed for proteins involved in inflammation (CO5, C4BPA, IGHM and KV302 in vitro; CARD6, DDOST and CD14 in vivo) and osteogenic functions (TETN, PEDF in vitro; FBN1, AHSG, MYOC in vivo). The results obtained using different techniques were well matched, with a good correlation between the in vitro and in vivo experiments. Thus, the proteomic analysis of biological responses to biomaterials in vitro is a useful tool for predicting their impact in vivo

Micropartículas de PLLA y CHT como andamiaje para la regeneración del cartílago articular : modelo animal.

The avascular nature of cartilaginous tissue has historically lead to bad prognosis in osteochondral injuries. One of the possible treatment options of these injuries is the use of scaffolds, being superior to other options that obtain a fibrous cartilage as a result. We have elaborated PLLA and CHT microspheres as a scaffold for the treatment of osteochondral injuries carried out in albine New Zealand rabbits. They were distributed into groups with different proportions of microspheres, having also a control group with untreated injuries. Native cartilage of the contralateral knees was also analysed. Samples were evaluated in order to establish the quality of the cartilage obtained (using de macroscopic ICRS, microscopic ICRS II scales and a histomorphometric study).The groups with microspheres obtained a regeneration cartilage with hyaline characteristics, a good cell distribution and regular surface. The control group resulted in a cartilage with worse organization and an irregular surface

La histología en tiempos de pandemia y más allá

Conferencia virtual brindada en el marco del I Congreso de Educación en Ciencias Biológicas, llevado a cabo los días 27 y 28 de noviembre de 2020 en la Facultad de Odontología de la Universidad Nacional de La Plata.Para acceder al video y para su mejor visualización, hacer clic en "Enlace externo".Facultad de Odontologí

Novel Tissue-Engineered Multimodular Hyaluronic Acid-Polylactic Acid Conduits for the Regeneration of Sciatic Nerve Defect

The gold standard for the treatment of peripheral nerve injuries, the autograft, presents several drawbacks, and engineered constructs are currently suitable only for short gaps or small diameter nerves. Here, we study a novel tissue-engineered multimodular nerve guidance conduit for the treatment of large nerve damages based in a polylactic acid (PLA) microfibrillar structure inserted inside several co-linear hyaluronic acid (HA) conduits. The highly aligned PLA microfibers provide a topographical cue that guides axonal growth, and the HA conduits play the role of an epineurium and retain the pre-seeded auxiliary cells. The multimodular design increases the flexibility of the device. Its performance for the regeneration of a critical-size (15 mm) rabbit sciatic nerve defect was studied and, after six months, very good nerve regeneration was observed. The multimodular approach contributed to a better vascularization through the micrometrical gaps between HA conduits, and the pre-seeded Schwann cells increased axonal growth. Six months after surgery, a cross-sectional available area occupied by myelinated nerve fibers above 65% at the central and distal portions was obtained when the multimodular device with pre-seeded Schwann cells was employed. The results validate the multi-module approach for the regeneration of large nerve defects and open new possibilities for surgical solutions in this field.This research was funded by the Spanish Government’s State Research Agency (AEI) through projects DPI2015-72863-EXP and RTI2018-095872-B-C22/ERDF. Funding for open access charge: Universitat Politècnica de València