scholarly journals Real-Time Wireless Platform for In Vivo Monitoring of Bone Regeneration

Sensors ◽  
2020 ◽  
Vol 20 (16) ◽  
pp. 4591 ◽  
Author(s):  
Pablo Blázquez-Carmona ◽  
Manuel Sanchez-Raya ◽  
Juan Mora-Macías ◽  
Juan Antonio Gómez-Galán ◽  
Jaime Domínguez ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Real Time ◽  
Ex Vivo ◽  
Low Cost ◽  
Bone Lengthening ◽  
Bone Callus ◽  
Reliable Solution ◽  
Regeneration Processes

For the monitoring of bone regeneration processes, the instrumentation of the fixation is an increasingly common technique to indirectly measure the evolution of bone formation instead of ex vivo measurements or traditional in vivo techniques, such as X-ray or visual review. A versatile instrumented external fixator capable of adapting to multiple bone regeneration processes was designed, as well as a wireless acquisition system for the data collection. The design and implementation of the overall architecture of such a system is described in this work, including the hardware, firmware, and mechanical components. The measurements are conditioned and subsequently sent to a PC via wireless communication to be in vivo displayed and analyzed using a developed real-time monitoring application. Moreover, a model for the in vivo estimation of the bone callus stiffness from collected data was defined. This model was validated in vitro using elastic springs, reporting promising results with respect to previous equipment, with average errors and uncertainties below 6.7% and 14.04%. The devices were also validated in vivo performing a bone lengthening treatment on a sheep metatarsus. The resulting system allowed the in vivo mechanical characterization of the bone callus during experimentation, providing a low-cost, simple, and highly reliable solution.

scholarly journals Modeling Immune Checkpoint Inhibitor Efficacy in Syngeneic Mouse Tumors in an Ex Vivo Immuno-Oncology Dynamic Environment

2020 ◽  
Vol 21 (18) ◽  
pp. 6478
Author(s):  
Daniel T. Doty ◽  
Julia Schueler ◽  
Vienna L. Mott ◽  
Cassie M. Bryan ◽  
Nathan F. Moore ◽  
...  
Keyword(s):  
Real Time ◽  
Mouse Models ◽  
Immune Checkpoint ◽  
Checkpoint Inhibitors ◽  
Ex Vivo ◽  
Confocal Imaging ◽  
Checkpoint Blockade ◽  
Syngeneic Mouse

The immune checkpoint blockade represents a revolution in cancer therapy, with the potential to increase survival for many patients for whom current treatments are not effective. However, response rates to current immune checkpoint inhibitors vary widely between patients and different types of cancer, and the mechanisms underlying these varied responses are poorly understood. Insights into the antitumor activities of checkpoint inhibitors are often obtained using syngeneic mouse models, which provide an in vivo preclinical basis for predicting efficacy in human clinical trials. Efforts to establish in vitro syngeneic mouse equivalents, which could increase throughput and permit real-time evaluation of lymphocyte infiltration and tumor killing, have been hampered by difficulties in recapitulating the tumor microenvironment in laboratory systems. Here, we describe a multiplex in vitro system that overcomes many of the deficiencies seen in current static histocultures, which we applied to the evaluation of checkpoint blockade in tumors derived from syngeneic mouse models. Our system enables both precision-controlled perfusion across biopsied tumor fragments and the introduction of checkpoint-inhibited tumor-infiltrating lymphocytes in a single experiment. Through real-time high-resolution confocal imaging and analytics, we demonstrated excellent correlations between in vivo syngeneic mouse and in vitro tumor biopsy responses to checkpoint inhibitors, suggesting the use of this platform for higher throughput evaluation of checkpoint efficacy as a tool for drug development.

scholarly journals Targeting the COX2/MET/TOPK signaling axis induces apoptosis in gefitinib-resistant NSCLC cells

2019 ◽  
Vol 10 (10) ◽  
Author(s):  
Juanjuan Xiao ◽  
Fei Wang ◽  
Hui Lu ◽  
Sanpeng Xu ◽  
Ling Zou ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Ex Vivo ◽  
Low Cost ◽  
Progression Free Survival ◽  
Signaling Axis ◽  
Approved Drugs ◽  
Gefitinib Resistance ◽  
Resistant Cells

Abstract MET overactivation is one of the crucial reasons for tyrosine kinase inhibitor (TKI) resistance, but the mechanisms are not wholly clear. Here, COX2, TOPK, and MET expression were examined in EGFR-activating mutated NSCLC by immunohistochemical (IHC) analysis. The relationship between COX2, TOPK, and MET was explored in vitro and ex vivo. In addition, the inhibition of HCC827GR cell growth by combining COX2 inhibitor (celecoxib), TOPK inhibitor (pantoprazole), and gefitinib was verified ex vivo and in vivo. We found that COX2 and TOPK were highly expressed in EGFR-activating mutated NSCLC and the progression-free survival (PFS) of triple-positive (COX2, MET, and TOPK) patients was shorter than that of triple-negative patients. Then, we observed that the COX2-TXA2 signaling pathway modulated MET through AP-1, resulting in an inhibition of apoptosis in gefitinib-resistant cells. Moreover, we demonstrated that MET could phosphorylate TOPK at Tyr74 and then prevent apoptosis in gefitinib-resistant cells. In line with these findings, the combination of celecoxib, pantoprazole, and gefitinib could induce apoptosis in gefitinib-resistant cells and inhibit tumor growth ex vivo and in vivo. Our work reveals a novel COX2/MET/TOPK signaling axis that can prevent apoptosis in gefitinib-resistant cells and suggests that a triple combination of FDA-approved drugs would provide a low-cost and practical strategy to overcome gefitinib resistance.

scholarly journals The RNA Methyltransferase METTL3 Promotes Endothelial Progenitor Cell Angiogenesis in Mandibular Distraction Osteogenesis via the PI3K/AKT Pathway

2021 ◽  
Vol 9 ◽  
Author(s):  
Weidong Jiang ◽  
Peiqi Zhu ◽  
Fangfang Huang ◽  
Zhenchen Zhao ◽  
Tao Zhang ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Distraction Osteogenesis ◽  
Ex Vivo ◽  
Maxillofacial Surgery ◽  
Akt Pathway ◽  
Oral And Maxillofacial Surgery ◽  
Endothelial Progenitor ◽  
Mandibular Distraction Osteogenesis

Distraction osteogenesis (DO) is used to treat large bone defects in the field of oral and maxillofacial surgery. Successful DO-mediated bone regeneration is dependent upon angiogenesis, and endothelial progenitor cells (EPCs) are key mediators of angiogenic processes. The N6-methyladenosine (m6A) methyltransferase has been identified as an important regulator of diverse biological processes, but its role in EPC-mediated angiogenesis during DO remains to be clarified. In the present study, we found that the level of m6A modification was significantly elevated during the process of DO and that it was also increased in the context of EPC angiogenesis under hypoxic conditions, which was characterized by increased METTL3 levels. After knocking down METTL3 in EPCs, m6A RNA methylation, proliferation, tube formation, migration, and chicken embryo chorioallantoic membrane (CAM) angiogenic activity were inhibited, whereas the opposite was observed upon the overexpression of METTL3. Mechanistically, METTL3 silencing reduced the levels of VEGF and PI3Kp110 as well as the phosphorylation of AKT, whereas METTL3 overexpression reduced these levels. SC79-mediated AKT phosphorylation was also able to restore the angiogenic capabilities of METTL3-deficient EPCs in vitro and ex vivo. In vivo, METTL3-overexpressing EPCs were additionally transplanted into the DO callus, significantly enhancing bone regeneration as evidenced by improved radiological and histological manifestations in a canine mandibular DO model after consolidation over a 4-week period. Overall, these results indicate that METTL3 accelerates bone regeneration during DO by enhancing EPC angiogenesis via the PI3K/AKT pathway.

scholarly journals Allogeneic Bone-Marrow Mesenchymal Stem Cell with Moldable Cryogel for Craniofacial Bone Regeneration

2021 ◽  
Vol 11 (12) ◽  
pp. 1326
Author(s):  
Cheng-Feng Chu ◽  
Shih-Hsuan Mao ◽  
Victor Bong-Hang Shyu ◽  
Chih-Hao Chen ◽  
Chien-Tzung Chen
Keyword(s):  
Bone Marrow ◽  
Bone Regeneration ◽  
Ex Vivo ◽  
Cell Lineage ◽  
Allogeneic Bone Marrow ◽  
Allogeneic Bone ◽  
Micro Computed Tomography ◽  
Alternative Source

Allogeneic bone-marrow mesenchymal stem cells (BMSCs) can promote bone regeneration and substitute for autologous BMSCs if autologous sources are unavailable, but the efficacy of bone regeneration by allogeneic BMSCs is still inconsistent. A Lewis rat cranium defect model was used to investigate the efficacy of bone regeneration between autologous and allogeneic BMSCs in gelatin-nanohydroxyapatite cryogel scaffolds. BMSCs from Wistar rats served as the allogeneic cell lineage. The full-thickness cranium defects were treated by either blank control, cryogel only, allogeneic BMSC-seeded cryogel, or autologous BMSC-seeded cryogel (n = 5). Bone regeneration was monitored by micro-computed tomography and examined histologically at week 12. In addition, we assessed the immune responses in vitro by mixed lymphocyte reaction (MLR) assay and CD4+ immunochemistry staining ex vivo. The MLR showed that allogeneic BSMCs elicited a weak immune response on day 14 that progressively attenuated by day 28. In vivo, the bone regeneration in allogeneic BMSCs was inferior at week 4, but progressively matched the autologous BMSCs by week 12. Our results suggest that allogeneic BMSCs can serve as an alternative source for bone regeneration.

scholarly journals Cochlear Dummy Electrodes for Insertion Training and Research Purposes: Fabrication, Mechanical Characterization, and Experimental Validation

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Jan-Philipp Kobler ◽  
Anandhan Dhanasingh ◽  
Raphael Kiran ◽  
Claude Jolly ◽  
Tobias Ortmaier
Keyword(s):  
Mechanical Properties ◽  
Force Measurement ◽  
Ex Vivo ◽  
Low Cost ◽  
Hearing Preservation ◽  
Insertion Force ◽  
A New Technique ◽  
Temporal Bones

To develop skills sufficient for hearing preservation cochlear implant surgery, surgeons need to perform several electrode insertion trials inex vivotemporal bones, thereby consuming relatively expensive electrode carriers. The objectives of this study were to evaluate the insertion characteristics of cochlear electrodes in a plastic scala tympani model and to fabricate radio opaque polymer filament dummy electrodes of equivalent mechanical properties. In addition, this study should aid the design and development of new cochlear electrodes. Automated insertion force measurement is a new technique to reproducibly analyze and evaluate the insertion dynamics and mechanical characteristics of an electrode. Mechanical properties of MED-EL’s FLEX28, FLEX24, and FLEX20electrodes were assessed with the help of an automated insertion tool. Statistical analysis of the overall mechanical behavior of the electrodes and factors influencing the insertion force are discussed. Radio opaque dummy electrodes of comparable characteristics were fabricated based on insertion force measurements. The platinum-iridium wires were replaced by polymer filament to provide sufficient stiffness to the electrodes and to eradicate the metallic artifacts in X-ray and computed tomography (CT) images. These low-cost dummy electrodes are cheap alternatives for surgical training and forin vitro, ex vivo, andin vivoresearch purposes.

scholarly journals Activated neutrophil fluorescent imaging technique for human lungs

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Thomas H. Craven ◽  
Tashfeen Walton ◽  
Ahsan R. Akram ◽  
Emma Scholefield ◽  
Neil McDonald ◽  
...  
Keyword(s):  
Real Time ◽  
Ex Vivo ◽  
Optical Signal ◽  
Neutrophil Activation ◽  
Fluorescent Imaging ◽  
Human Neutrophil Elastase ◽  
Clinical Criteria ◽  
Integral Process

AbstractNeutrophil activation is an integral process to acute inflammation and is associated with adverse clinical sequelae. Identification of neutrophil activation in real time in the lungs of patients may permit biological stratification of patients in otherwise heterogenous cohorts typically defined by clinical criteria. No methods for identifying neutrophil activation in real time in the lungs of patients currently exist. We developed a bespoke molecular imaging probe targeting three characteristic signatures of neutrophil activation: pinocytosis, phagosomal alkalinisation, and human neutrophil elastase (HNE) activity. The probe functioned as designed in vitro and ex vivo. We evaluated optical endomicroscopy imaging of neutrophil activity using the probe in real-time at the bedside of healthy volunteers, patients with bronchiectasis, and critically unwell mechanically ventilated patients. We detected a range of imaging responses in vivo reflecting heterogeneity of condition and severity. We corroborated optical signal was due to probe function and neutrophil activation.

scholarly journals Refinement of a mouse cardiovascular model: Development, application and dissemination

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 593
Author(s):  
Kirk A. Taylor ◽  
Michael Emerson
Keyword(s):  
Real Time ◽  
Light Transmission ◽  
Ex Vivo ◽  
Model Development ◽  
Cell Counting ◽  
Real Time Monitoring ◽  
Mortality Models ◽  
Typical Experiment

European and UK legislation requires all animal procedures to be conducted with consideration to reduction, refinement and replacement. In this review, 3Rs developments are discussed in the field of platelet biology and thromboembolism. Platelet research requires the use of animal models, and mice are widely used in the field. When working in vitro, conventional light transmission techniques have been scaled down allowing reduction in animal numbers. In vivo, vascular injury models are widely used and work is ongoing to develop ex vivo approaches that use fewer animals. Thromboembolic mortality models, which inflict considerable pain and suffering, have also been used widely. A published and characterised refinement of this mortality model allows real-time monitoring of radiolabelled platelets under general anaesthesia and reduces both the severity level and the numbers of mice used in a typical experiment. This technique is more sensitive than the mortality approach and has opened up new avenues of research, which would not have been feasible by using death as an end-point. To drive uptake of real-time monitoring, a more simplistic approach has been developed involving micro-sampling and cell counting. Thromboembolic mortality models should therefore be considered obsolete due to the emergence of 3Rs models with improved scientific outcomes and that can be implemented relatively easily.

scholarly journals Applications of novel bioreactor technology to enhance the viability and function of cultured cells and tissues

2020 ◽  
Vol 10 (2) ◽  
pp. 20190090 ◽  
Author(s):  
H. W. Hoyle ◽  
L. A. Smith ◽  
R. J. Williams ◽  
S. A. Przyborski
Keyword(s):  
Cultured Cells ◽  
Ex Vivo ◽  
Low Cost ◽  
Tissue Slices ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Tissue Constructs ◽  
And Function

As the field of tissue engineering continues to advance rapidly, so too does the complexity of cell culture techniques used to generate in vitro tissue constructs, with the overall aim of mimicking the in vivo microenvironment. This complexity typically comes at a cost with regards to the size of the equipment required and associated expenses. We have developed a small, low-cost bioreactor system which overcomes some of the issues of typical bioreactor systems while retaining a suitable scale for the formation of complex tissues. Herein, we have tested this system with three cell populations/tissues: the culture of hepatocellular carcinoma cells, where an improved structure and basic metabolic function is seen; the culture of human pluripotent stem cells, in which the cultures can form more heterogeneous tissues resembling the in vivo teratoma and ex vivo liver tissue slices, in which improved maintenance of cellular viability is seen over the 3 days tested. This system has the flexibility to be used for a variety of further uses and has the potential to provide a more accessible alternative to current bioreactor technologies.

In-Vivo Real-Time Contrast-Free Ultrasonic Blood Flow Velocity Profile Measurement

2012 ◽  
Author(s):  
G. G. Koutsouridis ◽  
N. Bijnens ◽  
P. J. Brands ◽  
F. N. van de Vosse ◽  
M. C. M. Rutten
Keyword(s):  
Velocity Profile ◽  
Real Time ◽  
Ex Vivo ◽  
Accurate Estimation ◽  
Profile Measurement ◽  
Processing Unit ◽  
Local Pressure ◽  
Measured Velocity

Recently, Ultrasonic Perpendicular Velocimetry (UPV) based algorithms, as opposed to commonly used Doppler technique (Figure 1), were applied to Radio Frequency (RF)-data acquired in an in-vitro setup [1,3]. Thus, the estimation of velocity components perpendicularly to the ultrasound beam and the simultaneous and accurate assessment of wall position and axial velocity profiles were made feasible. By integrating the measured velocity profile an accurate flow estimation was made possible. Furthermore, the ratio between the changes in flow Q(t) and the changes in cross-sectional area of the vessel A(t) was found to offer an accurate estimation of the local Pulse Wave Velocity (PWV). By combining the PWV with the diameter waveform, accurate local pressure estimation was obtained indicating that a non-invasive pressure assessment by means of ultrasound is feasible [3]. However, the abovementioned method is time consuming due to the data size and the post-processing procedure required. Additionally, the Fast Fourier Transform (FFT) on Butterworth Band Pass Filters (BPF) for vessel’s wall removal requires contrast agents dispersion in the fluid for the application of UPV. A real-time approach, of the previously described techniques, was applied [2] in-vitro using a Blood Mimicking Fluid (BMF), as contrast agent, resembling the rheological (shear thinning) and acoustical (backscattering) properties of blood and ex-vivo using BMF or contrast-free real blood implementing Wavelet Transform (WT) filtering. The use of a Graphics Processing Unit (GPU) [4], succeeded in considerable acceleration and WT [5] filtering on the rough RF-data, in improvement of the discrimination between reflections from the vessel wall and scattering from small particles. In this research the method is extended to include in-vivo measurements.

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