A High Intensity Focused Ultrasound System for Veterinary Oncology Applications.

BACKGROUND: Magnetic resonance-guided focused ultrasound surgery is an incisionless energy-based thermal method that is used for ablating tumors in the veterinary clinic. AIMS AND OBJECTIVES: In this article we describe a prototype of a veterinary system compatible with magnetic resonance imaging intended for small-to-medium-sized companion animals that was developed and tested in vivo in adult rabbits. METHODS: Real-time monitoring of the ablation during the experiment was possible with MR thermometry. Experiments involved thermal monitoring of sonications applied in the thigh of the rabbits. A 38-mm diameter transducer operating at 2.6 MHz was used with a 60-mm-focal length. The robotic system employed 3 linear axes and one angular axis. For this study, only X and Y axis were enabled. Due to the target size limitations, motion in Z and Θ was not needed. The functionality of the positioning device was evaluated by means of MR thermometry, demonstrating sufficient heating and accurate motion in both axes of operation. RESULTS: The postmortem findings confirm the ability of the system to induce thermal ablations in vivo in the absence of adverse effects. CONCLUSIONS: The device is a reliable and affordable solution for companion animal hospitals, offering and additional tool for the veterinary oncology society.

Scaffold and scaffold-free self-assembled systems in regenerative medicine.

Self-assembly in tissue engineering refers to the spontaneous chemical or biological association of components to form a distinct functional construct, reminiscent of native tissue. Such self-assembled systems have been widely used to develop platforms for the delivery of therapeutic and/or bioactive molecules and various cell populations. Tissue morphology and functional characteristics have been recapitulated in several self-assembled constructs, designed to incorporate stimuli responsiveness and controlled architecture through spatial confinement or field manipulation. In parallel, owing to substantial functional properties, scaffold-free cell-assembled devices have aided in the development of functional neotissues for various clinical targets. Herein, we discuss recent advancements and future aspirations in scaffold and scaffold-free self-assembled devices for regenerative medicine purposes. Biotechnol. Bioeng. 2016;113: 1155-1163. © 2015 Wiley Periodicals, Inc.

MRI compatibility testing of commercial high intensity focused ultrasound transducers.

PURPOSE: The study aimed to compare the performance of eight commercially available single-element High Intensity Focused Ultrasound (HIFU) transducers in terms of Magnetic Resonance Imaging (MRI) compatibility. METHODS: Imaging of an agar-based MRI phantom was performed in a 3 T MRI scanner utilizing T2-Weighted Fast Spin Echo (FSE) and Fast low angle shot (FLASH) sequences, which are typically employed for high resolution anatomical imaging and thermometry, respectively. Reference magnitude and phase images of the phantom were compared with images acquired in the presence of each transducer in terms of the signal to noise ratio (SNR), introduced artifacts, and overall image quality. RESULTS: The degree of observed artifacts highly differed among the various transducers. The transducer whose backing material included magnetic impurities showed poor performance in the MRI, introducing significant susceptibility artifacts such as geometric distortions and signal void bands. Additionally, it caused the most significant SNR drop. Other transducers were shown to exhibit high level of MRI compatibility as the resulting images closely resembled the reference images with minimal to no apparent artifacts and comparable SNR values. CONCLUSIONS: The study findings may facilitate researchers to select the most suitable transducer for their research, simultaneously avoiding unnecessary testing. The study further provides useful design considerations for MRI compatible transducers.

ß-Tocotrienol and δ-Tocotrienol as Additional Inhibitors of the Main Protease of Feline Infectious Peritonitis Virus: An In Silico Analysis.

Feline infectious peritonitis (FIP) is a severe and invariably fatal disease affecting both domestic and wild felines with limited effective therapeutic options available. By considering the significant immunomodulatory effects of vitamin E observed in both animal and human models under physiological and pathological conditions, we have provided a full in silico investigation of vitamin E and related compounds and their effect on the crystal structure of feline infectious peritonitis virus 3C-like protease (FIPV-3CLpro). This work revealed the ß-tocotrienol and δ-tocotrienol analogs as inhibitor candidates for this protein, suggesting their potential as possible drug compounds against FIP or their supplementary use with current medicines against this disease.

Effect of Magnetic Resonance Imaging on the Motion Accuracy of Magnetic Resonance Imaging-compatible Focused Ultrasound Robotic System.

Purpose: The current study provides insights into the challenges of safely operating a magnetic resonance imaging (MRI)-guided focused ultrasound (MRgFUS) robotic system in a high-field MRI scanner in terms of robotic motion accuracy. Materials and Methods: Grid sonications were carried out in phantoms and excised porcine tissue in a 3T MRI scanner using an existing MRgFUS robotic system. Fast low-angle shot-based magnetic resonance thermometry was employed for the intraprocedural monitoring of thermal distribution. Results: Strong shifting of the heated spots from the intended points was observed owing to electromagnetic interference (EMI)-induced malfunctions in system's operation. Increasing the slice thickness of the thermometry sequence to at least 8 mm was proven an efficient method for preserving the robotic motion accuracy. Conclusions: These findings raise awareness about EMI effects on the motion accuracy of MRgFUS robotic devices and how they can be mitigated by employing suitable thermometry parameters.

Treatment of mammary cancer with focused ultrasound: A pilot study in canine and feline patients.

In recent years, veterinary medicine has expanded its practices beyond conventional methods, gradually integrating the Focused Ultrasound (FUS) technology in the care of companion animals like dogs and cats. The current study aimed to examine the feasibility and provide insights into the application of thermal FUS in canine and feline mammary cancer therapy. FUS was delivered by a 2-MHz single-element spherically focused ultrasonic transducer as integrated with an existing robotic positioning device. The functionality of the FUS system and sonication protocol in efficiently and safely ablating live tissue was initially validated in a rabbit thigh model in a laboratory environment. Nine (9) dogs and cats with superficial mammary cancer were recruited through a dedicated campaign according to specific safety criteria. The veterinary patients underwent FUS ablation followed by immediate surgical resection of the entire malignancy. Histopathology examination demonstrated well-defined regions of coagulative necrosis in all treated tumors with no off-target damage. Further study with a larger patient population is needed to confirm the current findings and demonstrate the safety and feasibility of complete FUS ablation of deep-seated tumors.

Treatment of canine and feline sarcoma using MR-guided focused ultrasound system.

PURPOSE: In recent years, veterinary medicine has enhanced its applications beyond traditional approaches, progressively incorporating the Focused Ultrasound (FUS) technology. This study investigated the ability of FUS to precisely ablate naturally occurring canine and feline soft tissue sarcomas (STS). METHODS: Six dogs and four cats with superficial tumours were enrolled in the study. The tumours were treated with a Magnetic Resonance guided FUS (MRgFUS) robotic system featuring a single element spherically focused transducer of 2.6 MHz. The tumours were then removed by surgery and sent for hematoxylin and eosin (H&E) staining. RESULTS: The MRgFUS system was capable of inflicting well-defined overlapping lesions in the tumours. The anatomical sites of the treated tumours were the neck, leg, face, back and belly. Coagulative necrosis was evidenced by histopathology assessment in 80% of cases. CONCLUSION: Therefore, this technology can be a therapeutic solution for veterinary cancer and a model for advancing the knowledge on human STS.

Robotic system for magnetic resonance guided focused ultrasound ablation of abdominal cancer.

BACKGROUND: A prototype robotic system that uses magnetic resonance guided focused ultrasound (MRgFUS) technology is presented. It features three degrees of freedom (DOF) and is intended for thermal ablation of abdominal cancer. METHODS: The device is equipped with three identical transducers being offset between them, thus focussing at different depths in tissue. The efficacy and safety of the system in ablating rabbit liver and kidney was assessed, both in laboratory and magnetic resonance imaging (MRI) conditions. RESULTS: Despite these organs' challenging location, in situ coagulative necrosis of a tissue area was achieved. Heating of abdominal organs in rabbit was successfully monitored with MR thermometry. CONCLUSIONS: The MRgFUS system was proven successful in creating lesions in the abdominal area of rabbits. The outcomes of the study are promising for future translation of the technology to the clinic.

Magnetic resonance image-guided focused ultrasound robotic system for transrectal prostate cancer therapy.

BACKGROUND: A magnetic resonance image (MRI) guided robotic device for focussed ultrasound therapy of prostate cancer (PC) was developed. The device offers movement in 5 degrees of freedom (DOF) and uses a single-element transducer that operates at 3.2 MHz, has a diameter of 25 mm and focuses at 45 mm. METHODS: The MRI compatibility of the system was evaluated in a 1.5 T scanner. The ability of the transducer to create lesions was evaluated in laboratory and MRI settings, on ex vivo pork tissue and in vivo rabbit thigh tissue. RESULTS: Cavitational and thermal lesions were created on the excised pork tissue. In vivo experiments proved the efficacy of the system in ablating muscle tissue without damaging intervening areas. CONCLUSIONS: The MRI compatible robotic system can be placed on the table of any commercial MRI scanner up to 7 T. The device has the ability of future use for transrectal focal therapy of PC with the patient in supine position.

The synergistic effect of low oxygen tension and macromolecular crowding in the development of extracellular matrix-rich tendon equivalents.

Cellular therapies play an important role in tendon tissue engineering, with tenocytes being the most prominent and potent cell population available. However, for the development of a rich extracellular matrix tenocyte-assembled tendon equivalent, prolonged in vitro culture is required, which is associated with phenotypic drift. Recapitulation of tendon tissue microenvironment in vitro with cues that enhance and accelerate extracellular matrix synthesis and deposition, whilst maintaining tenocyte phenotype, may lead to functional cell therapies. Herein, we assessed the synergistic effect of low oxygen tension (enhances extracellular matrix synthesis) and macromolecular crowding (enhances extracellular matrix deposition) in human tenocyte culture. Protein analysis demonstrated that human tenocytes at 2% oxygen tension and with 50 µg ml-1 carrageenan (macromolecular crowder used) significantly increased synthesis and deposition of collagen types I, III, V and VI. Gene analysis at day 7 illustrated that human tenocytes at 2% oxygen tension and with 50 µg ml-1 carrageenan significantly increased the expression of prolyl 4-hydroxylase subunit alpha 1, procollagen-lysine 2- oxoglutarate 5-dioxygenase 2, scleraxis, tenomodulin and elastin, whilst chondrogenic (e.g. runt-related transcription factor 2, cartilage oligomeric matrix protein, aggrecan) and osteogenic (e.g. secreted phosphoprotein 1, bone gamma-carboxyglutamate protein) trans-differentiation markers were significantly down-regulated or remained unchanged. Collectively, our data clearly illustrates the beneficial synergistic effect of low oxygen tension and macromolecular crowding in the accelerated development of tissue equivalents.

Progress in cell-based therapies for tendon repair.

The last decade has seen significant developments in cell therapies, based on permanently differentiated, reprogrammed or engineered stem cells, for tendon injuries and degenerative conditions. In vitro studies assess the influence of biophysical, biochemical and biological signals on tenogenic phenotype maintenance and/or differentiation towards tenogenic lineage. However, the ideal culture environment has yet to be identified due to the lack of standardised experimental setup and readout system. Bone marrow mesenchymal stem cells and tenocytes/dermal fibroblasts appear to be the cell populations of choice for clinical translation in equine and human patients respectively based on circumstantial, rather than on hard evidence. Collaborative, inter- and multi-disciplinary efforts are expected to provide clinically relevant and commercially viable cell-based therapies for tendon repair and regeneration in the years to come.

Data on in vitro and in vivo cell orientation on substrates with different topographies.

This data article contains data related to the research article entitled "Substrate topography: A valuable in vitro tool, but a clinical red herring for in vivo tenogenesis" [1]. We report measurements on tenocyte viability, metabolic activity and proliferation on substrates with different topographies. We also report the effect of substrates with different topographies on host cells in a subcutaneous model.

Substrate topography: A valuable in vitro tool, but a clinical red herring for in vivo tenogenesis.

Controlling the cell-substrate interactions at the bio-interface is becoming an inherent element in the design of implantable devices. Modulation of cellular adhesion in vitro, through topographical cues, is a well-documented process that offers control over subsequent cellular functions. However, it is still unclear whether surface topography can be translated into a clinically functional response in vivo at the tissue/device interface. Herein, we demonstrated that anisotropic substrates with a groove depth of ∼317nm and ∼1988nm promoted human tenocyte alignment parallel to the underlying topography in vitro. However, the rigid poly(lactic-co-glycolic acid) substrates used in this study upregulated the expression of chondrogenic and osteogenic genes, indicating possible tenocyte trans-differentiation. Of significant importance is that none of the topographies assessed (∼37nm, ∼317nm and ∼1988nm groove depth) induced extracellular matrix orientation parallel to the substrate orientation in a rat patellar tendon model. These data indicate that two-dimensional imprinting technologies are useful tools for in vitro cell phenotype maintenance, rather than for organised neotissue formation in vivo, should multifactorial approaches that consider both surface topography and substrate rigidity be established. STATEMENT OF SIGNIFICANCE: Herein, we ventured to assess the influence of parallel groves, ranging from nano- to micro-level, on tenocytes response in vitro and on host response using a tendon and a subcutaneous model. In vitro analysis indicates that anisotropically ordered micro-scale grooves, as opposed to nano-scale grooves, maintain physiological cell morphology. The rather rigid PLGA substrates appeared to induce trans-differentiation towards chondrogenic and/or steogenic lineage, as evidence by TILDA gene analysis. In vivo data in both tendon and subcutaneous models indicate that none of the substrates induced bidirectional host cell and tissue growth. Collective, these observations indicate that two-dimensional imprinting technologies are useful tools for in vitro cell phenotype maintenance, rather than for directional neotissue formation, should multifactorial approaches that consider both surface topography and substrate rigidity be established.

The biophysical, biochemical, and biological toolbox for tenogenic phenotype maintenance in vitro.

Tendon injuries constitute an unmet clinical need, with 3 to 5 million new incidents occurring annually worldwide. Tissue grafting and biomaterial-based approaches fail to provide environments that are conducive to regeneration; instead they lead to nonspecific cell adhesion and scar tissue formation, which collectively impair functionality. Cell based therapies may potentially recover native tendon function, if tenocyte trans-differentiation can be evaded and stem cell differentiation towards tenogenic lineage is attained. To this end, recreating an artificial in vivo tendon niche by engineering functional in vitro microenvironments is a research priority. Clinically relevant cell based therapies for tendon repair and regeneration could be created using tools that harness biophysical beacons (surface topography, mechanical loading), biochemical cues (oxygen tension), and biological signals (growth factors).

Assessment of stem cell carriers for tendon tissue engineering in pre-clinical models.

Tendon injuries are prevalent and problematic, especially among young and otherwise healthy individuals. The inherently slow innate healing process combined with the inevitable scar tissue formation compromise functional recovery, imposing the need for the development of therapeutic strategies. The limited number of low activity/reparative capacity tendon-resident cells has directed substantial research efforts towards the exploration of the therapeutic potential of various stem cells in tendon injuries and pathophysiologies. Severe injuries require the use of a stem cell carrier to enable cell localisation at the defect site. The present study describes advancements that injectable carriers, tissue grafts, anisotropically orientated biomaterials, and cell-sheets have achieved in preclinical models as stem cell carriers for tendon repair.