Developing A New Strategy for Delivery of Neural Transplant Populations using Precursor Cell Sprays and Specialised Cell Media

Neural precursor/stem cell transplantation therapies promote regeneration in neurological injuries, but current cell delivery methods have drawbacks. These include risks with surgical microinjection (e.g., hemorrhage, embolism) and high cell loss with systemic delivery/passage through fine gauge needles. Aerosolized cell delivery offers significant benefits including rapid and minimally invasive cell delivery, and ease of delivery to end users. To develop this approach, it is necessary to prove that 1) aerosolization does not have detrimental effects on transplant cells and 2) suitable media can be identified to support cell delivery. To achieve these aims, cells are sprayed using a commercial spray device or stored in Hibernate-A, a CO2-independent nutrient solution. Histological assessments consist of cell viability analysis, immunocytochemistry, and EdU labeling. It is shown that a major neural precursor transplant population-oligodendrocyte precursor cells (OPCs)-survive following aerosolized delivery and retain their capacity for proliferation and differentiation (key to their repair function). Hibernate-A can support OPCs' survival without specialized maintenance conditions, with no detrimental impact on cell fate. It is considered that this data supports the concept of a novel class of advanced medical spray devices to facilitate transport and delivery of transplant populations in neural cell therapy

Developing human dish models of neurological pathology

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Safe nanoengineering and incorporation of transplant populations in a neurosurgical grade biomaterial, DuraGen PlusTM, for protected cell therapy applications

High transplant cell loss is a major barrier to translation of stem cell therapy for pathologies of the brain and spinal cord. Encapsulated delivery of stem cells in biomaterials for cell therapy is gaining popularity but experimental research has overwhelmingly used laboratory grade materials unsuitable for human clinical use representing a further barrier to clinical translation. A potential solution is to use neurosurgical grade materials routinely used in clinical protocols which have an established human safety profile. Here, we tested the ability of Duragen Plus (TM)- a clinical biomaterial used widely in neurosurgical duraplasty procedures, to support the growth and differentiation of neural stem cells- a major transplant population being tested in clinical trials for neurological pathology. Genetic engineering of stem cells yields augmented therapeutic cells, so we further tested the ability of the Duragen Plus (TM) matrix to support stem cells engineered using magnetofection technology and minicircle DNA vectors- a promising cell engineering approach we previously reported (Journal of Controlled Release, 2016 a &b). The safety of the nano-engineering approach was analysed for the first time using sophisticated data-independent analysis by mass spectrometry-based proteomics. We prove that the Duragen Plus (TM) matrix is a promising biomaterial for delivery of stem cell transplant populations, with no adverse effects on key regenerative parameters. This advanced cellular construct based on a combinatorial nano-engineering and biomaterial encapsulation approach, could therefore offer key advantages for clinical translation

Developing an Open-Source IoT Platform for Optimal Irrigation Scheduling and Decision-Making: Implementation at Olive Grove Parcels

Climate change has reduced the availability of good quality water for agriculture, while favoring the proliferation of harmful insects, especially in Mediterranean areas. Deploying IoT-based systems can help optimize water-use efficiency in agriculture and address problems caused by extreme weather events. This work presents an IoT-based monitoring system for obtaining soil moisture, soil electrical conductivity, soil temperature and meteorological data useful in irrigation management and pest control. The proposed system was implemented and evaluated for olive parcels located both at coastal and inland areas of the eastern part of Crete; these areas face severe issues with water availability and saltwater intrusion (coastal region). The system includes the monitoring of soil moisture and atmospheric sensors, with the aim of providing information to farmers for decision-making and at the future implementation of an automated irrigation system, optimizing the use of water resources. Data acquisition was performed through smart sensors connected to a microcontroller. Data were received at a portal and made available on the cloud, being monitored in real-time through an open-source IoT platform. An e-mail alert was sent to the farmers when soil moisture was lower than a threshold value specific to the soil type or when climatic conditions favored the development of the olive fruit fly. One of the main advantages of the proposed decision-making system is a low-cost IoT solution, as it is based on open-source software and the hardware on edge devices consists of widespread economic modules. The reliability of the IoT-based monitoring system has been tested and could be used as a support service tool offering an efficient irrigation and pest control service

In vitro model of traumatic brain injury to screen neuro-regenerative biomaterials

Penetrating traumatic brain injury (pTBI) causes serious neurological deficits with no clinical regenerative therapies currently available. Tissue engineering strategies using biomaterial-based ‘structural bridges’ offer high potential to promote neural regeneration post-injury. This includes surgical grade materials which can be repurposed as biological scaffolds to overcome challenges associated with long approval processes and scaleup for human application. However, high throughput, pathomimetic models of pTBI are lacking for the developmental testing of such neuro-materials, representing a bottleneck in this rapidly emergent field. We have established a high throughput and facile culture model containing the major neural cell types which govern biomaterial handling in the central nervous system. We show that induction of traumatic injuries was feasible in the model, with post-injury implantation of a surgical grade biomaterial. Cellular imaging in lesions was achievable using standard epifluorescence microscopy methods. Key pathological features of pTBI were evident in vitro namely immune cell infiltration of lesions/biomaterial, with responses characteristic of cell scarring, namely hypertrophic astrocytes with GFAP upregulation. Based on our observations, we consider the high-throughput, inexpensive and facile pTBI model can be used to study biomaterial ‘implantation’ and evaluate neural cell-biomaterial responses. The model is highly versatile to test a range of laboratory and clinical grade materials for neural regeneration

Subdural Effusions with Hydrocephalus after Severe Head Injury: Successful Treatment with Ventriculoperitoneal Shunt Placement: Report of 3 Adult Cases

Background. Subdural collections of cerebrospinal fluid (CSF) with associated hydrocephalus have been described by several different and sometimes inaccurate terms. It has been proposed that a subdural effusion with hydrocephalus (SDEH) can be treated effectively with a ventriculoperitoneal shunt (V-P shunt). In this study, we present our experience treating patients with SDEH without directly treating the subdural collection. Methods. We treated three patients with subdural effusions and hydrocephalus as a result of a head injury. All the patients were treated with a V-P shunt despite the fact that there was an extra-axial CSF collection with midline shift. Results. In all of the patients, the subdural effusions subsided and the ventricular dilatation improved in the postoperative period. The final clinical outcome remains difficult to predict and depends not only on the successful CSF diversion but also on the primary and secondary brain insult. Conclusion. Subdural effusions with hydrocephalus can be safely and effectively treated with V-P shunting, without directly treating the subdural effusion which subsides along with the treatment of hydrocephalus. However, it is extremely important to make an accurate diagnosis of an SDEH and differentiate this condition from other subdural collections which require different management

Stem cell sprays for neurological injuries: a perspective

Injuries to the brain and spinal cord have major clinical consequences with high costs for healthcare systems. Neural cell transplantation therapies have significant translational potential to promote regeneration post-injury with clinical trials commencing for various pathologies. However, there are challenges associated with current clinical approaches used for systemic or direct delivery of transplant cells to neural tissue in regenerative applications. These include risks associated with surgical microinjection into neural tissue (e.g. haemorrhage, cell clumping) and high cell loss due to systemic clearance or with cell passage through fine gauge needles into densely packed neural tissue. This article presents lines of evidence supporting the concept that cell spray delivery technology can offer significant translational benefits for neural transplantation therapy, versus current cell delivery methods. Potential benefits include rapid/homogenous cell delivery, release over large surface areas, minimal invasiveness, compatibility with neurosurgical procedures in acute injury, no predictable clinical complications and the capacity to combine cell therapies with drug/biomolecule delivery. Accordingly, we consider that the development of cell spray delivery technology represents a key goal to develop advanced cell therapies for regenerative neurology.</jats:p