IMPROVING THE CLINICAL UTILITY OF MESENCHYMAL STEM CELLS THROUGH ENHANCING CELL SURVIVAL: ENGINEERED AND CELLULAR APPROACHES

For decades, mesenchymal stem cells have been touted as highly promising cells for use in regenerative medicine. Their flexibility in differentiation to other cell types, paracrine abilities in supporting angiogenesis, and modulatory behavior in inflammation have led to researchers applying the cells across a wide variety of treatments, including skin healing, bone regeneration, myocardial infarction repair, and others. However, a fundamental limitation in all MSC therapies has been a lack of survival for implanted cells, preventing them from carrying out their many beneficial functions. We have examined strategies to improve this MSC survival dilemma, hypothesizing that a combination of engineered approaches to activate survival signaling and modulation of autophagy to generate energy for the cell would act to improve MSC survival and function in the face of stressors. Our results show that cell surface restriction of EGFR to constitutively activate downstream survival signaling improves MSC longevity in two systems: physical tethering of soluble EGF on bone regeneration scaffolds, and EGFR tethering through EGF-like repeats of the ECM protein Tenascin C in a polymer for skin wound healing. Additionally, we found that MSCs have a high accumulation of autophagosomes, which are mobilized for degradation during the stress of differentiation and can be tuned to affect MSC function in relation to differentiation. Ultimately, these strategies to alter MSC signaling and function will be of considerable use for attempts to apply the great potential utility of MSCs to wound healing and other contexts in biomedical science

Advanced Testing Chain Supporting the Validation of Smart Grid Systems and Technologies

New testing and development procedures and methods are needed to address topics like power system stability, operation and control in the context of grid integration of rapidly developing smart grid technologies. In this context, individual testing of units and components has to be reconsidered and appropriate testing procedures and methods need to be described and implemented. This paper addresses these needs by proposing a holistic and enhanced testing methodology that integrates simulation/software- and hardware-based testing infrastructure. This approach presents the advantage of a testing environment, which is very close to f i eld testing, includes the grid dynamic behavior feedback and is risks-free for the power system, for the equipment under test and for the personnel executing the tests. Furthermore, this paper gives an overview of successful implementation of the proposed testing approach within different testing infrastructure available at the premises of different research institutes in Europe.Comment: 2018 IEEE Workshop on Complexity in Engineering (COMPENG

Technology in Practice: Promoting Participation in Patients with High Level of Spinal Cord Injury

Motor impairment often occurs following a spinal cord injury, limiting participation in functional tasks. Although the primary rehabilitative focus is to cure paralysis, not all patients experience full recovery. For those who may not regain complete motor function, rehabilitative interventions are needed to bridge the gap between disability and optimal participation. During rehabilitation, patients prioritize increasing independence and participation in meaningful activities (Rigby, Ryan, & Campbell, 2010). More specifically, regaining arm and hand function was reported as a top priority during treatment for individuals with complete spinal cord injury (Peckham, et al., 2001). Technology has been utilized to improve participation in individuals with tetraplegia, who may only regain partial function (Ripat & Woodgate, 2012). A review of current evidence was conducted to determine technological devices that are used to increase autonomy in individuals with high level spinal cord injury. The purpose of this session is to present an evidence based review focused on various modes of technologies that can be used both during the rehabilitation process and in the home. A search of PubMed and CINAHL databases yielded approximately 1,200 resources, which were refined to sixteen journal articles that included evaluations of upper extremity interventions for adults and adolescents with cervical and thoracic level spinal cord. The selected articles for this review primarily focused on functional electrical stimulation, such as brain-computer-interface and neuroprostheses, as well as electronic aids of daily living. Articles were critically reviewed by two raters. Overall, individuals experienced increased participation when utilizing technology to perform functional tasks. Occupational therapists play an integral role in the implementation and patient-training of these devices to increase function and participation in daily activities. Examples of occupations in which technology has been integrated range from feeding and hygiene management to leisure pursuits. As technology continues to advance, therapists working with this population will need to increase their knowledge of these technologies and how to incorporate them into therapeutic interventions. Although existing evidence supports the use of these devices for facilitating function, there is a need for more high level evidence to further establish technology as an effective intervention with this population. References: Peckham, P. H., Keith, M. W., Kilgore, K. L., Grill, J. H., Wuolle, K. S., Thrope, G. B., ... & Wiegner, A. (2001). Efficacy of an implanted neuroprosthesis for restoring hand grasp in tetraplegia: a multicenter study. Archives of physical medicine and rehabilitation, 82(10), 1380-1388. Rigby, P., Ryan, S., & Campbell, K., A. (2011). Electronic aids to daily living and quality of life for persons with tetraplegia. Disability & Rehabilitation: Assistive Technology, 6(3), 260-267. doi:10.3109/17483107.2010.522678 Ripat, J. D., & Woodgate, R. L. (2012). Self-perceived participation among adults with spinal cord injury: A grounded theory study. Spinal cord, 50, 908-914

Human mesenchymal stem cells/multipotent stromal cells consume accumulated autophagosomes early in differentiation

Introduction: Bone marrow mesenchymal stem cells/multipotent stromal cells (MSCs) are recruited to sites of injury and subsequently support regeneration through differentiation or paracrine activity. During periods of stress such as wound site implant or differentiation, MSCs are subjected to a variety of stressors that might activate pathways to improve cell survival and generate energy. In this study, we monitored MSC autophagy in response to the process of differentiation. Methods: MSC autophagosome structures were observed by using transmission electron microscopy and a tandem green fluorescent protein-red fluorescent protein autophagic flux reporter to monitor the mammalian microtubule-associated protein-1 light chain 3 (LC3) turnover in real time. MSCs were differentiated by using standard osteogenic and adipogenic media, and autophagy was examined during short-term and long-term differentiation via immunoblots for LC3I and II. Autophagy was modulated during differentiation by using rapamycin and bafilomycin treatments to disrupt the autophagosome balance during the early stages of the differentiation process, and differentiation was monitored in the long term by using Von Kossa and Oil Red O staining as well as quantitative polymerase chain reaction analysis of typical differentiation markers. Results: We found that undifferentiated MSCs showed an accumulation of a large number of undegraded autophagic vacuoles, with little autophagic turnover. Stimulation of autophagy with rapamycin led to rapid degradation of these autophagosomes and greatly increased rough endoplasmic reticulum size. Upon induction of osteogenic differentiation, MSC expression of LC3II, a common autophagosome marker, was lost within 12 hours, consistent with increased turnover. However, during adipogenic differentiation, drug treatment to alter the autophagosome balance during early differentiation led to changes in differentiation efficiency, with inhibited adipocyte formation following rapamycin treatment and accelerated fat accumulation following autophagosome blockade by bafilomycin. Conclusions: Our findings suggest that MSCs exist in a state of arrested autophagy with high autophagosome accumulation and are poised to rapidly undergo autophagic degradation. This phenotype is highly sensitive, and a balance of autophagy appears to be key in efficient MSC differentiation and function, as evidenced by our results implicating autophagic flux in early osteogenesis and adipogenesis

Patient Frailty: Key Considerations, Definitions and Practical Implications

By 2020, the elderly (≥65-year-old) world population is projected to exceed one billion individuals. This demographic megatrend has brought topics such as physiological age and frailty to the forefront of medical research efforts around the globe. The concept of frailty has evolved significantly since the mid-twentieth century. The outdated stereotype of a “thin, stooped, slow octogenarian” has transitioned to a more scientific and objective understanding of the problem. Still, a comprehensive and concise definition of “frailty” remains elusive. Until such a definition is firmly established and universally agreed upon, clinicians continue to rely on the somewhat subjective conceptual framework of today. In this chapter, the authors review key issues pertaining to clinical management of frail patients, including diagnosis/identification, preventive strategies, therapeutic approaches, and common pitfalls. The relationship between frailty, various domains of life, and functional status is also discussed. Finally, we will touch upon the concepts of end-of-life and goals of care, focusing on their relationship to frailty

Longitudinal In Vivo Imaging of Retinal Ganglion Cells and Retinal Thickness Changes Following Optic Nerve Injury in Mice

Retinal ganglion cells (RGCs) die in sight-threatening eye diseases. Imaging RGCs in humans is not currently possible and proof of principle in experimental models is fundamental for future development. Our objective was to quantify RGC density and retinal thickness following optic nerve transection in transgenic mice expressing cyan fluorescent protein (CFP) under control of the Thy1 promoter, expressed by RGCs and other neurons.A modified confocal scanning laser ophthalmoscopy (CSLO)/spectral-domain optical coherence tomography (SD-OCT) camera was used to image and quantify CFP+ cells in mice from the B6.Cg-Tg(Thy1-CFP)23Jrs/J line. SD-OCT circle (1 B-scan), raster (37 B-scans) and radial (24 B-scans) scans of the retina were also obtained. CSLO was performed at baseline (n = 11) and 3 (n = 11), 5 (n = 4), 7 (n = 10), 10 (n = 6), 14 (n = 7) and 21 (n = 5) days post-transection, while SD-OCT was performed at baseline and 7, 14 and 35 days (n = 9) post-transection. Longitudinal change in CFP+ cell density and retinal thickness were computed. Compared to baseline, the mean (SD) percentage CFP+ cells remaining at 3, 5, 7, 10, 14 and 21 days post-transection was 86 (9)%, 63 (11)%, 45 (11)%, 31 (9)%, 20 (9)% and 8 (4)%, respectively. Compared to baseline, the mean (SD) retinal thickness at 7 days post-transection was 97 (3)%, 98 (2)% and 97 (4)% for the circle, raster and radial scans, respectively. The corresponding figures at 14 and 35 days post-transection were 96 (3)%, 97 (2)% and 95 (3)%; and 93 (3)%, 94 (3)% and 92 (3)%.Longitudinal imaging showed an exponential decline in CFP+ cell density and a small (≤8%) reduction in SD-OCT measured retinal thickness post-transection. SD-OCT is a promising tool for detecting structural changes in experimental optic neuropathy. These results represent an important step towards translation for clinical use