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Dynamics of Zebrafish Heart Regeneration Using an HPLC-ESI-MS/MS Approach.
Ma D, Tu C, Sheng Q, Yang Y, Kan Z, Guo Y, Shyr Y, Scott IC, Lou X
(2018) J Proteome Res 17: 1300-1308
MeSH Terms: Animals, Chromatography, High Pressure Liquid, Fish Proteins, Gene Ontology, Heart Injuries, Heart Ventricles, Metabolic Networks and Pathways, Molecular Sequence Annotation, Myocardium, Proteomics, Real-Time Polymerase Chain Reaction, Regeneration, Spectrometry, Mass, Electrospray Ionization, Tumor Suppressor Protein p53, Zebrafish
Show Abstract · Added April 3, 2018
Failure to properly repair damaged due to myocardial infarction is a major cause of heart failure. In contrast with adult mammals, zebrafish hearts show remarkable regenerative capabilities after substantial damage. To characterize protein dynamics during heart regeneration, we employed an HPLC-ESI-MS/MS (mass spectrometry) approach. Myocardium tissues were taken from sham-operated fish and ventricle-resected sample at three different time points (2, 7, and 14 days); dynamics of protein expression were analyzed by an ion-current-based quantitative platform. More than 2000 protein groups were quantified in all 16 experiments. Two hundred and nine heart-regeneration-related protein groups were quantified and clustered into six time-course patterns. Functional analysis indicated that multiple molecular function and metabolic pathways were involved in heart regeneration. Interestingly, Ingenuity Pathway Analysis revealed that P53 signaling was inhibited during the heart regeneration, which was further verified by real-time quantitative polymerase chain reaction (Q-PCR). In summary, we applied systematic proteomics analysis on regenerating zebrafish heart, uncovered the dynamics of regenerative genes expression and regulatory pathways, and provided invaluable insight into design regenerative-based strategies in human hearts.
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15 MeSH Terms
Optic Nerve Regeneration After Crush Remodels the Injury Site: Molecular Insights From Imaging Mass Spectrometry.
Stark DT, Anderson DMG, Kwong JMK, Patterson NH, Schey KL, Caprioli RM, Caprioli J
(2018) Invest Ophthalmol Vis Sci 59: 212-222
MeSH Terms: Animals, Axons, Cell Count, Cell Survival, Disease Models, Animal, Gliosis, Lipid Metabolism, Male, Microscopy, Confocal, Nerve Crush, Nerve Regeneration, Neuronal Plasticity, Optic Nerve, Optic Nerve Injuries, Rats, Rats, Inbred F344, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Show Abstract · Added March 22, 2018
Purpose - Mammalian central nervous system axons fail to regenerate after injury. Contributing factors include limited intrinsic growth capacity and an inhibitory glial environment. Inflammation-induced optic nerve regeneration (IIR) is thought to boost retinal ganglion cell (RGC) intrinsic growth capacity through progrowth gene expression, but effects on the inhibitory glial environment of the optic nerve are unexplored. To investigate progrowth molecular changes associated with reactive gliosis during IIR, we developed an imaging mass spectrometry (IMS)-based approach that identifies discriminant molecular signals in and around optic nerve crush (ONC) sites.
Methods - ONC was performed in rats, and IIR was established by intravitreal injection of a yeast cell wall preparation. Optic nerves were collected at various postcrush intervals, and longitudinal sections were analyzed with matrix-assisted laser desorption/ionization (MALDI) IMS and data mining. Immunohistochemistry and confocal microscopy were used to compare discriminant molecular features with cellular features of reactive gliosis.
Results - IIR increased the area of the crush site that was occupied by a dense cellular infiltrate and mass spectral features consistent with lysosome-specific lipids. IIR also increased immunohistochemical labeling for microglia and macrophages. IIR enhanced clearance of lipid sulfatide myelin-associated inhibitors of axon growth and accumulation of simple GM3 gangliosides in a spatial distribution consistent with degradation of plasma membrane from degenerated axons.
Conclusions - IIR promotes a robust phagocytic response that improves clearance of myelin and axon debris. This growth-permissive molecular remodeling of the crush injury site extends our current understanding of IIR to include mechanisms extrinsic to the RGC.
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17 MeSH Terms
A novel conduit-based coaptation device for primary nerve repair.
Bamba R, Riley DC, Kelm ND, Cardwell N, Pollins AC, Afshari A, Nguyen L, Dortch RD, Thayer WP
(2018) Int J Neurosci 128: 563-569
MeSH Terms: Animals, Diffusion Tensor Imaging, Disease Models, Animal, Extracellular Matrix, Female, Microsurgery, Nerve Regeneration, Peripheral Nerve Injuries, Rats, Rats, Sprague-Dawley, Sciatic Nerve
Show Abstract · Added October 24, 2018
BACKGROUND - Conduit-based nerve repairs are commonly used for small nerve gaps, whereas primary repair may be performed if there is no tension on nerve endings. We hypothesize that a conduit-based nerve coaptation device will improve nerve repair outcomes by avoiding sutures at the nerve repair site and utilizing the advantages of a conduit-based repair.
METHODS - The left sciatic nerves of female Sprague-Dawley rats were transected and repaired using a novel conduit-based device. The conduit-based device group was compared to a control group of rats that underwent a standard end-to-end microsurgical repair of the sciatic nerve. Animals underwent behavioral assessments at weekly intervals post-operatively using the sciatic functional index (SFI) test. Animals were sacrificed at four weeks to obtain motor axon counts from immunohistochemistry. A sub-group of animals were sacrificed immediately post repair to obtain MRI images.
RESULTS - SFI scores were superior in rats which received conduit-based repairs compared to the control group. Motor axon counts distal to the injury in the device group at four weeks were statistically superior to the control group. MRI tractography was used to demonstrate repair of two nerves using the novel conduit device.
CONCLUSIONS - A conduit-based nerve coaptation device avoids sutures at the nerve repair site and leads to improved outcomes in a rat model. Conduit-based nerve repair devices have the potential to standardize nerve repairs while improving outcomes.
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Immediate Enhancement of Nerve Function Using a Novel Axonal Fusion Device After Neurotmesis.
Riley DC, Boyer RB, Deister CA, Pollins AC, Cardwell NL, Kelm ND, Does MD, Dortch RD, Bamba R, Shack RB, Thayer WP
(2017) Ann Plast Surg 79: 590-599
MeSH Terms: Animals, Axons, Disease Models, Animal, Drug Delivery Systems, Electromyography, Female, Immunohistochemistry, Male, Nerve Regeneration, Neurosurgical Procedures, Peripheral Nerve Injuries, Polyethylene Glycols, Random Allocation, Rats, Rats, Sprague-Dawley, Recovery of Function, Sciatic Nerve, Trauma, Nervous System
Show Abstract · Added October 24, 2018
BACKGROUND - The management of peripheral nerve injuries remains a large challenge for plastic surgeons. With the inability to fuse axonal endings, results after microsurgical nerve repair have been inconsistent. Our current nerve repair strategies rely upon the slow and lengthy process of axonal regeneration (~1 mm/d). Polyethylene glycol (PEG) has been investigated as a potential axonal fusion agent; however, the percentage of axonal fusion has been inconsistent. The purpose of this study was to identify a PEG delivery device to standardize outcomes after attempted axonal fusion with PEG.
MATERIALS AND METHODS - We used a rat sciatic nerve injury model in which we completely transected and repaired the left sciatic nerve to evaluate the efficacy of PEG fusion over a span of 12 weeks. In addition, we evaluated the effectiveness of a delivery device's ability to optimize results after PEG fusion.
RESULTS - We found that PEG rapidly (within minutes) restores axonal continuity as assessed by electrophysiology, fluorescent retrograde tracer, and diffusion tensor imaging. Immunohistochemical analysis shows that motor axon counts are significantly increased at 1 week, 4 weeks, and 12 weeks postoperatively in PEG-treated animals. Furthermore, PEG restored behavioral functions up to 50% compared with animals that received the criterion standard epineurial repair (control animals).
CONCLUSIONS - The ability of PEG to rapidly restore nerve function after neurotmesis could have vast implications on the clinical management of traumatic injuries to peripheral nerves.
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Fabrication of Trabecular Bone-Templated Tissue-Engineered Constructs by 3D Inkjet Printing.
Vanderburgh JP, Fernando SJ, Merkel AR, Sterling JA, Guelcher SA
(2017) Adv Healthc Mater 6:
MeSH Terms: Biocompatible Materials, Bone Regeneration, Cancellous Bone, Cartilage, Cell Differentiation, Cells, Cultured, Humans, Materials Testing, Mesenchymal Stem Cells, Osteogenesis, Printing, Three-Dimensional, Tissue Engineering, Tissue Scaffolds
Show Abstract · Added March 21, 2018
3D printing enables the creation of scaffolds with precisely controlled morphometric properties for multiple tissue types, including musculoskeletal tissues such as cartilage and bone. Computed tomography (CT) imaging has been combined with 3D printing to fabricate anatomically scaled patient-specific scaffolds for bone regeneration. However, anatomically scaled scaffolds typically lack sufficient resolution to recapitulate the <100 micrometer-scale trabecular architecture essential for investigating the cellular response to the morphometric properties of bone. In this study, it is hypothesized that the architecture of trabecular bone regulates osteoblast differentiation and mineralization. To test this hypothesis, human bone-templated 3D constructs are fabricated via a new micro-CT/3D inkjet printing process. It is shown that this process reproducibly fabricates bone-templated constructs that recapitulate the anatomic site-specific morphometric properties of trabecular bone. A significant correlation is observed between the structure model index (a morphometric parameter related to surface curvature) and the degree of mineralization of human mesenchymal stem cells, with more concave surfaces promoting more extensive osteoblast differentiation and mineralization compared to predominately convex surfaces. These findings highlight the significant effects of trabecular architecture on osteoblast function.
© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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13 MeSH Terms
Canonical Wnt Signaling Ameliorates Aging of Intestinal Stem Cells.
Nalapareddy K, Nattamai KJ, Kumar RS, Karns R, Wikenheiser-Brokamp KA, Sampson LL, Mahe MM, Sundaram N, Yacyshyn MB, Yacyshyn B, Helmrath MA, Zheng Y, Geiger H
(2017) Cell Rep 18: 2608-2621
MeSH Terms: Animals, Biomarkers, Cell Count, Cell Proliferation, Cellular Senescence, Female, Intestine, Small, Mice, Organoids, Regeneration, Stem Cell Niche, Stem Cells, Wnt Signaling Pathway
Show Abstract · Added March 19, 2017
Although intestinal homeostasis is maintained by intestinal stem cells (ISCs), regeneration is impaired upon aging. Here, we first uncover changes in intestinal architecture, cell number, and cell composition upon aging. Second, we identify a decline in the regenerative capacity of ISCs upon aging because of a decline in canonical Wnt signaling in ISCs. Changes in expression of Wnts are found in stem cells themselves and in their niche, including Paneth cells and mesenchyme. Third, reactivating canonical Wnt signaling enhances the function of both murine and human ISCs and, thus, ameliorates aging-associated phenotypes of ISCs in an organoid assay. Our data demonstrate a role for impaired Wnt signaling in physiological aging of ISCs and further identify potential therapeutic avenues to improve ISC regenerative potential upon aging.
Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
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13 MeSH Terms
Neurotransmitter-Regulated Regeneration in the Zebrafish Retina.
Rao MB, Didiano D, Patton JG
(2017) Stem Cell Reports 8: 831-842
MeSH Terms: Animals, Cell Proliferation, Neuroglia, Receptors, Glutamate, Regeneration, Retina, Signal Transduction, Stem Cells, Zebrafish, gamma-Aminobutyric Acid
Show Abstract · Added August 4, 2017
Current efforts to repair damaged or diseased mammalian retinas are inefficient and largely incapable of fully restoring vision. Conversely, the zebrafish retina is capable of spontaneous regeneration upon damage using Müller glia (MG)-derived progenitors. Understanding how zebrafish MG initiate regeneration may help develop new treatments that prompt mammalian retinas to regenerate. We show that inhibition of γ-aminobutyric acid (GABA) signaling facilitates initiation of MG proliferation. GABA levels decrease following damage, and MG are positioned to detect decreased ambient levels and undergo dedifferentiation. Using pharmacological and genetic approaches, we demonstrate that GABA receptor inhibition stimulates regeneration in undamaged retinas while activation inhibits regeneration in damaged retinas.
Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
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10 MeSH Terms
The Promise of Cardiac Regeneration by In Situ Lineage Conversion.
Nam YJ, Munshi NV
(2017) Circulation 135: 914-916
MeSH Terms: Animals, Cell Lineage, Cellular Reprogramming, Fibroblasts, Heart, Humans, Induced Pluripotent Stem Cells, Myocytes, Cardiac, Regeneration
Added April 2, 2019
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The challenge of regenerative therapies for the optic nerve in glaucoma.
Calkins DJ, Pekny M, Cooper ML, Benowitz L, Lasker/IRRF Initiative on Astrocytes and Glaucomatous Neurodegeneration Participants
(2017) Exp Eye Res 157: 28-33
MeSH Terms: Animals, Glaucoma, Humans, Nerve Degeneration, Nerve Regeneration, Neuroglia, Optic Disk, Optic Nerve Diseases, Regenerative Medicine, Retinal Ganglion Cells
Show Abstract · Added April 18, 2017
This review arose from a discussion of regenerative therapies to treat optic nerve degeneration in glaucoma at the 2015 Lasker/IRRF Initiative on Astrocytes and Glaucomatous Neurodegeneration. In addition to the authors, participants included Jonathan Crowston, Andrew Huberman, Elaine Johnson, Richard Lu, Hemai Phatnami, Rebecca Sappington, and Don Zack. Glaucoma is a neurodegenerative disease of the optic nerve, and is the leading cause of irreversible blindness worldwide. The disease progresses as sensitivity to intraocular pressure (IOP) is conveyed through the optic nerve head to distal retinal ganglion cell (RGC) projections. Because the nerve and retina are components of the central nervous system (CNS), their intrinsic regenerative capacity is limited. However, recent research in regenerative therapies has resulted in multiple breakthroughs that may unlock the optic nerve's regenerative potential. Increasing levels of Schwann-cell derived trophic factors and reducing potent cell-intrinsic suppressors of regeneration have resulted in axonal regeneration even beyond the optic chiasm. Despite this success, many challenges remain. RGC axons must be able to form new connections with their appropriate targets in central brain regions and these connections must be retinotopically correct. Furthermore, for new axons penetrating the optic projection, oligodendrocyte glia must provide myelination. Additionally, reactive gliosis and inflammation that increase the regenerative capacity must be outweigh pro-apoptotic processes to create an environment within which maximal regeneration can occur.
Copyright © 2017 Elsevier Ltd. All rights reserved.
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10 MeSH Terms
A novel therapy to promote axonal fusion in human digital nerves.
Bamba R, Waitayawinyu T, Nookala R, Riley DC, Boyer RB, Sexton KW, Boonyasirikool C, Niempoog S, Kelm ND, Does MD, Dortch RD, Shack RB, Thayer WP
(2016) J Trauma Acute Care Surg 81: S177-S183
MeSH Terms: Adolescent, Historically Controlled Study, Humans, Lacerations, Male, Nerve Regeneration, Peripheral Nerve Injuries, Peripheral Nerves, Polyethylene Glycols, Recovery of Function
Show Abstract · Added October 24, 2018
BACKGROUND - Peripheral nerve injury can have a devastating impact on our military and veteran population. Current strategies for peripheral nerve repair include techniques such as nerve tubes, nerve grafts, tissue matrices, and nerve growth guides to enhance the number of regenerating axons. Even with such advanced techniques, it takes months to regain function. In animal models, polyethylene glycol (PEG) therapy has shown to improve both physiologic and behavioral outcomes after nerve transection by fusion of a portion of the proximal axons to the distal axon stumps. The objective of this study was to show the efficacy of PEG fusion in humans and to retrospectively compare PEG fusion to standard nerve repair.
METHODS - Patients with traumatic lacerations involving digital nerves were treated with PEG after standard microsurgical neurorrhaphy. Sensory assessment after injury was performed at 1 week, 2 weeks, 1 month, and 2 months using static two-point discrimination and Semmes-Weinstein monofilament testing. The Medical Research Council Classification (MRCC) for Sensory Recovery Scale was used to evaluate the level of injury. The PEG fusion group was compared to patient-matched controls whose data were retrospectively collected.
RESULTS - Four PEG fusions were performed on four nerve transections in two patients. Polyethylene glycol therapy improves functional outcomes and speed of nerve recovery in clinical setting assessed by average MRCC score in week 1 (2.8 vs 1.0, p = 0.03). At 4 weeks, MRCC remained superior in the PEG fusion group (3.8 vs 1.3, p = 0.01). At 8 weeks, there was improvement in both groups with the PEG fusion cohort remaining statistically better (4.0 vs 1.7, p = 0.01).
CONCLUSION - Polyethylene glycol fusion is a novel therapy for peripheral nerve repair with proven effectiveness in animal models. Clinical studies are still in early stages but have had encouraging results. Polyethylene glycol fusion is a potential revolutionary therapy in peripheral nerve repair but needs further investigation.
LEVEL OF EVIDENCE - Therapeutic study, level IV.
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