Three-dimensional visualization of intramuscular innervation in intact adult skeletal muscle by a modified iDISCO method

Li, Yusha; Xu, Jianyi; Zhu, Jingtan; Yu, Tingting; Zhu, Dan

doi:10.1117/1.nph.7.1.015003

Cited by 8 publications

(3 citation statements)

References 32 publications

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“…However, as we showed, their ability to migrate was poor, as it was visualized using iDISCO+ method. This tool was already used to visualize intramuscular innervation in skeletal muscle [ 48 ] and skeletal muscle tissue in teratomas [ 8 ]. In the current study, we show that it can also be applied successfully to analyze the localization of injected cells into muscles, including their potential incorporation into newly formed myofibers.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Muscle Regeneration after BMSC-Conditioned Medium, Syngeneic, or Allogeneic BMSC Injection

Świerczek-Lasek

Tolak

Bijoch

et al. 2022

Cells

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For many years optimal treatment for dysfunctional skeletal muscle characterized, for example, by impaired or limited regeneration, has been searched. Among the crucial factors enabling its development is finding the appropriate source of cells, which could participate in tissue reconstruction or serve as an immunomodulating agent (limiting immune response as well as fibrosis, that is, connective tissue formation), after transplantation to regenerating muscles. MSCs, including those derived from bone marrow, are considered for such applications in terms of their immunomodulatory properties, as their naive myogenic potential is rather limited. Injection of autologous (syngeneic) or allogeneic BMSCs has been or is currently being tested and compared in many potential clinical treatments. In the present study, we verified which approach, that is, the transplantation of either syngeneic or allogeneic BMSCs or the injection of BMSC-conditioned medium, would be the most beneficial for skeletal muscle regeneration. To properly assess the influence of the tested treatments on the inflammation, the experiments were carried out using immunocompetent mice, which allowed us to observe immune response. Combined analysis of muscle histology, immune cell infiltration, and levels of selected chemokines, cytokines, and growth factors important for muscle regeneration, showed that muscle injection with BMSC-conditioned medium is the most beneficial strategy, as it resulted in reduced inflammation and fibrosis development, together with enhanced new fiber formation, which may be related to, i.e., elevated level of IGF-1. In contrast, transplantation of allogeneic BMSCs to injured muscles resulted in a visible increase in the immune response, which hindered regeneration by promoting connective tissue formation. In comparison, syngeneic BMSC injection, although not detrimental to muscle regeneration, did not result in such significant improvement as CM injection.

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Section: Discussionmentioning

confidence: 99%

Comparison of Muscle Regeneration after BMSC-Conditioned Medium, Syngeneic, or Allogeneic BMSC Injection

Świerczek-Lasek

Tolak

Bijoch

et al. 2022

Cells

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“…To properly visualize neuromuscular junctions (NMJ), the traditional iDISCO protocol was modified to an m-iDISCO protocol as previously described. 21 Primary antibodies included beta-3 tubulin (BioLegend) and alpha-bungarotoxin (Fisher) (1:1000) and appropriate secondary antibodies (1:200). Images were taken with an inverted confocal fluorescence microscope (LSM710, Zeiss) equipped with a 5×∕0.25 dry objective (W.D.…”

Section: Immuno-enabled Three-dimensional Imaging Of Surgical Constru...mentioning

confidence: 99%

A Direct Comparison of Targeted Muscle Reinnervation and Regenerative Peripheral Nerve Interfaces to Prevent Neuroma Pain

Senger,

Hardy,

Thorkelsson

et al. 2023

Neurosurgery

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BACKGROUND AND OBJECTIVES: Targeted muscle reinnervation (TMR) and regenerative peripheral nerve interface (RPNI) surgeries manage neuroma pain; however, there remains considerable discord regarding the best treatment strategy. We provide a direct comparison of TMR and RPNI surgery using a rodent model for the treatment of neuroma pain. METHODS: The tibial nerve of 36 Fischer rats was transected and secured to the dermis to promote neuroma formation. Pain was assessed using mechanical stimulation at the neuroma site (direct pain) and von Frey analysis at the footpad (to assess tactile allodynia from collateral innervation). Once painful neuromas were detected 6 weeks later, animals were randomized to experimental groups: (a) TMR to the motor branch to biceps femoris, (b) RPNI with an extensor digitorum longus graft, (c) neuroma excision, and (d) neuroma in situ. The TMR/RPNIs were harvested to confirm muscle reinnervation, and the sensory ganglia and nerves were harvested to assess markers of regeneration, pain, and inflammation. RESULTS: Ten weeks post-TMR/RPNI surgery, animals had decreased pain scores compared with controls (P < .001) and they both demonstrated neuromuscular junction reinnervation. Compared with neuroma controls, immunohistochemistry showed that sensory neuronal cell bodies of TMR and RPNI showed a decrease in regeneration markers phosphorylated cyclic AMP receptor binding protein and activation transcription factor 3 and pain markers transient receptor potential vanilloid 1 and neuropeptide Y (P < .05). The nerve and dorsal root ganglion maintained elevated Iba-1 expression in all cohorts. CONCLUSION: RPNI and TMR improved pain scores after neuroma resection suggesting both may be clinically feasible techniques for improving outcomes for patients with nerve injuries or those undergoing amputation.

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“…5 . 13 This process is therefore required for the restoration of muscle function and the prevention of muscular atrophy, as seen in nervous system conditions such as motor neuron diseases (MND). 128 Motor units, composed of a motor neuron and the group of myofibers it innervates, have been demonstrated to undergo physiological changes due to age-related progressive muscular atrophy.…”

Section: Maturation Of Constructsmentioning

confidence: 99%

Replace and repair: Biomimetic bioprinting for effective muscle engineering

Massey

Boyd‐Moss

et al. 2021

APL Bioengineering

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The debilitating effects of muscle damage, either through ischemic injury or volumetric muscle loss (VML), can have significant impacts on patients, and yet there are few effective treatments. This challenge arises when function is degraded due to significant amounts of skeletal muscle loss, beyond the regenerative ability of endogenous repair mechanisms. Currently available surgical interventions for VML are quite invasive and cannot typically restore function adequately. In response to this, many new bioengineering studies implicate 3D bioprinting as a viable option. Bioprinting for VML repair includes three distinct phases: printing and seeding, growth and maturation, and implantation and application. Although this 3D bioprinting technology has existed for several decades, the advent of more advanced and novel printing techniques has brought us closer to clinical applications. Recent studies have overcome previous limitations in diffusion distance with novel microchannel construct architectures and improved myotubule alignment with highly biomimetic nanostructures. These structures may also enhance angiogenic and nervous ingrowth post-implantation, though further research to improve these parameters has been limited. Inclusion of neural cells has also shown to improve myoblast maturation and development of neuromuscular junctions, bringing us one step closer to functional, implantable skeletal muscle constructs. Given the current state of skeletal muscle 3D bioprinting, the most pressing future avenues of research include furthering our understanding of the physical and biochemical mechanisms of myotube development and expanding our control over macroscopic and microscopic construct structures. Further to this, current investigation needs to be expanded from immunocompromised rodent and murine myoblast models to more clinically applicable human cell lines as we move closer to viable therapeutic implementation.

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Three-dimensional visualization of intramuscular innervation in intact adult skeletal muscle by a modified iDISCO method

Cited by 8 publications

References 32 publications

Comparison of Muscle Regeneration after BMSC-Conditioned Medium, Syngeneic, or Allogeneic BMSC Injection

Comparison of Muscle Regeneration after BMSC-Conditioned Medium, Syngeneic, or Allogeneic BMSC Injection

A Direct Comparison of Targeted Muscle Reinnervation and Regenerative Peripheral Nerve Interfaces to Prevent Neuroma Pain

Replace and repair: Biomimetic bioprinting for effective muscle engineering

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