A model for ex vivo spinal cord segment culture—A tool for analysis of injury repair strategies

Zhang, Jie; O’Carroll, Simon J.; Wu, Ann Chen; Nicholson, Louise; Green, Colin

doi:10.1016/j.jneumeth.2010.07.012

Cited by 14 publications

(11 citation statements)

References 42 publications

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“…This particular antisense sequence has been used in animal skin wound, spinal cord, and optic nerve models with vehicle or sense controls used. 18,19,[21][22][23] In this study, antisense treatment has been compared with standard of care. For immunohistochemical studies, the following primary antibodies were used: monoclonal mouse anti-Cx43 primary antibody (a gift from David Becker 24 ), monoclonal mouse anti-␣-SMA antibody (Novocastra Laboratories Ltd, Newcastle upon Tyne, UK) and rabbit polyclonal anti-laminin-1 antibody (Sigma Chemical, St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

Improved Corneal Wound Healing through Modulation of Gap Junction Communication Using Connexin43-Specific Antisense Oligodeoxynucleotides

Grupcheva

Laux

Rupenthal

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Gap junctions play a major role in corneal wound healing. This study used reproducible models of corneal wound healing to evaluate the effect of a gap junction channel modulator, connexin43 (Cx43) antisense oligodeoxynucleotides (AsODN), on corneal healing dynamics. METHODS. A mechanical scrape wound model was used to evaluate Cx43 AsODN penetration and initial wound reepithelialization 12 hours postsurgery. Thereafter, detailed analyses of corneal edema, inflammation, and healing were performed in an excimer laser surface ablation model. In vivo confocal microscopy determined clinical parameters (edema, haze) and cellular changes (stromal hypercellularity, reepithelialization), whereas histology and immunohistochemistry were used to quantify stromal edema, inflammation, and reepithelialization. RESULTS. Cx43 AsODN penetrated through the hydrophilic stroma where the epithelium had been removed and accumulated in the basal epithelium close to the wound edge. Twelve hours after scrape wounding, Cx43 AsODN-treated eyes showed a significant reduction in wound area compared with the vehicle alone (1.59 Ϯ 0.37 and 2.29 Ϯ 0.58 mm 2 , respectively, P Ͻ 0.01). After excimer laser ablation, stromal edema and inflammation were reduced, with endothelial structures being clearly visible, and reepithelialization rates were again increased in Cx43 AsODN-treated eyes. Histologic analysis confirmed reduced edema in the central wound site and at the periphery of treated corneas (P Ͻ 0.05), whereas immunohistochemistry showed lower Cx43 levels (P Ͻ 0.05), reduced myofibroblast activation, and improved epithelial basal lamina deposition in antisense-treated wounds (P Ͻ 0.01). CONCLUSIONS. Application of Cx43 AsODN to the cornea reduces stromal edema and inflammation, promoting faster wound closure and a more uniform repair of the epithelial basal lamina after laser ablation. (Invest Ophthalmol Vis Sci.

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Section: Methodsmentioning

confidence: 99%

Improved Corneal Wound Healing through Modulation of Gap Junction Communication Using Connexin43-Specific Antisense Oligodeoxynucleotides

Grupcheva

Laux

Rupenthal

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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“…This drug class can have several benefi cial effects such as reducing heart rate, contractility of the myocardium and the blood pressure, limiting cardiac remodeling, and decreasing the risk for another MI [ 85 ]. β1-Selective antagonists are routinely used in the treatment of HF [ 102 ]; however, in the COMET study it was demonstrated that carvedilol, a nonselective agent which blocks both β1and β2-adrenoceptors, is more benefi cial compared to β1-selective agents (metoprolol) [ 103 ]. Cardiomyocytes express predominantly the β1-adrenoceptor, while the β2-adrenoceptor is the receptor type mainly expressed in cardiac fi broblasts [ 104 ].…”

Section: Mineralocorticoid-receptor Antagonistsmentioning

confidence: 99%

“…In addition the Pluronic gel is also thought to act as a mild surfactant and aid the direct entry of the AsODN into the cells. This Cx43 AsODN 30mer has been used in several experimental disease models including brain slice [98,99], optic nerve ischemia [100], skin wound healing [23,54,101], skin burns [102] and spinal cord injury [20,103]. This AsODN has demonstrated anti-inflammatory and wound healing effects that will be described in detail in the following sections.…”

Section: Non Specific Gap Junction Blockersmentioning

confidence: 99%

“…Spinal cord segments excised from p7 rat pups and placed into organotypic culture show edema, astrogliosis and neuronal cell death [19]. When these segments are treated with either Cx43 specific AsODN [103] or Cx43 mimetic peptides [19] within the first hours after removal from the animal, there is a reduction in connexin43 levels, reduced astrocytosis (GFAP levels) and reduced loss of NeuN and SMI-32 positive neurons in a concentration and time dependent manner [19]. This ex vivo model has also been used to investigate the potential of Cx43 regulation as a tool for testing spinal cord injury repair strategies [103] and this will be discussed in more detail in section 4.2.…”

Section: Spinal Cord Injurymentioning

confidence: 99%

“…We have developed an ex vivo model of spinal cord injury to test intervention strategies model using Cx43 AsODN [103]. In this model Cx43 AsODN application reduces lesion spread and inflammation.…”

Section: Adult Hippocampal Slice Culturementioning

confidence: 99%

See 2 more Smart Citations

Wound Regeneration and Repair

Gourdie¹,

Myers²

2013

Methods in Molecular Biology

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Effects of the combination therapy of electric field stimulation and polyethylene glycol in the ex vivo spinal cord of female rats after compression

Zhang

Wang

Zhang

et al. 2021

J of Neuroscience Research

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The application of electric field stimulation (EFS) can reduce the cation influx after spinal cord injury. However, regenerated cation influx and reestablished injury potential are observed after EFS. Polyethylene glycol (PEG) is popular as an effective cell membrane fusion agent. This study aims to determine the effects of the combination therapy of EFS and PEG in the ex vivo spinal cord after compression. The ex vivo spinal cords of female rats with compression injury were incubated in a double sucrose gap recording chamber (DSGRC) and randomly divided into the following four groups: (1) compression group: compression only, (2) EFS group: EFS for 15 min, (3) PEG group: PEG treatment for 4 min, and (4) EFS + PEG group: EFS for 15 min and PEG treatment for 4 min. The hematoxylin–eosin staining was performed to measure the necrotic area of the spinal cords. The gap potential was detected, and the area under the curve of the gap potential was calculated. The intracellular cation concentration, membrane permeability, and compound action potential were measured and quantified. Results revealed no significant difference in the necrotic areas among different groups, and the compression model of the ex vivo spinal cord in the DSGRC had high consistency and stability. The combination therapy could attenuate cation inflow, promote cell membrane restoration, and promote the functional recovery of the spinal cord conduction after compression in ex vivo spinal cords.

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A model for ex vivo spinal cord segment culture—A tool for analysis of injury repair strategies

Cited by 14 publications

References 42 publications

Improved Corneal Wound Healing through Modulation of Gap Junction Communication Using Connexin43-Specific Antisense Oligodeoxynucleotides

Improved Corneal Wound Healing through Modulation of Gap Junction Communication Using Connexin43-Specific Antisense Oligodeoxynucleotides

Wound Regeneration and Repair

Effects of the combination therapy of electric field stimulation and polyethylene glycol in the ex vivo spinal cord of female rats after compression

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