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> Anatomy > Tissue > Epineurium

Epineurium

Epineurium: The connective tissue sheath surrounding a peripheral nerve trunk or fasciculus.
It is composed of dense irregular connective tissue containing blood vessels that nourish the nerve.
The epineurium provides protection and structural support for the nerve fibers within.
Detailed understanding of the epineurium's anatomy and function is crucial for optimizing peripheral nerve repair and regeneration procedures.

Most cited protocols related to «Epineurium»

1
Intracellular Recordings of Sensory Neurons in Acutely Isolated DRG

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Publication 2012
Action Potentials Animals ARID1A protein, human Bicarbonate, Sodium Cells C Fibers Egtazic Acid Electric Conductivity Enzymes Epineurium Ganglia Glucose HEPES Inflammation Mediators Magnesium Chloride Microelectrodes Microscopy Neuron, Afferent Neurons Nylons Protoplasm Pulses Root, Dorsal Satellite Glia Sodium Chloride Suction Drainage Ventral Roots
2
Sciatic Nerve Dissection and Teasing
The study protocol was approved by the University of Miami Animal Care and Use Committee. Experiments were conducted in accordance with the regulations and guidelines of The University of Miami Animal Care and Use Committee (IACUC), which supervises the work and acts in compliance with all USDA and NIH regulations governing the use of vertebrate animals for experimentation. Adult and neonatal sciatic nerves were obtained from adult female (3-month-old) and postnatal day-2 (P2) Sprague-Dawley rats, respectively. Sciatic nerves were dissected out immediately under aseptic conditions and placed in ice-cold Leibovitz’s L-15 Medium (L-15) medium containing gentamicin.
Each adult sciatic nerve was cleaned off excess tissue including muscle, fat and blood vessels while working under a Stemi 305 Stereo dissecting microscope (Carl Zeiss Microscopy GmbH, Germany) placed inside a horizontal laminar flow hood. After the removal of non-peripheral nerve tissue, each nerve biopsy was weighed and measured. Adult nerve fragments (~2.5 cm in length) weighed 33.9 ± 5.2 mg on average after cleaning. Subsequently, starting at the most proximal end to the spinal cord and with the help of fine forceps, the epineurium was mechanically removed as one single sheath. The epineurium and nerve fibers were individually collected and placed into fresh L-15-containing dishes. The nerve fascicles were extensively teased to the level of individual fibers using Dumont N° 4 and N° 5 fine forceps (Biology tips) with visualization through a dissecting microscope.
The teasing step consisted of repeated pulling of the individual nerve fascicles until fibers of the least possible caliber were obtained. On average, dissection and teasing of one nerve biopsy was completed within 30–45 minutes, depending on the operator’s technical skills. Tissues were transferred regularly to new dishes containing ice-cold L-15 medium for increased safety throughout the process of cleaning, removal of the epineurium and teasing of the fibers. Images of all these steps were acquired using an AxioCam ERc 5s camera coupled to the dissecting microscope and ZEN 2 lite Imaging software 2.0 (Carl Zeizz Microscopy GmbH).
Postnatal nerve fragments were cleaned off non-peripheral nerve tissues essentially as described above with the exception that the tissue was cut into small fragments using fine scissors, rinsed in cold L-15, and immediately subjected to enzymatic dissociation.
Andersen N.D., Srinivas S., Piñero G, & Monje P.V. (2016). A rapid and versatile method for the isolation, purification and cryogenic storage of Schwann cells from adult rodent nerves. Scientific Reports, 6, 31781.
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Publication 2016
Adult Animals Asepsis Biopsy Blood Vessel Common Cold Dissection Enzymes Epineurium Forceps Gentamicin Hyperostosis, Diffuse Idiopathic Skeletal Infant, Newborn Institutional Animal Care and Use Committees Microscopy Muscle Tissue Nerve Fibers Nervousness Neurectomy Peripheral Nerves Rats, Sprague-Dawley Safety Sciatic Nerve SERPINA3 protein, human Spinal Cord ST Segment Elevation Myocardial Infarction Tissues Vertebrates Woman
3
Comprehensive Immunohistochemical Analysis of Nerve Injury
At the described timepoints following surgery, nerves were removed and fixed in 4% paraformaldehyde for 5 hours at 4°C. Following fixation, nerves were then washed in PTX (1% Triton X-100 (Sigma, T9284) in phosphate buffered saline (PBS)) three times for 10 minutes each time. To ensure better antibody penetration for nerve crush samples, the epineurium was removed in these preparations after washing in PTX. Nerves were subsequently incubated with blocking solution (10% foetal bovine serum (FBS) in PTX) overnight at 4°C. The following day, nerves were transferred into primary antibodies in PTX containing 10% FBS and incubated for 48h-72h at 4°C with gentle rocking. Primary antibodies used for the experiments are neurofilament heavy chain (1:1000, Abcam, ab4680), S100β (1:100, DAKO, Z0311), Ki67 (1:100, Abcam, ab15580) and myelin basic protein (1:100, Santa Cruz Biotechnology, sc-13912). After the incubation, nerves were washed with PTX three times for 15 minutes each wash, followed by washing in PTX for 6 hours at room temperature, with a change of PTX every 1 hour. Secondary antibodies (1:500, Invitrogen) and Hoechst dye (1:1000, Invitrogen) were diluted in PTX containing 10% FBS, and incubated with the nerves for 48h at 4°C with rocking. Next, nerves were washed in PTX three times for 15 minutes each, followed by washing in PTX for 6 hours at room temperature, changing the PTX each hour, and then washed overnight without changing PTX at 4°C. For FluoroMyelin Red (Invitrogen, F34652) staining, FluoroMyelin Red in PBS (1:300) was used to stain the nerves for 30 minutes at room temperature. When staining was complete, the nerve was washed 3 times for 10 minutes each in PBS. Nerves were cleared sequentially with 25%, 50%, 75% glycerol (Sigma, G6279) in PBS between 12–24h for each glycerol concentration. Following clearing, nerves were mounted in CitiFluor (Agar Scientific, R1320) for confocal imaging.
Dun X.P, & Parkinson D.B. (2015). Visualizing Peripheral Nerve Regeneration by Whole Mount Staining. PLoS ONE, 10(3), e0119168.
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Publication 2015
Agar Antibodies Epineurium Fetal Bovine Serum Glycerin Immunoglobulins MBP protein, human Nerve Crush Nervousness Neurofilaments Operative Surgical Procedures paraform Phosphates S100 Calcium Binding Protein beta Subunit Saline Solution Stains Triton X-100
4
Realistic Nerve Model for Selective Fascicle Activation
A realistic nerve model was used to illustrate the clinical relevance of our findings. An FEM model of a cadaveric human femoral nerve, based on a histological nerve cross section [24 ], was chosen as an example of a realistic nerve model. A similar model is currently being used to improve the design of nerve cuff electrodes intended for femoral nerve stimulation [17 (link)]. A pair of neighboring fascicles was chosen to demonstrate the thp, Df, and neighboring fascicle effects in a realistic nerve model (Fig. 2). These two fascicles were chosen because of their differences in diameters and functions, which makes selective activation of these fascicles more desired for functional electrical stimulation. Fascicles 1 and 2 had diameters of 200 and 650 μm, respectively. Their centers were 550 μm apart.
Four FEM models were generated: 1), 2) a model of each individual fascicle in the pair, 3) a model of both fascicles in the pair, and 4) a model with all fascicles in the nerve cross section. Unlike our previous models, these models also included epineurium around the fascicles. Four perineurial thicknesses were tested, 0% and 3% of the each fascicle diameter, 30 μm (equal to 15% of fascicle 1 diameter and 4.6% of fascicle 2 diameter), and 50 μm (equal to 25% of fascicle 1 diameter and 7.7% of fascicle 2 diameter). These perineurial thicknesses were chosen based on thickness used in previous models [9 ], [12 (link)], [30 (link)] and our analysis of physiologic perineurial thickness (see Section III). The electrode contact was placed on the inner surface of the cuff and centered between the two fascicles in the chosen pair.
The percent activations of each of the two fascicles alone, with a neighboring fascicle, and with all fascicles in the cross section were compared over a range of recruitment levels (pulse widths of 0.01–0.1 ms). The effects of the Df and thp were also assessed based on the computed percent activations.
Grinberg Y., Schiefer M.A., Tyler D.J, & Gustafson K.J. (2008). Fascicular Perineurium Thickness, Size, and Position Affect Model Predictions of Neural Excitation. IEEE transactions on neural systems and rehabilitation engineering : a publication of the IEEE Engineering in Medicine and Biology Society, 16(6), 572-581.
Publication 2008
Epineurium Homo sapiens Nerves, Femoral Nervousness Perineurium physiology Pulse Rate Stimulations, Electric
5
Sciatic Nerve Crush and Cut Severance Protocol
Prior to surgery, animals were anesthetized using a small mammal isoflurane inhalation system (Handlebar Anesthesia, Austin, TX). Anesthesia was induced initially with a 4% isoflurane/oxygen mixture at a 1.5 L/min flow rate, and then maintained with a 1.5–2% mixture at 1 L/min for the duration of the surgery. The surgical area (lateral aspect of the left hindlimb) was trimmed of fur and disinfected using 10% povidone-iodine. A 2–3 cm long incision was made in the skin above the thigh muscles of the left hindlimb of each rat. The biceps femoris muscle was split parallel to the muscle fibers using dissection scissors to expose the sciatic nerve. The sciatic nerve was trimmed free of any connective tissue using micro-dissection scissors. Crush- or cut-severances were made in either hypotonic Ca2+-free or isotonic Ca2+-containing saline as specified for a given protocol.
We completely crush-severed 1–2 mm lengths of sciatic nerves using Dumont #5 micro-forceps placed perpendicular to the nerve by applying enough force to cause the underlying axons to separate while leaving the epineurium intact. After crush-severance, a small slit was made in the epineurium with micro-scissors to allow solutions to diffuse more readily to axons at the injury site. Crush excision repairs were made by completely excising the crushed nerve segment with dissection scissors. This typically resulted in a 2–3 mm gap between the severed nerve ends, which were then closely re-apposed with micro-sutures similar to our cut-severance protocol (see below). For details of these and other crush-severance surgical procedures, see previous publications (Britt et al., 2010 (link); Bittner et al., 2012 (link)).
Cut-severances were made by transecting the entire sciatic nerve with a single stroke of dissection scissors to completely sever all PNAs as well as their endo-, peri-, and epineural sheaths. Cut axonal ends and sheaths completely separated by 1–3 mm. Bundles of axons would sometimes swell out of the epineural sheath at the cut ends. In some animals as specified, these axonal ends and epineural sheaths were carefully trimmed so that the cut ends formed smooth flat planes that could be closely apposed with minimal gaps or axonal protrusions. Similar axonal trimming was performed, but not always noted, in previous publications (Bittner et al., 2012 (link); Sexton et al., 2012 (link); Rodriguez-Feo et al., 2013 (link); Riley et al., 2015 (link)). Cut-severed nerves were closely re-apposed with 10–0 micro-sutures (Ethicon, Somerville, NJ), leaving enough space between sutures to allow close positioning of a micropipette for diffusion of sterile solutions of PEG, antioxidants, etc., to the lesion site. Unless stated otherwise, cut axonal ends were typically re-apposed within 5–15 min after severance. For additional details of this cut severance surgical procedure, see Bittner et al., (2012) (link).
In this paper unless explicitly stated otherwise, the term PEG-fusion always denotes the following sequence of bio-engineered solutions applied to the lesion site of crush-severed sciatic nerves or cut-severed and micro-sutured sciatic nerves: (1) exposure of cut- or crush-severed axonal ends to hypotonic Ca2+-free saline (PlasmaLyte-A: Baxter, Deerfield, IL), ( (2) application of 1% MB (Acros Organics, Morris Plains, NJ) dissolved in double-distilled water (ddH2O) for 1–2 min, (3) application of 50% by weight PEG (3.35kD; Sigma Aldrich, St. Louis, MO) in sterile ddH2O for 1–2 min, and (4) extensive rinsing with Ca2+-containing isotonic saline (Lactated Ringers: Hospira, Lake Forest, IL). [See Figure 4, Bittner et al., 2015 .] The term “negative control” describes the same procedures and solution applications as above, but without the addition of PEG to a crush- or cut-severed nerve. Note that the ends of cut negative controls are apposed by micro-sutures, the “gold standard” for clinical repair (Isaacs et al., 2010 ; Wolfe et al., 2010 ). As another negative control for dye diffusion experiments, PEG was applied to cut-severed nerves that were not micro-sutured, i.e., cut ends not closely apposed by micro-sutures do not PEG-fuse, but rather PEG seals off the cut ends (“PEG-sealing”. See below and Spaeth et al., 2012b (link)). PEG-fused and negative control animals to be tested for behavioral recovery by SFI tests received 5 mg/kg subcutaneous injection of carprofen after surgery.
Ghergherehchi C.L., Bittner G.D., Hastings R.L., Mikesh M., Riley D.C., Trevino R.C., Schallert T., Thayer W.P., Sunkesula S.R., Ha T.A., Munoz N., Pyarali M., Bansal A., Poon A.D., Mazal A.T., Smith T.A., Wong N.S, & Dunne P.J. (2016). Effects of extracellular calcium and surgical techniques on restoration of axonal continuity by PEG-fusion following complete cut- or crush-severance of rat sciatic nerves. Journal of neuroscience research, 94(3), 231-245.
Publication 2015
Anesthesia Animals Antioxidants austin Axon Biceps Femoris carprofen Cerebrovascular Accident Connective Tissue Diffusion Dissection Endometriosis Epineurium Excision Repair Forceps Forests Gold Hindlimb Inhalation Injuries Isoflurane Lactated Ringer's Solution Lesion of Sciatic Nerve Mammals Muscle Tissue Nerve Crush Nerve Endings Nervousness Operative Surgical Procedures Oxygen Peptide Nucleic Acids Plasmalyte A Povidone Iodine Saline Solution Sciatic Nerve Skin Sterility, Reproductive Subcutaneous Injections Sutures Thigh

Most recents protocols related to «Epineurium»

1
Evaluating Intra-observer Reliability of DTI
A subset of 10 nerve fascicles, interfascicular epineurium areas, perineurial areas, and nerve CSA were randomly selected and segmented again by the same observer 30 days after the primary segmentation to assess intra-observer agreement. Intraclass correlation coefficient (ICC) was calculated from the FA (Koo and Li, 2016 (link)). FA was chosen for ICC calculation as this index is most commonly used DTI readout parameter in clinical environment reflecting the degree of cellular structure alignment (Kronlage et al., 2018 (link)).
Pušnik L., Serša I., Umek N., Cvetko E, & Snoj Ž. (2023). Correlation between diffusion tensor indices and fascicular morphometric parameters of peripheral nerve. Frontiers in Physiology, 14, 1070227.
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Publication 2023
Cellular Structures Epineurium Nervousness Perineurium
2
Quantitative Nerve Microstructure Analysis
The nerve segments of peripheral nerves, acquired in 16 continuous slices of 0.625 mm thickness, were identified on reference T2-weighted images (Figure 1A). Quality assessment of slices was performed, and slices with artifacts and partial volume effect were excluded from further analysis. In each included slice, fascicles, interfascicular epineurium, perineurium, and nerve cross-sectional area (CSA) were segmented. The fascicles were defined as intraneural hypointense oval- or round-shaped tissue circumferentially surrounded by a markedly hyperintense line representing the perineurium. The latter served as a reliable segmentation border (Figure 1B). The perineurium was segmented with a single measurement by two parallel lines, as shown in Figure 1C. The hyperintense tissue between the fascicles was defined as interfascicular epineurium (Figure 1D). The nerve was segmented to include the entire nerve but a minimal proportion of the background (Figure 1E). Segmentations were performed manually with the image processing software ImageJ (National Institutes of Health, Bethesda, Maryland, United States). The area was recorded for each segmentation and expressed as CSA for the nerve and fascicles. The fascicular ratio (FR) was calculated as a net fascicular CSA/nerve CSA, the ratio of perineurium as a net perineurium/nerve CSA, and the ratio of interfascicular epineurium as net interfascicular epineurium/nerve CSA (Tagliafico and Tagliafico, 2014 (link)).
The diffusion tensor was calculated from the acquired three-dimensional data as described previously (Basser et al., 1994 (link); Awais et al., 2022 (link)). For each image voxel, the calculated diffusion tensor was diagonalized, which yielded maps of the tensor eigenvalues D1, D2, and D3 and of the corresponding eigenvectors ( ε⇀1,ε⇀2 , ε⇀3 ). Diffusion tensor and its diagonalization were also calculated for every delineated compartment from the corresponding average diffusion weighted signals of the compartment for 19 different diffusion gradient directions. Regional signal averaging enabled the calculation of the fractional anisotropy (FA), mean diffusivity (MD), and D||/D⊥ using the equations Eqs. 1–3 with less noise for each of the segmented compartments. The calculations were made using the software written in the C programming language, which has been previously developed and specifically modified by the authors (Awais et al., 2022 (link)). MD=D1+D2+D33
FA=32D1−MD2+D2−MD2+D3−MD2D12+D22+D32
D‖/D⊥=D1D2+D32
Pušnik L., Serša I., Umek N., Cvetko E, & Snoj Ž. (2023). Correlation between diffusion tensor indices and fascicular morphometric parameters of peripheral nerve. Frontiers in Physiology, 14, 1070227.
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Publication 2023
Anisotropy Diffusion Epineurium Microtubule-Associated Proteins Nerve Net Nervousness Perineurium Peripheral Nerves Tissues
3
Multivariate Analysis of Nerve Microstructure
Statistical analysis was performed using GraphPad Prism 9 (GraphPad Software Inc., San Diego, United States). The Shapiro-Wilk test was used to evaluate the groups for normality. Because normality and equal variance assumptions were met, the fascicular eigenvalues (D1, D2, and D3), as well as their derived indices (MD, FA, and D||/D⊥), were compared by two-way analysis of variance (ANOVA) followed by Tukey’s posthoc test when appropriate. When comparing DT indices of the perineurium, interfascicular epineurium, and nerve CSA one-way ANOVA followed by Tukey’s posthoc test was employed. The fascicular variability of FA was assessed using the coefficient of variation and then compared between fascicles and within fascicles with two-way ANOVA followed by Tukey’s posthoc test when appropriate. To determine the correlations between the DT indices and parameters at the fascicular level, linear regression was performed for each nerve sample then coefficients were compared using a one-sample t-test. The change of nerve FA in subsequent slices was calculated with linear regression. For the assessment of an intra-observer agreement, one-way ICC was used (Koo and Li, 2016 (link)). Differences were deemed statistically significant at p < 0.05. Data are given as means ± standard deviations, ranges, or percentages when appropriate.
Pušnik L., Serša I., Umek N., Cvetko E, & Snoj Ž. (2023). Correlation between diffusion tensor indices and fascicular morphometric parameters of peripheral nerve. Frontiers in Physiology, 14, 1070227.
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Publication 2023
Epineurium Nervousness Perineurium prisma
4
Sciatic Nerve Regeneration with PRF and Fat Graft
All animals were anesthetized by sevoflurane 250 mL (Sojourn, Piramal) via an inhaler. Following the induction of general anaesthesia, rats were placed in a supine position. First, approximately 1 mL of blood was taken from the rat tail artery and placed in the device for centrifugation at the appropriate speed for the preparation of PRF. The rat was then returned to the prone position. After longitudinal skin incisions, bilateral sciatic nerves were accessed via blunt dissection through the gluteus maximus and biceps femoris muscles. Bilateral sciatic nerves, from the sciatic notch to the bifurcation, were exposed. A 0.5-cm-long epineurium segment was circumferentially excised from the main nerve trunk to initiate scar tissue formation. For the right sciatic nerve, the incision was extended to the popliteal region, and approximately 10×10×0.5 mm3 (link)
 fat grafts were prepared from the adipose tissue in this area. Meanwhile, the fat graft obtained from the popliteal region was mixed with the PRF, whose centrifuge was finished and ready, and it was wrapped around the epineurectomy area. The left nerve segment did not undergo any surgical procedure other than the epineurectomy and was considered the sham (or control) group. The right sciatic nerve was considered the experimental (or treatment) group. For histopathological examinations, 12 randomly selected rats were euthanized in the fourth week for early results and the remaining 12 rats in the eighth week for late results.
Kastamoni M., Yavaş S.E., Ozgenel G.Y, & Ersoy S. (2023). The effects of fat graft and platelet-rich fibrin combination after epineurectomy in rats. Revista da Associação Médica Brasileira, 69(2), 272-278.
Publication 2023
Animals Arteries Biceps Femoris BLOOD Buttocks Centrifugation Cicatrix Dissection Epineurium General Anesthesia Grafts Inhaler Medical Devices Nervousness Operative Surgical Procedures Physical Examination Rattus norvegicus Sciatic Nerve Sevoflurane Skin Tail Tissues
5
Quantifying Macrophage and Schwann Cell Recruitment in Peripheral Nerve Regeneration
S100β eGFP mice were euthanized at 10 days after repair (for both treatment groups) to determine macrophage recruitment and SC migration. The regenerative bridge was harvested within the conduit by transecting the proximal and distal sciatic nerve stumps 1 mm from the end of the 5-mm conduit. The epineurial sutures were cut and the regenerative bridge was removed from the conduit and fixed overnight in 4% paraformaldehyde. To stain macrophages, the bridge was then incubated in goat-blocking serum and casein mixture for 5 h. The sample was washed in PBS and then incubated in macrophage-specific primary antibody (1:50 CD68, Abcam) overnight. After the wash step, the regenerative bridge was transferred into secondary antibody solution (1:200 biotinylated goat anti-rabbit IgG, Vector Laboratories) and incubated overnight. Sample was washed again in PBS and transferred into Texas-Red conjugate solution (1:200, Life Technologies) and incubated overnight. The sample was then washed and incubated in DAPI solution (1:1000) overnight. After the final wash in PBS, the sample was immersed into DIX solution (a mixture of n-methyl-d-glucamine, diatrizoic acid, 60% iodixanol and 2% sodium azide) to clear the tissue. All the wash steps were done twice in PBS over a rocking table for 30 min each. All the incubation steps were done at room temperature on a turning wheel at 20–40 rpm.
Samples were then mounted on slides while immersed in DIX solution and the entire regenerative bridge was imaged using tiled, Z stack imaging (Zeiss 710 confocal). The macrophage and SC presence and distribution within the regenerative bridge was quantified by the summation of the fluorescent intensity of each channel (GFP green: SC, Texas red: macrophage, DAPI blue: all cells) at 250 µm intervals from the distal end of the proximal stump using ImageJ. Results were analyzed with group—agarose only (control) or agarose with IL10 (15 µg/mL) concealed to the observer. Differences in total numbers of macrophages were determined using two-tailed t test with P < 0.05.
Golshadi M., Claffey E.F., Grenier J.K., Miller A., Willand M., Edwards M.G., Moore T.P., Sledziona M., Gordon T., Borschel G.H, & Cheetham J. (2023). Delay modulates the immune response to nerve repair. NPJ Regenerative Medicine, 8, 12.
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Publication 2023
Amputation Stumps anti-IgG Caseins Cells Cloning Vectors DAPI Diatrizoic Acid Epineurium Goat IL10 protein, human Immunoglobulins iodixanol Macrophage Mus paraform Rabbits Regeneration S100 Calcium Binding Protein beta Subunit Sciatic Nerve Sepharose Serum Sodium Azide Stains Sutures Tissues

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More about "Epineurium"

Epineurium is a crucial component of the peripheral nervous system, acting as a protective sheath surrounding nerve trunks and fasciculi.
This dense irregular connective tissue contains blood vessels that nourish the nerve fibers within, providing both structural support and a barrier against external factors.
Understanding the detailed anatomy and function of the epineurium is vital for optimizing peripheral nerve repair and regeneration procedures.
Key related terms include the endoneurium (the connective tissue sheath surrounding individual nerve fibers) and the perineurium (the connective tissue sheath surrounding nerve fasciculi).
Researchers studying the epineurium often utilize a range of techniques and materials, such as Forskolin (a compound that can promote nerve regeneration), Collagenase and Trypsin (enzymes used for tissue dissociation), Sprague-Dawley rats (a common animal model), Fetal calf serum (a nutrient-rich medium component), Qiazol (a reagent for RNA extraction), Collagenase I (an enzyme for extracellular matrix digestion), Penicillin and Streptomycin (antimicrobial agents), and DMEM/F12 (a cell culture medium).
These tools and resources can help researchers gain a deeper understanding of epineurium structure, function, and its role in peripheral nerve repair and regeneration.
By leveraging the insights and techniques described here, researchers can enhance the reproducibility and accuracy of their epineurium studies, unlocking new discoveries and optimizing their research outcomes.