• Archives of Reconstructive Microsurgery
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• 제6권1호
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• pp.9-28
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• 1997

Adequate vascularization is pivotally essential for a successful nerve graft. Theoretically, the immediate vascularization will inhibit fibroblast infiltration and stimulate nerve cell regeneration. In this study, histomorphological and electrophysiological studies were performed to determine if vascularized grafts are functionally superior. In rat model, a 4cm segment of the sciatic nerve was obtained and placed as a non vascularized graft on one side, and as a vascularized graft connected to the inferior gluteal vessels on the opposite side. To determine the compound action potential of the gastrocnemius muscle, electromyography was done after 2, 3 and 4 months. Histomorphologically, the distribution of myelinated nerve fibers and Schwann cell were evaluated after toluidine blue staining, The following resutls were obtained: 1. The electrophysiological studies showed no difference between the nonvascularized and vascularized grafts. 2. Two and three months after grafting, myelinated nerve fibers were more abundant in the vascularized proximal, middle and distal areas in all nerve fibers of varying diameters. 3. In the post-nonvascularized graft 2-month group, a few myelinated nerve fibers were present in the proximal and middle areas, but none distally. In the post-vascularized graft 2 month group, myelinated nerve fibers ranging $2-8{\mu}m$ were present in all three areas. 4. In the post-nonvascularized graft 3 month group, a few myelinated nerve fibers ranging in $2-6{\mu}m$ were present in all three areas, but in the post-vascularized graft 3 month group, many myelinated nerve fibers ranging in $2-10{\mu}m$ were present in all three areas. 5. In the post-graft 4-month group, more myelinated nerve fibers were present in all three areas of the vascularized grafts. However, nerve fibers of less than $2{\mu}m$ in diameter were more abundant in the non vascularized grafts. 6. Schwann cells were more abundant in the proximal, middle and distal areas of the post-vascularized 2, 3 and 4-month grafts. Based on these findings, the immediate restoration of circulation in vascularized nerve grafts allows for the increased number of surviving Schwann cells, rapid healing of the axon and myelin sheath changes which occur during Wallerian degeneration, and thus is able to stimulate a morphologically optimal regeneration.

• Archives of Reconstructive Microsurgery
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• 제6권1호
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• pp.29-38
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• 1997

A peripheral nerve when approximation of the ends imparts tension at the anastomosis and with a relatively long segment defect after excision of neuroma and neurofibroma cannnot be repaired by early primary suture. The one of the optimistic reconstruction method of severed peripheral nerves is to restore tension-free continuity at the repair site putting an autogenous nerve graft into the neural gap despite of ancipating motor or sensory deficit of the donor nerve area. To overcome the deficit of the autogenous nerve graft, several other conduits supplying a metabolically active environment which is able to support axon regeneration and progression, providing protection against scar invasion, and guiding the regrowing axons to the distal stump of the nerve have been studied. An author have used ipsilateral femoral vein, ipsilateral femoral vein filled with fresh thigh muscle, and autogenous sciatic nerve for the sciatic nerve defect of around 10 mm in length to observe the regeneration pattern in rat by light and electron microscopy. The results were as follows. 1. Light microscopically regeneration pattern of nerve fibers in the autogenous graft group was more abundant than vein graft and vein filled with muscle group. 2. On ultrastructural findings, the proxial end of the graft in various groups showed similar regenerating features of the axons, myelin sheaths, and Schwann cells. The fascicular arrangement of the myelinated and unmyelinated fibers was same regardless of the type of conduits. There were more or less increasing tendency in the number and the diameter of myelinated fibers correlated with the regeneration time. 3. In the middle of the graft, myelinated nerve fibers of vein filled with muscle group were more in number and myelin sheath was thinner than in the venous graft, but the number of regenerating axons in autogenous nerve graft was superior to that in both groups of the graft. The amount of collagen fibrils and amorphous materials in the endoneurial space was increased to elapsed time. 4. There was no difference in regenerating patterns of the nerve fibers of distal end of the graft. The size and shape of the myelinated nerve fbers were more different than that of proximal and middle portion of the graft. From the above results, the degree of myelination and regenerating activity in autogenous nerve is more effective and active in other types of the graft and there were no morphological differences in either ends of the graft regardless of regeneration time.

• 대한수의학회지
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• 제9권1호
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• pp.1-9
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• 1969

The experimental studies were performed to observe the characteristics of the myelinated nerve fibers of the tradiaphragmatic branches of phrenic nerves in dog In the work to be reported five mixed breed Korean dog were used, they were one year old and he body eights were about 12 Kgm. in average with healthy conditions. The specimens (nerve) were taken at the point of 1.5 cm posterior part from the left and right phrenic nerves entrance to the diaphragm, and the dorso-lateral and dorso-medial branches of phrenic nerves vere also, taken at the point of 0.5 cm distal part from their branching portion. The specimens were fixed for 24 hrs. in Flemming's solution, and embedded in paraffin. The paraffin section. were cut at 6 microns and stained with Walter's modification of Weigert-pal method for the myelinated fibers. Microphotographs were taken and enlarged into 750 times of the actual size, and the diameter of the photographic images of the myelinated fibers were measured by the scale on the transparent Percepex Plate. The following conclusions were made: 1. The percentage of distribution of the several intradiaphragmatic branches of phrenic nerves were as follows. Ventral branches were 34.97%, lateral branches were 37.90%, dorsal branches were 27.13%, and the dorsal branches were branched again into dorso-lateral branches(54.22%) and dorso-medial branches (45. 78%). 2. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left ventral branches were $490.80{\pm}12$, and at the right ventral branches were $486.6{\pm}13$. Total average cross sectional area on the left ventral branches were 38,000. $6{\pm}136{\mu}^2$, and the right ventral branches were $37,150.2{\pm}1.782{\mu}^2$. 3. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left lateral branches were $533.8{\pm}8$, and at the right lateral branches were $525.6{\pm}7$. Total average cross sectional area of the left lateral branches were $41,582{\pm}1,170{\mu}^2$, and the right lateral branches were $40,454.8{\pm}812{\mu}^2$. 4. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left dorsal branches were $378.2{\pm}14$, and at the right dorsal branches were $380.2{\pm}8$. Total average cross sectional area of the left dorsal branches were $27,771{\pm}1,256{\mu}^2$, and the right dorsal branches were $27,507.2{\pm}645{\mu}^2$. 5. The mean value and S.D. of the total numbers of the myelinated fibers at the left dorso-lateral branches were $205.4{\pm}15$, and at the right dorso-lateral branches were $210.6{\pm}17$. Total average cross sectional area of the left dorso lateral branches were $15,354. 8{\pm}1,519{\mu}^2$, and the right dorsal branches were $15,887{\pm}1,297{\mu}^2$. 6. The mean value and S.D. of the total numbers of the myelinated nerve fibers at the left dorso-medial branches were $175{\pm}14$, and at the right dorso-medial branches were $176.2{\pm}17$. Total average cross sectional area of the left dorso-medial branches were $13,037.4{\pm}944{\mu}^2$, and at the right dorso-medial branches were, $13,103{\pm}1,373{\mu}^2$. 7. The highest frequent distribution of the intradiaphragmatic branches of phrenic nerves were found in the $10-12{\mu}^2$ groups, and they were almost 30% of the total groups. 8. The largest fibers diameter were in the $14-16{\mu}^2$ groups, and these were shown the lowest frequent distribution. 9. The largest cross sectional area of the intradiaphragmatic branches of phrenic nerves were found in the $10-12{\mu}^2$ groups. 10. All of the intradiaphragmatic branches of phrenic nerves were unimodal which has a peak in the $10-12{\mu}^2$ groups.

• Journal of Korean Biological Nursing Science
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• 제8권1호
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• pp.39-48
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• 2006

Purpose: To see the effects of capsaicin on the peripheral nerve damage of intervibrissal fur in mature rats, Method: 24 female mature rats($200{\sim}250g$) are divided to 3 groups and compared with each other. Immunofluorescence dye using CGRP and PGP antibodies was performed and 8 weeks after administration of capsaicin with control group. Result: The immunopositive reaction against PGP and CGRP was reduced by the damage of epidermal and dermal endings in unmyelinated sheath and thin myelinated sheath and the group after 8weeks showed distinct positive reaction of PGP and CGRP than the group after 4 weeks which means the recover of nerves. Conclusion: As a result, capsaicin influenced on pain-related neurotransmitter like CGRP when administerd to mature rats and even though it caused the damages on unmyelinated sheath and thin myelinated sheath, the damaged nerves recovered after 8 weeks. Also the research about sensory nerve endings scattered over middle dermal and deep epidermal layers such as lanceolate, merkel reticular, Ruffini endings should be studied when the research of the inner conical body is performed. Further studies are necessary about the toxicity and effect of capsaicin on the peripheral nerve endings.

• 한국항행학회논문지
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• 제14권6호
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• pp.1002-1009
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• 2010

본 논문에서는 축삭의 전도 현상을 분석하기 위해서 축삭을 전기전도 모델로 만든 후 여기에 키르히호프의 전류법칙과 전압법칙을 적용한 등가회로를 구성하였다. 축삭의 미세거리 변화에 의한 전위의 거동을 분석하기 위하여 축삭의 각종 파라미터를 구하였다. 검토결과에 의하면 무수신경인 경우 직경을 미지수로 하여 인체의 파라미터를 이용하면 그 속도는 직경의 제곱근에 비례한다. 한편 유수신경인 경우는 그 속도가 직경에 직접적으로 비례한다. 미엘린이 없는 막과는 달리 미엘린껍질의 전도는 전압과 무관하므로 이러한 막 양단의 Hodgkin-Huxley 형 전도모델이 매우 정교하지는 않다.

• Applied Microscopy
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• 제36권2호
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• pp.131-140
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• 2006

Neurofascin은 L1CAM의 하나로 신경섬유의 발달과정에서 중요한 역할을 하는 것으로 알려져 있다. 말초신경의 수초형성과 관련된 neurofascin의 역할을 알아볼 목적으로 면역형광염색과 면역전자현미경기법을 이용하여 랫드의 수초좌골신경섬유에서 neurofascin의 분포를 추구하여 다음과 같은 결과를 얻었다. 1.수초형성이 진행됨에 따라 좌골신경섬유에서 neurofascin 분포는 매우 심하게 변화되었다. 2. 수초신경섬유에서 neurofascin은 Ranvier마디에서 약하게 국재하였다. 3. Neurofascin은 수초신경섬유의 paranodal loop, Schmidt-Lantermann incisure, 속축삭사이막, 바깥축삭사이막처럼 Schwann세포의 막이 밀착되지 않은 부위에서도 뚜렷하게 국재하였다. 이상의 연구결과로 neurofascin은 Schwann세포의 마주보는 막사이에 이상적인 간격을 유지하는데 어떤 역할을 하는 것으로 생각되며, 수초층에서 물질이동이 가능하게 하는 것으로 보인다.

• 한국임상수의학회지
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• 제19권3호
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• pp.316-321
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• 2002

The purpose of this study was to examine the effect of Ga-As(Gallium-Arsenide, wave length; 904 nm) infrared laser irradiation on healing of the experimentally crush injured rat sciatic nerves. The bilateral sciatic nerves of 43 adult male Sprague-Dawley rats were compressed surgically with a straight hemostat (1 mm width). The right legs of all the rats were irradiated using a 27 mW Ga-As infrared laser (laser irradiated group). The radiation procedure was administered for 3 minutes every day for 1, 3, 5, and 7 weeks in each group. Left legs were not irradiated and served as the control group. The numbers of total myelinated axon and degenerated myelin in the sciatic nerves of bilateral legs were measured and analyzed with mage analysis system in order to make a morphological analysis of the effect of the Ga-As infrared laser on injured nerves. Total number of myelinated axons was increased with time interval, especially in the 1, 3. and 5 week of irradiated group. Conversely, the number of degenerated myelin was decreased with time interval, especially in the irradiated group. The effects in the irradiated group were more pronounced than those of the control group. In conclusion, the Ga-As infrared laser irradiation is a useful adjuvant therapy to the regeneration of the peripheral nerve injury.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권5호
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• pp.360-365
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• 2010

Introduction: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. Immunohistochemical staining for neurofilament protein 200 kDa (NFP200) was performed to identify the demyelinated but parent myelinated axons. Materials and Methods: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. Results: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. In the radicular pulp, unmyelinated axons and myelinated axons were present together. However, in the coronal pulp, unmyelinated axons were most common and NFP200 immuno-positive unmyelinated axons with a larger diameter than those in the radicular pulp were observed more frequently. On the other hand, most of the immuno-positive unmyelinated fibers were similar in size to that of typically well-known unmyelinated fibers. Conclusion: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. After the loss of myelin, the size of the axoplasm also decreased.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권1호
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• pp.1-16
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• 2004

The use of artificial nerve conduit containing viable Schwann cells is one of the most promising strategies to repair the peripheral nerve injury. To fabricate an effective nerve conduit whose microstructure and internal environment are more favorable in the nerve regeneration than existing ones, a new three-dimensional Schwann cell culture technique using $Matrigel^{(R)}$. and dorsal root ganglion (DRG) was developed. Nerve conduit of three-dimensionally arranged Schwann cells was fabricated using direct seeding of freshly harvested DRG into a $Matrigel^{(R)}$ filled silicone tube (I.D. 1.98 mm, 14 mm length) and in vitro rafting culture for 2 weeks. The nerve regeneration efficacy of three-dimensionally cultured Schwann cell conduit (3D conduit group, n=6) was assessed using SD rat sciatic nerve defect of 10 mm, and compared with that of silicone conduit filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method (2D conduit group, n=6). After 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were examined using image analyzer and electromicroscopic methods. The SFI and ankle stance angle (ASA) in the functional evaluation were $-60.1{\pm}13.9$, $37.9^{\circ}{\pm}5.4^{\circ}$ in 3D conduit group (n=5) and $-87.0{\pm}12.9$, $32.2^{\circ}{\pm}4.8^{\circ}$ in 2D conduit group (n=4), respectively. And the myelinated axon was $44.91%{\pm}0.13%$ in 3D conduit group and $13.05%{\pm}1.95%$ in 2D conduit group to the sham group. In the TEM study, 3D conduit group showed more abundant myelinated nerve fibers with well organized and thickened extracellular collagen than 2D conduit group, and gastrocnemius muscle and biceps femoris tendon in 3D conduit group were less atrophied and showed decreased fibrosis with less fatty infiltration than 2D conduit group. In conclusion, new three-dimensional Schwann cell culture technique was established, and nerve conduit fabricated using this technique showed much improved nerve regeneration capacity than the silicone tube filled with $Matrigel^{(R)}$ and Schwann cells prepared from the conventional plain culture method.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.295-307
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• 2006

Styela clava, called non-native tunicate or sea squirt, is habitat which include bays and harbors in Korea and several sites in the sea faced world. We fabricate cellulose membrane nerve conduit (CMNC) from this native sea squirt skin, and evaluate the capacity of promoting peripheral nerve regeneration in the rat sciatic nerve defect model. After processing the pure cellulose membrane from the sea squirt skin as we already published before, CMNC was designed as a non-tubular sheet with 14 mm length and 4 mm width. Total eleven male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into sham group (n=2), silicone tube grafted control group (n=3) and experimental group (n=6). Each CMNC grafted nerve was evaluated after 4, 8 and 12 weeks in the experimental group, and after 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were all examined using image analyzer and electromicroscopic methods in the all groups. The regenerated axon and nerve sheath were found only in the inner surface of the CMNC after 4 weeks and became more thicker after 8 and 12 weeks. In the TEM study, CMNC grafted group showed more abundant organized myelinated nerve fibers with thickened extracellular matrix than silicone conduit grafted group after 12 weeks. The sciatic function index (SFI) and ankle stance angle (ASA) in the functional evaluation were $-47.2{\pm}3.9$, $35.5^{\circ}{\pm}4.9^{\circ}$ in CMNC grafted group (n=2) and $-80.4{\pm}7.4$, $29.2^{\circ}{\pm}5.3^{\circ}$ in silicone conduit grafted group (n=3), respectively. And the myelinated axon was 41.59% in CMNC group and 9.51% in silicone conduit group to the sham group. The development of a bioactive CMNC to replace autogenous nerve grafts offers a potential and available approach to improved peripheral nerve regeneration. As we already published before, small peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx, induced the effective axonal regeneration with rapid growth of Schwann cells beneath the inner surface of CMNC. So the possibilities of clinical application as a peripheral nerve regeneration will be able to be suggested.