홍화(Carthamus tinctorius L.)씨 분말의 랫드 골절에 대한 치유 효과

박창현,엄창섭,배춘식,Park, Chang-Hyun,Uhm, Chang-Sub,Bae, Chun-Sik 한국현미경학회 2001 Applied microscopy Vol.31 No.4

12주령 랫드의 비골의 골절을 유발한 후 골절치유에 미치는 홍화씨 분말의 영향을 알아보고자 실험을 실시한 결과 다음과 같은 결과를 얻었다. 비골의 골절은 골절유발 후 5주에 성숙된 신생골 조직으로 충만되어 조직형태학적으로 완전한 유합이 이루어지는데, 홍화씨 분말을 투여한 결과 골절유발 후 4주에 성숙된 신생골 조직으로 골절단이 충만되어 완전한 유합을 이룬 후 5주에서는 골수강도 개통되는 등 대조군과 비교하여 유합시기를 기준으로 1.5주 정도의 빠른 골절의 치유가 이루어졌다. 이상의 결과는 홍화씨 분말에 골절의 치유를 촉진시키는 성분이 포함되어 있을 가능성을 시사한다고 사료된다. Safflower (Carthamus tinctorius L.) is a thistle-like annual plant mainly grown in dry hot climates as an oilseed or birdseed. Traditionally, the oil has mainly been sold in the health food market because it is unsaturated having high linoleic and oleic acid levels. With increased health consciousness in recent years, the oil quality has become a more general health issue. This study was designed to understand whether safflower seed powder has positive effects on the fracture healing in rats. Simple transverse fracture of rat fibula was made with a rotating diamond disc saw. The histologic changes of rats were observed with a scanning electron microscope. The fractured fibulae showed a complete fusion at the fracture site in the 4th to 5th week after a simple transverse fracture. Administration of safflower seed powder facilitated the speed of histologic changes without affecting qualitative changes. These results suggest that safflower seed powder nay have substances that help the fracture healing process.

t-Butyl Alcohol 동결건조법을 이용한 흰쥐 간장의 주사전자현미경적 관찰

엄창섭,박은경,박창현,Uhm, Chang-Sub,Park, Eun-Kyung,Park, Chang-Hyun 한국현미경학회 1998 Applied microscopy Vol.28 No.3

T-butyl alcohol (TBA) freeze-drying method originally designed by Inoue and Osadake (1989) was adopted to dry specimens for scanning electron microscopy and the results were compared with those dried using critical point dryer (CPD). Small pieces $(1\times1\times3mm)$ of liver of Sprague-Dawley rats were cut and fixed in 2% glutaraldehyde in 0.1 M sodium cacodylate buffer after anesthesia, and processed for scanning electron microscopy by several modifications of TBA freeze-drying methods and by the standard preparation method using CPD. The bile canaliculi and sinusoidal endothelial surface were observed. Tissue dehydrated with TBA before TBA freeze-drying preserved the structures best comparable to those prepared with CPD. This result suggests that combination of dehydration with TBA and TBA freeze-drying is a superior method to the original TBA freeze-drying method dehydrated with ethanol.

동시배양한 흰쥐 척수앞뿔신경세포-근육세포 접촉부위에서의 신경결합물질의 분포

엄창섭(Chang-Sub Uhm),서영석(Young-Suk Suh) 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.4

근육신경연접의 형성과정은 세포생물학 및 신경과학분야에서 세계적으로 많은 연구가 진행되고 있는 과제이며, 이 과정 중에 세포 사이의 물리적 접촉 혹은 유착이 중요하게 작용할 것으로 추정되고 있다. 그러나, 세포첩촉물질 특히 신경세포유착물질이 어떻게 변화되는지는 아직 규명되지 않은 부분이다. 본 연구에서는 근육신경연접을 형성하는데 관여하는 척수앞뿔신경세포와 뼈대근육세포를 Sprague-Dawley계 흰쥐에서 분리하여 동시에 배양하고 이 세포들 사이의 접촉 부위, 특히 신경돌기와 근육세포의 접촉부위를 투과전자현미경으로 관찰하고, 몇 가지 세포결합물질의 발현을 면역조직화학염색을 통하여 살펴보았다. 그 결과 척수앞뿔신경세포의 돌기와 뼈대근육세포는 아주 밀접하게 접촉하고 있으며, 신경연접소포를 가지고 있어 신경 근육연접으로 분화될 부위임을 확인할 수 있었다. 면역조직화학적 염색의 결과, 5A5는 드물게 신경세포의 표면에서만 관찰되었는데 5A5로 염색되는 신경세포는 아세틸콜린 수용체 복합체와 연관되어 있지 않았다. 5B8은 신경세포 표면에서만 관찰되고 아세틸콜린 수용체 복합체가 형성된 부위에서도 관찰되어 신경세포가 근육세포와 접촉하여 근육신경연접을 형성하는 경우에는 접촉부위에 존재하는 신경세포유착물질이 일반적인 형태에서 sialylated 형으로 변화됨을 보여주어 신경세포 돌기들의 다발형성과 유사함을 알 수 있었다. 이상의 결과는 신경근육연접의 형성에는 신경세포와 근육세포의 유착이 중요하며, 신경세포유착물질은 신경근육연접의 형성에 직접적으로 관여한다기보다는 두 세포의 접촉을 촉진시킴으로써 근육신경연접에서 두 세포가 분리되지 않도록 하는 역할을 수행한다고 추정된다. Neuromuscular junction formation is one of the hot research area for understanding synapse formation, and the contact and adhesion between muscle and neurons during this procedure is regarded as one of important steps for synaptogenesis. The changes of neuronal cell adhesion molecules during nerve-muscle contats has not been revealed yet. In this study, we isolated skeletal muscle cells and ventral spinal cord neurons from Sprague-Dawley rats and observed the contact areas with a transmission electron microscpe and studied the presence of NCAM at the contact sites by immunohistochemistry. The ventral spinal cord neuronal processes contact intimately with skeletal muscle cells, some of which were submerged into the muscle surface and had synaptic vesicles. NCAM was expressed on neuronal processes, only sialylated form were associated with acetylcholine receptor aggregates. These results confirmed the significance of adhesion in neuromuscular junction formation and NCAM may participate in this process by preventing the separation of 2 cells at the contact site.

배양한 임신 말기 흰쥐 태자 가로무늬근육세포의 세포뼈대

엄창섭(Chang-Sub Uhm),이동호(Dong Ho Lee),김 현(Hyun Kim),서영석(Young-Suk Suh) 대한해부학회 1999 Anatomy & Cell Biology Vol.32 No.1

근육모세포의 유합과정 중에 일어나는 세포뼈대의 변화를 살펴보기 위하여 임신 말기 흰쥐에서 분리한 근육모세포를 전 자현미경용 금그리드 위에서 배양하고, 세포막과 세포질을 제거하여 세포뼈대 만을 남긴 후 투과형 전자현미경으로 관찰하였다. 방추형 근육모세포의 세포막 직하와 세포질 내에는 세포의 길이 방향으로 배열되어 있는 미세섬유의 다발이 존재하였고, 세포질 전체에는 망상으로 세포뼈대가 분포하고 있었다. 근육모세포의 표면돌기나 양단의 커텐형돌기의 기저부에는 미세섬유가 좁은 망상의 띠를 형성하고 있었으며, 이곳으로부터 방사상으로 미세섬유가 뻗어나가고 있었다. 세포체 부위에서 기시하는 돌기는 길이 방향으로 배열된 미세섬유의 다발에서 기시하는 경우도 있었다. 근육모세포의 접촉 부위에서는 인접한 세포를 연결시켜주는 가는 미세섬유 및 미세섬유망이 존재하고 있었으며, 길이 방향으로 배열된 미세섬유 다발이 서로 연결되어 있었다. 이상의 결과는 근육모세포의 세포뼈대가 이들 세포의 유합 과정에 따라 변화되고 있음을 보여주는 것이다. Myoblasts fuse together to form multinucleated myotubes. However, only a few studies have been reported on the cytoskeletal changes during the fusion process. To understand the change of cytoskeleton during the fusion process, isolated myoblasts from embryonic day 21 Sprague-Dawley rats were cultured on formvar-, carbon-, and gelatincoated gold grids for electron microscopy. The cells were fixed and plasma membrane and cytoplasm were extracted with triton X-100 and observed directly with Hitachi H-600 transmission electron microscope without staining. Fusiform myoblasts have complex cytoskeletal networks at the center of the cells, which were too dense to be resolved, however the margins of myoblasts and myotubes have bundles of cytoskeletons running in the longitudinal direction with reticulated cytoskeletal networks in between. Lamellipodial ruffles at both ends of myoblasts were characterized by a cytoskeletal lattice at the base and a few radiating strands into the filopodia-like processes. Radiating cytoskeletons originated either from the longitudinally oriented cytoskeletal bundles or reticular lattice continuous to them. The fusion areas were characterized by thin filaments connecting adjacent cells and the connection of longitudinal filament bundles from the fusing cells. These results suggest the modification of cytoskeletons during myoblast fusion.

한국인 신생남아에서 전내장전위증을 동반한 VATER증후군 1례

엄창섭(UHM Chang-Sub),신영철(SHIN Young-CHul) 대한체질인류학회 1988 해부·생물인류학 (Anat Biol Anthropol) Vol.1 No.1

Gemifloxacin이 미성숙 랫드의 아킬레스건에 미치는 영향

배진규,김영수,김세은,심경미,강성수,조익현,이수한,박창현,엄창섭,정문진,한송이,임성철,배춘식,Bae, Jin-Gye,Kim, Young-Soo,Kim, Se-Eun,Shim, Kyung-Mi,Kang, Seong-Soo,Cho, Ik-Hyun,Lee, Soo-Han,Park, Chang-Hyun,Uhm, Chang-Sub,Jeong, Moon-Jin,Han 한국현미경학회 2006 Applied microscopy Vol.36 No.2

Gemifloxacin은 대부분의 G(-) 세균들과 G(+) 세균들에 대해 강한 활성을 나타내며 미성숙동물에서 비교적 낮은 연골독성 가능성을 가진 합성 fluoroquinolone계 항생제이다. 본 연구는 미성숙 SD계 랫드의 아킬레스건에 gemifloxacin이 미치는 영향을 평가하기 위해 gemifloxacin을 생후 4주 후부터 0(vehicle), 600mg/kg body weight 용량으로 5일 동안 매일 한번씩 경구투여를 하여 ofloxacin과 비교하였다. 아킬레스건의 형태학적 변화를 투과전자현미경을 이용하여 관찰한 결과 부형제만 처치한 대조군과 비교하였을 때, gemifloxacin과 ofloxacin을 처치한 랫드로부터 채취한 샘플에서 초미세구조의 변화가 관찰되었다. quinolone을 처치한 군의 tenocyte에서는 다양한 퇴행성 변화가 관찰되었고, 세포외기질에서 분리된 세포들이 존재하였는데 이는 ofloxacin을 처치한 그룹에서 특히 높았고 gemifloxacin을 처치한 그룹에서는 매우 낮았다. 따라서, gemifloxacin이 아킬레스건과 같은 결합조직 구조물들의 퇴행성 변화를 보다 적게 야기한다는 것을 보여준다. Gemifloxacin is a synthetic fluoroquinolone antimicrobial agent that exhibits potent activity against most Gram-negative and Gram-positive organisms, and has a comparatively low chondrotoxic potential in immature animals. This study examined the effects of gemifloxacin on the Achilles tendons in immature Sprague-Dawley rats treated by oral intubation once daily for 5 consecutive days from postnatal week 4 onward at doses of 0 (vehicle), and 600mg/kg body weight Ofloxacin was used for comparison. The Achilles tendon sperimens were examined by electron microscopy. In comparison with the vehicle-treated controls, there were ultrastructural changes in all samples from the gemifloxacin- and ofloxacin-treated rats. Degenerative changes were observed in the tenocytes, and the cells that detached from the extracellular matrix were recognizable. The degree of degenerative changes and the number of degenerated cells in the Achilles tendon were significantly higher in the treated group than in the control group. Moreover, among the quinolone treated groups, these findings were more significant in the ofloxacin treated group, and less significant in the gemifloxacin treated group. It is unclear what these findings mean with respect to the possible risk ill juvenile patients treated with gemifloxacin or other quinolones. However, these results show that gemifloxacin causes fewer changes in the connective tissue structures.

흰쥐 부신수질 아민성세포의 분비과정에 관한 전자현미경적 관찰

류임주,엄창섭,서영석,Rhyu, Im-Joo,Uhm, Chang-Sub,Suh, Young-Suk 한국현미경학회 1992 Applied microscopy Vol.22 No.1

To clarify the exocytotic features in adrenal medullary aminergic cells, the authors observed rat adrenal medulla prepared by the TAGO method with transmission electron microscope. Rat adrenal medulla contains two types of aminergic cells, adrenergic and noradrenergic, as described. They were present as a group. In a single group both adrenergic and noradrenergic cells were present, but the same kind of cells showed the tendency forming small groups. Adrenergic cells were characterized with the granules having relatively electroluscent cores. These granules were relatively uniform in size, and the cores filled the granules with only thin halos. Noradrenergic cells were characterized with the granules of various size and forms. Most of the cores of these granules were generally more electron-dense than those of the adrenergic cells and only partly filled the granules without forming the halos. But, some granules were very similar in the shape and electron density as those of the adrenergic cells. Even empty-looking granules were present. Exocytotic figures with the classical omega figures were observed in both types of aminergic cells, but they were more frequent in adrenergic cells. These figures were mainly present along the plasma membranes toward the capillary. The excreted materials could be identified in the cleft of the omega figures. Apocrine-like secretory patterns but without cytoplasmic rims were identified in noradrenergic cells. Some vesicles, possibly formed from the cytoplsmic tubular systems were released. Some irregular lamellar structures of varying sizes were also observed. They looked like membranous structures sneaking through the plasma membranes. We could not, however, found any evidences of their involvement in exocytotic processes. These were present toward the capillaries and found only in the adrenergic cells. The authors conclude that the secretory processes in adrenal chromaffin cells may include not only the classical exocytotic processes but also the unusual direct secretions of granules or parts of cellular organelles. The membranous lamellar structures may indicate the remnants of excreted granules or functionally inactive excess membranes of the organelles removed from the cytoplasm.

한국산 박쥐 간장의 미세구조적 고찰

최병진,엄창섭,장병준,박창현,Choi, Byung-Jin,Uhm, Chang-Sub,Chang, Byung-Jun,Park, Chang-Hyun 한국현미경학회 2001 Applied microscopy Vol.31 No.4

한국산 관박쥐과의 관박쥐 (Rhinolophus ferrmmequinum korai)와 애기박쥐과 Myotis속에 속하는 큰발웃수염 박쥐 (Myotis macrodactylus), 물웃수염박쥐 (Myotis daubentonii ussrinesis) 및 관코박쥐 (Murina leucogaster intermedia)의 활동기 간장 조직을 투과전자현미경으로 관찰하여 이들의 미세구조적 특징을 규명하고 이들 간에 존재하는 차이를 비교 고찰하고자 하였다. 관박쥐는 세포질 내에 $10\sim20$개의 크고 둥근 사립체를 가지고 있었으며, 특히 과산화소체가 잘 발달되어 있었다. 큰발웃수염박쥐는 간세포간 사이의 연접복합체, 특히 부착반점이 잘 발달되어 있었다. 디세강과 동굴모세혈관 속은 섬세한 물질로 채워져 있었다. 물웃수염박쥐는 세포질 내에 당원과립을 많이 가지고 있었으며, 디세강과 동양혈관 속은 섬세한 물질로 채워져 있었다. 관코박쥐는 동굴모세혈관에 기저막이 잘 발달되어 있었으며, Kupffer 세포와 Ito 세포는 관찰되지 않았다. 이상의 결과는 같은 시기의 한국산 박쥐의 간장에서도 종에 따라서 미세구조적 차이가 나타남을 보여주는 것으로 이들은 이들의 생활환경과 식이와 관련이 있을 것으로 추정된다. To understand the fine structure of Korean bat livers , the authors investigated the liver of four Korean bats; Rhinolephus ferrumequinum korai, Myotis macrodactylus, Myotis dauhentonii ussrinesis, and Murina leucogaster intermedia by transmission electron microscopy. The hepatocytes of Rhinolophus ferrumequinum korai had large-sized mitochondria and many peroxisomes. In the Myotis macrodactylus, juctional completes, especially desmosomes, were well developed. The Myotis daubentonii ussrinesis had many glycogen particles in the cytoplasm. Also, the space of Disse and sinusoidal spare was filled with amorphous materials. In the Murina leucogaster intemedia, basement membrane was prominent in the sinusoid, and no Kupffer and Ito cells were observed These results suggest the characteristic differences in the liver ultrastructure among Korean bats.

허혈 및 재관류가 흰쥐 및 고양이 심장에 미치는 영향에 관한 형태계측학적 연구

박영식,엄창섭,서영석,Park, Young-Sik,Uhm, Chang-Sub,Suh, Young-Suk 한국현미경학회 1992 Applied microscopy Vol.22 No.1

To understand the structural changes of the myocardial myocytes and endothelial cells in ischemic and reperfused heart, and to elucidate their roles in those conditions, the authors observed cat and rat myocardium ultrastructurally and evaluated them with morphometric techniques. In cat, mild ischemia and moderate degree reperfusion injury was induced by ligation of the anterior interventricular branch of left coronary artery and reperfusion. In rat, severe ischemia and irreversible reperfusion iniury was made using in vitro Langendorff techniques. In normal cat myocytes, the volume densities of cytoplasm, myofibrils, mitochondria, sarcoplasmic reticulum and T tubules were $0.11{\pm}0.013,\;0.51{\pm}0.096,\;0.25{\pm}0.082,\;0.09{\pm}0.008,\;0.02{\pm}0.010$ (Mean${\pm}$S.D.) respectively, and the myofibril/mitochondria ratio was $2.33{\pm}1.379$. The numerical density and average volume of mitochondria were $0.76{\pm}0.210/{\mu}m^3$ and $0.33{\pm}0.057{\mu}m^3$ respectively. In normal cat endothelial cells, the volume densities of cytoplasm, cytoplasmic vesicles, tubular systems (including endoplasmic reticulum and Golgi apparatus) and mitochondria were $0.43{\pm}0.023,\;0.28{\pm}0.007,\;0.22{\pm}0.021,\;0.03{\pm}0.014$ respectively. The mean thickness of endothelial cells was $230{\pm}45.2{\mu}m$. The numerical density and average volume of cytoplasmic vesicles were $508{\pm}55.0/{\mu}m^3,\;578{\pm}104.8nm^3$ respectively. In cat myocytes which received mild ischemic injury, the volume densities of organelles were not changed significantly in ischemic and reperfusion states. In reperfusion group myocytes, the numerical density of mitochondria was decreased significantly and the average volume was increased significantly. In endothelial cells, the volume density of tubular system in ischemic group and the average volume of cytoplasmic vesicles in reperfusion group were increased significantly. In rat myocytes which received severe ischemic injury, the volume density and average volume of mitochondria were increased significantly, and the volume density of sarcoplasmic reticulum and numerical density of mitochondria were decreased significantly in both ischemic and reperfusion groups. In ischemic and reperfused endothelial cells, the volume density and numerical density of cytoplasmic vesicles, the volume density of cytoplasm were decreased significantly. The volume densities of tubular system were increased significantly in both ischemic and reperfused groups. The volume density of mitochondria in ischemic group and the average volume of cytoplasmic vesicles in reperfusion group showed significant increase. The authors, based on the above observations, conclude that the mitochondria of myocytes and the cytoplasmic vesicles of endothelia are the first group of targets in ischemic and reperfusion injury and in this respect, the degree of ischemic insult is not significant. The role of myocyte mitochondria in reperfusion injury may be insignificant, but endothelial cells may contribute actively to reperfusion injury.

흰쥐에서 분리 배양한 간세포의 담세관 형성에 있어서 액틴미세섬유의 역할에 관한 전자현미경적 연구

박창현,장병준,엄창섭,Park, Chang-Hyun,Chang, Byung-Joon,Uhm, Chang-Sub 한국현미경학회 1999 Applied microscopy Vol.29 No.4

간세포의 기능적 연구를 위한 배양계를 확립하기 위하여 Sprague-Dawley계 흰쥐의 간장에서 collagenase와 hyaluronidase의 혼합액을 이용하여 간세포를 분리하고 배양하여, 배양중인 간세포의 구조적인 변화와 담세관의 형성 과정을 확인하고, cytochalasin D를 배양계에 첨가한 경우 발생되는 간세포 및 담세관의 구조적인 변화를 살펴보아 다음과 같은 결과를 얻었다. 분리 배양한 흰쥐의 간세포는 원형이었고, 표면에 미세융모를 가지고 있었으며. 배양중 서로 부착되어 세포띠를 형성하였다. Cytochalasin D처리후 간세포의 표면은 미세융모가 소실되어 편평하게 변화되었으며, 소포성 돌출물이 자주 관찰되었다. 담세관은 부착된 간세포의 사이에서 형성되었으며, 간세포 표면의 작은 융기에서 기시하는 다양한 길이 및 형태의 미세융모로 채워져 있었고, 양단에는 치밀결합 및 부착만 등으로 구성된 연접복합체가 존재하였다. Cytochalasin D 처리후 당세관의 내강은 팽창되었으며 미세융모는 소실되어 거의 존재하지 많았고, 양단에 존재하는 연접복합체는 파괴되어 간격이 벌어진 곳이 많았다. 담세관내에 존재하는 미세융모 속에 존재하는 액틴미세섬유심은 완전하게 형성되어 있는 경우, 불완전하게 적은 양만 존재하는 경우, 그리고 전혀 존재하지 않는 경우가 있었다. 담세관주변세포질에 존재하는 액틴미세섬유얼기의 형성은 불완전하여 부위에 따라 없는 곳도 있었다. Cytochalasin D처리후 담세관주변세포질의 액면미세섬유얼기는 존재하지 많았다. 이상의 결과로 흰쥐의 간장에서 분리한 간세포는 배양중 성장하면서 정상적인 담세관을 형성함을 알 수 있었으며, 담세관의 형성은 접착부위의 연접복합체의 형성 및 미세융모의 형성,담세관 내 액틴미세섬유심 및 담세관주변세포질내 액틴미세섬유얼기의 형성 등을 특정으로 하는 것으로 판단된다. Bile canaliculi are the structure delivering bile secreted by hepatocytes into the bile passage. Bile secretion is mainly controlled by the cytoskeletal elements, mainly of actin in the microvilli, pericanalicular web. Most studies on the bile secretion have been done in viva situation, however, to control the various parameters in vitro culture system seem to be more useful. To set up an in vitro experimental system, the investigator isolated hepatocytes with an enzymatic method using a mixture of collagenase and hyaluronidase from normal Sprague-Dawley rat liver and cultured. Isolated hepatocytes were round and formed cords in culture. Microvilli covered the whole surface of hepatocytes. Bile canaliculi were formed between hepatocytes and were characterized by the presence of microvilli of various lengths and shapes mainly arising from small surface mounds. Actin filament core in the microvilli and pericanalicular actin web were incomplete. After cytochalasin D treatment, cultured hepatocytes were round but the surface were irregular with surfacen blebs, folds and grooves. Microvilli on the surface were scarce. Bile canaliculi were markedly dilated often with the detached junctional complexes. Bile canaliculi lacks microvilli almost completely and extended into the pericanalirular cytoplasm showing complex vacuolar and tubular structures by transmission electron mciroscopy. Pericanalicular actin web, intermediate filaments were hardly identified. Subsurface actin filaments were scattered scarcely under the cell membranes. These results suggest that hepatocytes isolated from rats can survive and form bile canaliculi in culture and the actin filaments are involved in the formation and/or maintenance of the bile canaliculi.