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Oral Administration of Lactilactobacillus curvatus LB-P9 Promotes Hair Regeneration in Mice

  • Mikyung Song;Jaeseok Shim;Kyoungsub Song
    • Food Science of Animal Resources
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    • v.44 no.1
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    • pp.204-215
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    • 2024
  • This study was designed to examine the effect of Lactilactobacillus curvatus LB-P9 on hair regeneration. The treatment of LB-P9 conditioned medium increased the proliferation of both hair follicle dermal papilla cells and hair germinal matrix cells (hGMCs). Moreover, the expression levels of hair growth factors such as vascular endothelial growth factor (VEGF) and fibroblast growth factor 7 were significantly elevated in hGMCs co-cultured with LB-P9. After time-synchronized depilation, mice were orally administered with either 4×107 colony forming unit (CFU) of LB-P9 (low dose) or 4×108 CFU of LB-P9 (high dose), once daily for 4 weeks. Compared with the vehicle (phosphate-buffered saline)-administrated group, the LB-P9-treated groups exhibited accelerated hair regrowth rate and enhanced hair thickness in a dose-dependent manner. Supporting this observation, both hair follicle numbers and the dermal thickness in skin tissues of the LB-P9-treated groups were increased, compared to those of the vehicle-treated group. These results might be explained by the increased level of β-catenin and number of hair follicle stem cells (CD34+ CD49f+ cells) in the skin tissues of mice administered with LB-P9, compared to the vehicle-treated mice. Also, increased serum levels of hair growth factors such as VEGF and insulin-like growth factor-1, and superoxide dismutase were found in the LB-P9-treated groups, compared to those of the vehicle-treated group. Taken together, these results might demonstrate that the oral administration of LB-P9 promotes hair regeneration by the enhancement of dermal papilla proliferation through the stimulation of hair growth factor production.

Early Changes after Death of Plaice, Paralichthys olivaceus Muscle -5. Effect of Storage Temperature on Morphological Changes of Myofibrils and Histological Changes of Muscle- (넙치(Paralichthys olivaceus)육의 사후 조기 변화 -5. 저장 온도가 근원섬유의 형태학적 및 육의 조직학적인 변화에 미치는 영향-)

  • CHO Young-Je;LEE Keun-Woo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.2
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    • pp.114-120
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    • 1994
  • To clarify the effect of storage temperature on the morphological and histological changes of plaice, Paralichthys olivaceus muscle at early stages after killing, the changes in breaking strength of muscle, morphological observation of myofibrils and histological observation of extracellular spaces during storage at $0^{\circ}C\;and\;10^{\circ}C$ were studied. The maximum breaking strength of samples stored at $0^{\circ}C$ was reached within 10hrs and then it dropped significantly (p<0.05) from 10hrs to 25hrs of storage. However, breaking strength was not increased in fresh muscle stored at $10^{\circ}C$ and gradually decreased after 10hrs storage. In myofibrils prepared from dorsal muscle immediately after death, A-band, I-band, H-band and Z-line in sarcomere were clearly distinguishable from each other. Due to muscle contraction, it was not easy to distinguish H-band from I-band observed in sarcomere stored at $0^{\circ}C$ after 10hrs storage. But, in the case of samples stored at $10^{\circ}C$, H-band could be observed dimly until 15hrs of storage. The changes in morphological myofibrils were closely related to increase of breaking strength. No extracellular space was observed among muscle cells immediately after killing. Stored samples at $0^{\circ}C$ showed extracellular spaces after 15hrs storage. On the other hand, samples stored at $10^{\circ}C$ didn't show any extracellular spaces until 15hrs storage and showed extracellular spaces after 24hrs storage. It was thought that the post-mortem tenderization of plaice muscle was closely related to the gradually disintegration of the extracellular matrix structure after killing.

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Estimate of Regional and Broad-based Sources for PM2.5 Collected in an Industrial Area of Japan

  • Nakatsubo, Ryouhei;Tsunetomo, Daisuke;Horie, Yosuke;Hiraki, Takatoshi;Saitoh, Katsumi;Yoda, Yoshiko;Shima, Masayuki
    • Asian Journal of Atmospheric Environment
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    • v.8 no.3
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    • pp.126-139
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    • 2014
  • In order to estimate the influence of sources on $PM_{2.5}$ in the industrial area of Japan, we carried out a source analysis using chemical component data of $PM_{2.5}$. $PM_{2.5}$ samples were collected intermittently at an industrial area in Japan from July 2010 to November 2012. Water soluble ions ($Cl^-$, $NO_3{^-}$, $SO{_4}^{2-}$, $Na^+$,$NH_4{^+}$, $K^+$, $Mg^{2+}$, $Ca^{2+}$), elements (Al, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, Sb, Pb), and carbonaceous species (OC, EC) of the $PM_{2.5}$ (a total of 198 samples) were analyzed. Positive Matrix Factorization (PMF) model was applied to the data of those chemical components to identify the source of $PM_{2.5}$. At this observation site, nine factors were extracted. The major contributors of $PM_{2.5}$ were secondary sulfate 1, in which loading factors of $SO{_4}^{2-}$ and $NH_4{^+}$ were large (percentage source contribution: 20.9%), traffic, in which loading factors of OC (organic carbon) and EC (elemental carbon) were large (20.8%), secondary sulfate 2, in which loading factors of K and $SO{_4}^{2-}$ were large (8.0%), steel mills (7.8%), secondary chloride and nitrate (7.0%), soil (5.0%), heavy oil combustion (3.8%), sea salt (3.8%), and coal combustion (2.3%). The conditional probability function (CPF) and the potential source contribution function (PSCF) were carried out to examine the influence of a regional source and a broad-based source, respectively. CPF results supported local source influences such as steel mills, sea salt, traffic, coal combustion, and heavy oil combustion. PSCF results suggested that ships in the East China Sea, an industrial area of the east coastal region of China, and an active volcano in the Kyushu region of Japan were potential regional sources of secondary sulfate 1. Secondary sulfate 2 was affected by the burning of biomass fields and by coal combustion in Chinese urban areas such as Beijing, Hebei, and western Inner Mongolia. Source characterization using continuous data from one site showed a potential source representing fossil fuel combustion is affected both by regional and broad-based sources.

A Study on the Ultrastructural Changes of Cardiac Muscle in Dichlorvos Treated Albino Rat (Dichlorvos가 흰쥐 심근의 미세구조에 미치는 영향)

  • Baik, Tai-Kyoung;Lee, Wha-Mo;Chung, Ho-Sam
    • Applied Microscopy
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    • v.24 no.3
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    • pp.23-33
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    • 1994
  • It is well known that dichlorvos (DDVP), an organophosphate insecticide in common use, is so easily and rapidly hydrolyzed and excreted that it has usually little toxic effect on human body. In these days, however, it is widely used as an industrial and domestic insecticide and as an anthelmintic agent for animals, so that the accident of chemical poisoning occurs frequently. DDVP acts as a powerful inhibitor of carboxylic esterase, which can cause accumulation of acetylcholine at the synapses so paralysis of muscle and the transmission failure in cholinergic synapses dueing to desensitization of acetylcholin receptor may occure. Moreover accumulation of the acetylcholine brings about the elevation of the cyclic-AMP, which alters the cellular metabolisms of nucleic acid, carbohydrate, protein and lipid. Present study has undertaken to investigate the cardiotoxic effect of DDVP by electron microscopic study. A total of 30 Sprague-Dawley strain rats, weighing about 250gm were used as experimental animals. 2mg/kg/day of DDVP is intraperitonealy injected 3 times with intervals of every other day. On 1 day, 3 days, 5 days, 7 days and 14 days after drug administration, the animals were sacrified by cervical dislocation. Left ventricular cardiac muscles were resected and sliced into $1mm^3$. The specimens were embedded with Epon 812 and prepared by routine methods for electron microscopical observation. All preparations were stained with lead citrate and uranyl acetate and then observed with Hitachi-600 transmission electron microscope. The results were as follows: 1. In the cardiac muscle of DDVP treated rats, mitochondria with disorganized double membrane and mitochondrial crista, and vacuole formation in mitochondrial matrix were observed. But structures of mitochondria were recovered to normal in 14 days group. 2. In the cardiac muscle of DDVP treated rats, cisternae of sarcoplasmic reticulum were dilated and sacculated. But these changes were recovered to normal in 14 days group. 3. In the cardiac muscle of DDVP treated rats, glycogen particles around damaged myofibrils were decreased. But amount of glycogen particles were restored in 14 days group. 4. In the cardiac muscle of DDVP treated rats, disruption and discontinuation of myofilaments and disorganization of Z-disc were observed. But the structures of myofibrils were recovered to normal in 14 days group. It is consequently suggested that DDVP would induce the reversible degenerative changes on the ultrastructures in cardiac muscle of rat.

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Beyond Clot Dissolution; Role of Tissue Plasminogen Activator in Central Nervous System

  • Kim, Ji-Woon;Lee, Soon-Young;Joo, So-Hyun;Song, Mi-Ryoung;Shin, Chan-Young
    • Biomolecules & Therapeutics
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    • v.15 no.1
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    • pp.16-26
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    • 2007
  • Tissue plasminogen activator (tPA) is a serine protease catalyzing the proteolytic conversion of plasminogen into plasmin, which is involved in thrombolysis. During last two decades, the role of tPA in brain physiology and pathology has been extensively investigated. tPA is expressed in brain regions such as cortex, hippocampus, amygdala and cerebellum, and major neural cell types such as neuron, astrocyte, microglia and endothelial cells express tPA in basal status. After strong neural stimulation such as seizure, tPA behaves as an immediate early gene increasing the expression level within an hour. Neural activity and/or postsynaptic stimulation increased the release of tPA from axonal terminal and presumably from dendritic compartment. Neuronal tPA regulates plastic changes in neuronal function and structure mediating key neurologic processes such as visual cortex plasticity, seizure spreading, cerebellar motor learning, long term potentiation and addictive or withdrawal behavior after morphine discontinuance. In addition to these physiological roles, tPA mediates excitotoxicity leading to the neurodegeneration in several pathological conditions including ischemic stroke. Increasing amount of evidence also suggest the role of tPA in neurodegenerative diseases such as Alzheimer's disease and multiple sclerosis even though beneficial effects was also reported in case of Alzheimer's disease based on the observation of tPA-induced degradation of $A{\beta}$ aggregates. Target proteins of tPA action include extracellular matrix protein laminin, proteoglycans and NMDA receptor. In addition, several receptors (or binding partners) for tPA has been reported such as low-density lipoprotein receptor-related protein (LRP) and annexin II, even though intracellular signaling mechanism underlying tPA action is not clear yet. Interestingly, the action of tPA comprises both proteolytic and non-proteolytic mechanism. In case of microglial activation, tPA showed non-proteolytic cytokine-like function. The search for exact target proteins and receptor molecules for tPA along with the identification of the mechanism regulating tPA expression and release in the nervous system will enable us to better understand several key neurological processes like teaming and memory as well as to obtain therapeutic tools against neurodegenerative diseases.

Studios on Intestinal Trematodes in Korea X. Scanning Electron Microscopic Observation on the Tegument of Fibricola seoulensis (한국의 간흡충에 관한 연구 X. Fibricola seoulensis 표피의 전자현미경적 관찰)

  • 서병설;이순향
    • Parasites, Hosts and Diseases
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    • v.22 no.1
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    • pp.21-29
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    • 1984
  • A scanning eletron microscopic study was performed to observe the tegumental surface of adult Fibricola seoulensis. The adult worms were collected from the small intestine of mice 5 days to 3 weeks after experimental infection with the metacercariae. The metacercariae were obtained from the viscera of the snakes, Matrix tigrina lateralis, by artificial digestion technique. The results were as follows: 1. The tegument of anterior body was covered with cobblestone-like cytoplasmic processes and that of posterior body showed finger-like processes. The posterior body had 4-5 large transverse wrinklings which formed many discontinued shallow rugae. 2. The entire surface of anterior body was regularly arranged with the spines of which tips diverged into 3 to 4 points. They were densely packed in anterior mid-median portion of dorsal surface where appeared a few spines indented upto 5 points. Farther laterally and posteriorly from this portion, the pointed spines were more sparse and became single tipped and extended to anterior one-third of posterior body, 3. The posterior surface of oral sucker was armed with 50-60 spines having 2-3 tips and ventral sucker also covered with such spines. On anteriormost dorsal surface arranged 60-70 spade-shaped spines. The tribocytic organ was armed with many stout recurved pile-like spines arranged radially. 4. There were 3 types of sensory papillae. The ciliated knob-like (Type I) papillae were almost bilaterally symmetrical in ventral and dorsal surfaces of anterior body, and abundant especially aroundbases of oral and ventral suckers, tribocytic organ, and in lateral margins of anterior body. About 24 non-ciliated round swellings (Type II) were observed around each lip of oral and ventral suckers. The plate-like elevated papilla without cilium (Type III) was found to distribute only in posterior body. These 3 types of papillae seem to be tangoreceptive and/or rheoreceptive in function when their morphology and distributions are considered.

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Postchilling Accumulation of Superoxide in Cells and Chilling Injury in Rice Plant (Superoxide의 세포내 축적과 벼냉해의 발현)

  • Kim, Jong-Pyung;Hyun, Il;Jung, Jin
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.364-370
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    • 1987
  • The $O_2^-$ level of the extract from young rice leaves, which was cold treated for 2 days and then placed at room temperature for a period of time significantly higher than that from tissues untreated. $O_2^-$ level in leaves was practically unchanged during cold treatment for 48 hours. But it started to increase to arrive at maximum in 8 hours, once the plants were placed under room temperature. The abnormal production of $O_2^-$ in mitochondria during postchilling process was interpreted as a biochemical consequence of accumulation of glycolysis product(s) in cytosol and/or NADH in mitochondrial matrix due to disruption of catabolic balance at low temperature. Mitochondria isolated from the chilling injured tissue was found to have lost considerably their respiratory activity. This fact may imply the involvement of intramitochondrial accumulation of $O_2^-$ in the inactivation of electron transport chain system. The observation that mitochondria in the presence of the $O_2^--producing$ enzymatic system (Xanthine/Xanthine oxidase) lost their respiratory activity supports this inference. It was also found in this work that Superoxide dismutase (SOD) is a substrate inducible enzyme, and that SOD is a possible protective agent in plant cell against chilling injury.

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Effects of Tiam 1 on Invasive Capacity of Gastric Cancer Cells in vitro and Underlying Mechanisms

  • Zhu, Jin-Ming;Yu, Pei-Wu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.201-208
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    • 2013
  • Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

"Over-inlay" block graft and differential morphometry: a novel block graft model to study bone regeneration and host-to-graft interfaces in rats

  • Ghiacci, Giulia;Graiani, Gallia;Ravanetti, Francesca;Lumetti, Simone;Manfredi, Edoardo;Galli, Carlo;Cacchioli, Antonio;Macaluso, Guido Maria;Sala, Roberto
    • Journal of Periodontal and Implant Science
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    • v.46 no.4
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    • pp.220-233
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    • 2016
  • Purpose: The aim of this study was to present new a model that allows the study of the bone healing process, with an emphasis on the biological behavior of different graft-to-host interfaces. A standardized "over-inlay" surgical technique combined with a differential histomorphometric analysis is presented in order to optimize the use of critical-size calvarial defects in pre-clinical testing. Methods: Critical-size defects were created into the parietal bone of 8 male Wistar rats. Deproteinized bovine bone (DBBM) blocks were inserted into the defects, so that part of the block was included within the calvarial thickness and part exceeded the calvarial height (an "over-inlay" graft). All animals were sacrificed at 1 or 3 months. Histomorphometric and immunohistochemical evaluation was carried out within distinct regions of interest (ROIs): the areas adjacent to the native bone (BA), the periosteal area (PA) and the central area (CA). Results: The animals healed without complications. Differential morphometry allowed the examination of the tissue composition within distinct regions: the BA presented consistent amounts of new bone formation (NB), which increased over time ($24.53%{\pm}1.26%$ at 1 month; $37.73%{\pm}0.39%$ at 3 months), thus suggesting that this area makes a substantial contribution toward NB. The PA was mainly composed of fibrous tissue ($71.16%{\pm}8.06%$ and $78.30%{\pm}2.67%$, respectively), while the CA showed high amounts of DBBM at both time points ($78.30%{\pm}2.67%$ and $74.68%{\pm}1.07%$, respectively), demonstrating a slow remodeling process. Blood vessels revealed a progressive migration from the interface with native bone toward the central area of the graft. Osterix-positive cells observed at 1 month within the PA suggested that the periosteum was a source of osteoprogenitor elements. Alkaline phosphatase data on matrix deposition confirmed this observation. Conclusions: The present model allowed for a standardized investigation of distinct graft-to-host interfaces both at vertically augmented and inlay-augmented sites, thus possibly limiting the number of animals required for pre-clinical investigations.

A STUDY ON THE RESPONSES OF OSTEOBLASTS TO VARIOUS SURFACE-TREATED TITANIUM

  • Lee Joung-Min;Kim Yung-Soo;Kim Chang-Whe;Jang Kyung-Soo;Lim Young-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.307-326
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    • 2004
  • Statement of problem. The long-term success of implants is the development of a stable direct connection between bone and implant surface, which must be structural and functional. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. Among them, altering the surface properties can modify cellular responses such as cell adhesion, cell motility and bone deposition. Purpose. This study was to evaluate the cellular behaviors on the surface-modified titanium by morphological observation, cellular proliferation and differentiation. Material and methods. Specimens were divided into five groups, depending on their surface treatment: electropolishing(EP) anoclizing(AN), machining(MA), blasting with hydroxyapatite particle(RBM) and electrical discharge machining(EDM). Physicochemical properties and microstructures of the specimens were examined and the responses of osteoblast-like cells were investigated. The microtopography of specimens was observed by scanning electron microscopy(SEM). Surface roughness was measured by a three-dimensional roughness measuring system. The microstructure was analyzed by X-ray diffractometer(XRD) and scanning auger electron microscopy(AES). To evaluate cellular responses to modified titanium surfaces, osteoblasts isolated from neonatal rat were cultured. The cellular morphology and total protein amounts of osteoblast-like cell were taken as the marker for cellular proliferation, while the expression of alkaline phosphatase was used as the early differentiation marker for osteoblast. In addition, the type I collagen production was determined to be a reliable indicator of bone matrix synthesis. Results. 1. Each prepared specimen showed specific microtopography at SEM examination. The RBM group had a rough and irregular pattern with reticulated appearance. The EDM-treated surface had evident cracks and was heterogeneous consisting of broad sheet or plate with smooth edges and clusters of small grains, deep pores or craters. 2. Surface roughness values were, from the lowest to the highest, electropolished group, anodized group, machined group, RBM group and EDM group. 3. All groups showed amorphous structures. Especially anodized group was found to have increased surface oxide thickness and EDM group had titaniumcarbide(TiC) structure. 4. Cells on electropolished, anodized and machined surfaces developed flattened cell shape and cells on RBM appeared spherical and EDM showed both. After 14 days, the cells cultured from all groups were formed to be confluent and exhibited multilayer proliferation, often overlapped or stratified. 5. Total protein amounts were formed to be quite similar among all the group at 48 hours. At 14 days, the electropolished group and the anodized group induced more total protein amount than the RBM group(P<.05). 6. There was no significant difference among five groups for alkaline phosphatase(ALP) activity at 48 hours. The AN group showed significantly higher ALP activity than any other groups at 14 days(P<.05). 7. All the groups showed similar collagen synthesis except the EDM group. The amount of collagen on the electropolished and anodized surfaces were higher than that on the EDM surface(P<.05).