• Title/Summary/Keyword: $CO_2$-cut

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Effects of SIS/PLGA Porous Scaffolds and Muscle-Derived Stem Cell on the Formation of Tissue Engineered Bone (SIS/PLGA 담체와 근육유래 줄기세포를 이용한 생체조직공학적 골재생)

  • Kim Soon Hee;Yun Sun Jung;Jang Ji Wook;Kim Moon Suk;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.14-21
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    • 2006
  • Tissue engineering techniques require the use of a porous biodegradable/bioresorbable scaffold, which server as a three-dimensional template for initial cell attachment and subsequent tissue formation in both in vitro and in vivo. Small intestinal submucosa (SIS) has been investigated as a source of collagenous tissue with the potential to be used as biomaterials because of its inherent strength and biocompatibility. SIS-loaded poly(L-lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/particle leaching. Characterizations of SIS/PLGA scaffold were carried out by SEM, mercury porosimeter, and so on. Muscle-derived stem cells can be differentiated in culture into osteoblasts, chondrocytes, and even myoblasts by the controlling the culture environment. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by alkaline phosphatase(ALP) activity. SIS/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of SIS on the osteoinduction compared with controlled PLGA scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were conducted hematoxylin and eosin (H&E), Trichrome, and von Kossa. We observed that bone formatioin of SIS/PLGA hybrid scaffold as natural/synthetic scaffold was better thean that of only PLGA scaffold. It canb be explained that SIS contains various kinds of bioactive molecules for osteoinduction.

Hypoxia Induced Expression of Vascular Endothelial Growth Factor in Rat Pulmonary Artery Smooth Muscle Cells (쥐의 폐동맥 평활근 세포에서 저산소에 의한 Vascular Endothelial Growth Factor의 발현)

  • Nho, Un Seok;Kim, Yeo Hyang;Hyun, Myung Chul;Lee, Sang Bum
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.167-172
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    • 2003
  • Purpose : Pulmonary vascular hypertension is a common problem in congenital heart disease, the most common cardiac condition in childhood. However, the mechanisms responsible for this pathologic change, treatment, and prevention are poorly understood. Therefore, we studied the gene expression of vascular endothelial growth factor(VEGF) by using a hypoxic model of the pulmonary artery smooth muscle cells. Methods : The main pulmonary artery and its proximal branches of a 6 wk old Fischer rat were excised. They were cut into multiple small pieces and suspended in DMEM medium supplemented with 20% fetal bovine serum and incubated in 5% $CO_2$-95% air atmosphere. The smooth muscle cells were confirmed by immunostaining with smooth muscle myosin and ${\alpha}$-smooth muscle actin antibodies. The VEGF gene expression in the hypoxic group was compared with the one in control the group as well as the one in the starved group by RT-PCR and Northern blot hybridization. Results : There was no statistically significant difference among the control, hypoxic and starved groups. Conclusion : There are few studies of pulmonary vascular hypertension at the molecular level in Korea. Therefore, we studied the expression of VEGF gene in hypoxic pulmonary vascular smooth muscle cells. Further studies will be needed to find the difference between newly born and adult rats, or human and rat pulmonary vascular smooth muscle cells in gene expression. We hope that the study will lead to a better understanding of pulmonary vascular hypertension.

The Characteristics for BNCT facility in Hanaro Reactor

  • Soheigh Suh;Lee, Dong-Han;Ji, Young-Hoon;Lee, Dong-Hoon;Yoo, Seong-Yul;Rhee, Chang-Hun;Rhee, Soo-Yong;Jun, Byung-Jin
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.161-163
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    • 2002
  • The BNCT(Boron Neutron Capture Therapy) facility has been developed in Hanaro(High-flux Advanced Neutron Application Reactor), a research reactor of Korea Atomic Energy Research Institute. A typical tangenial beam port is utilized with this BNCT facility. Thermal neutrons can be penetrated within the limits of the possible maximum instead of being filtered fast neutrons and gamma rays as much as possible using the silicon and bismuth single crystals. In addition to, the liquid nitrogen (LN$_2$) is used to cool down the silicon and bismuth single crystals for the increase of the penetrated thermal neutron flux. Neutron beams for BNCT are shielded using the water shutter. The water shutter was designed and manufactured not to interfere with any other subsystem of Hanaro when the BNCT facility is operated. Also, it is replaced with conventional beam port plug in order to cut off helium gas leakage in the beam port. A circular collimator, composed of $\^$6/Li$_2$CO$_3$ and polyethylene compounds, is installed at the irradiation position. The measured neutron flux with 24 MW reactor power using the Au-198 activation analysis method is 8.3${\times}$10$\^$8/ n/cm$^2$ s at the collimator, exit point of neutron beams. Flatness of neutron beams is proven to ${\pm}$ 6.8% at 97 mm collimator. According to the result of acceptance tests of the water shutter, the filling time of water is about 190 seconds and drainage time of it is about 270 seconds. The radiation leakages in the irradiation room are analyzed to near the background level for neutron and 12 mSv/hr in the maximum for gamma by using BF$_3$ proportional counter and GM counter respectively. Therefore, it is verified that the neutron beams from BNCT facility in Hanaro will be enough to utilize for the purpose of clinical and pre-clinical experiment.

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Fertilizing Ability of Bovine Spermatozoa Following Oviduct Epithelial Cell Co-culture In Vitro (난관상피세포와 공배양한 소 정자의 체외수정능)

  • 황우석;노상호;이병천
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.227-233
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    • 1998
  • The aim of these experiments was to investigate the effects of oviduct epithelial cells on bovine in vitro fertilization. Oviduct epithelial cell monolayers (OEC) on the 4-well dish were prepared according to general procedures. Monolayers were formed within 5days. The medium for OEC culture (TCM199 with 10% FBS) was replaced with IVF-TALP 2h before each experiment. Macromolecules/proteins from oviductal conditioned medium (OM) were recovered by ultrafiltration, which desalted and concentrated macromolecules greater than 5kDa, and this OM were added to W medium (experiment 1). The cleavage rate in OM+OEC group was significantly higher than in OM group (p〈0.01). In this experiment 2, oocytes were inseminated on OEC with sperm which had been pre-incubated with OEC for 0 or 4h before insemination. In this experiment, oocytes were exposed to sperm only 8 h for clarifying the effect. After insemination, oocytes were cultured in CRlaa. At 42 h post insemination, oocytes were denuded and examined for evidence of cleavage. The cleavage rates of oocytes which were inseminated with OEC treated sperm for 4 h were significantly higher than those of the other group (p〈0.01). In conclusion, sperm released from OEC have more fertilizing ability than those before attachment.

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Development of Effective Screening Method for Efficacy Test of Trunk Injection Agents Against Pine Wood Nematode, Bersaphelenchus xylophilus in Japanese Black Pine, Pinus thunbergii (곰솔(Pinus thunbergii)에서 소나무재선충(Bersaphelenchus xylophilus) 나무주사 약제의 효율적인 평가법 개발)

  • Shin, Won Sik;Jung, Young Hack;Lee, Sang Myeong;Lee, Chae Min;Lee, Chang Joon;Kim, Dong Soo;Mun, Il Sung;Lee, Dong Woon
    • The Korean Journal of Pesticide Science
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    • v.19 no.4
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    • pp.440-449
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    • 2015
  • Pine wood nematode (PWN), Bursaphelenchus xylophilus is one of the most serious pests of pine tree. Trunk injection of some nematicides in tree is well known as an effective control method. However there are some limitating factors which hindering the efficacy of trunk injection in field or potted tree. In this study we suggested easy and useful alternative screening methods of nematicides against PWN. Reproduction of PWN was influenced by tree twig moisture (high reproduction in high moisture trig) and paraffin coating was 78.6% reduced moisture loss in tested twig. There were no reproduction different in up and down site from infection site of twig at 1 month after inoculation of PWN and also distance (5 and 10 cm) from inoculation site of PWN did not influence the reproduction of PWN. Numbers of reproduced PWN were higher with decreasing diameter of twig. Numbers of reproduced PWN were similar to P. thunbergii and P. densiflora. However reproduction was increased depending on high inoculation density and longer propagation period. When inoculation of PWN on cut twig injected with emamectin benzoate 2.15% EC and morantel tartrate 8% SL in trunk of Pinus thunbergii in the field, PWN number were significantly reduced than untreated control. We suggest this screening method for PWN control agents.

Experimental Studies on the Effects of Pyeongpaesan (평폐산(平肺散)의 효능(效能)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Lee, Cheol-Hyeon;Shin, Jo-Young
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.385-408
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    • 1998
  • Pyeongpaesan (平肺散) has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Pyeongpaesan (平肺散) on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Pyeongpaesan (平肺散) on histamine and acetylcholine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig (500 g, male) and Sprague Dawley rats (200 g, male) were killed by $CO_2$ exposure and a segment (8-10 mm) of the thoracic trachea from each rat and guinea pig was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 0.5 g loading tension. The dose of histamine (His) and acetylcholine (Ach) which evoked 50% of maximal response ($ED_{50}$) was obtained from cumulative dose response curves for histamine and acetylcholine $(10^{-7}{\sim}10^{-4}M)$. Contractions evoked by His $(ED_{50})$ and Ach $(ED_{50})$ were inhibited significantly by Pyeongpaesan (平肺散). In guinea pig tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was 13.5% (p<0.05) after $10{\mu}l/ml$ Pyeongpaesan (平肺散), $64.6\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $92.8\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). In rat tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $60.9\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $91.2\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). Also, in guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $104.8\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散) and $142.3\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $63.7\(p<0.01)\;after\;30{\mu}l/ml$ Pyeongpaesan (平肺散), and $107.5\(p<0.01)\;after\;100{\mu}l/ml$ Pyeongpaesan (平肺散). Propranolol $(10^{-7}M)$ slightly but significantly attenuated the inhibitory effects of Pyeongpaesan (平肺散). Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 15.7% (p<0.05) in guinea pig induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 22.3% (p<0.05) in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Pyeongpaesan (平肺散) fell to 28.7% (p<0.01) in rat induced by histamine contraction. Indomethacin and methylene blue $(10^{-7}\;M)$ did not significantly alter the inhibitory effect of Pyeongpaesan (平肺散). Also, I could find the effects of Pyeongpaesan (平肺散) and Pyeongpaesanga (平肺散加) morphine on the tracheal smooth muscle in guinea pig and rat did not change significantly. These results indicate that Pyeongpaesan. (平肺散) can relax histamine and acetylcholine-induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects and the release of cyclooxygenase products.

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Study on the Effect of Jakyakgamcho-tang (작약감초탕의 효능에 관한 연구)

  • Lee Yun Suk;Kim Hyung Chang;Whang Eui Hyun;Jo Sung Kyun;Lim In Gyu;Han Jong Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.493-498
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    • 2003
  • Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamchotang have been used in Oriental Medicine for many centuries as a treatment for various disease. The purpose of the present study is to determine the effect of Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamchotang on narepinephrine(NE) induced contraction of isolated rabbit femoral artery. Rabbits (2.0kg, female) were killed by CO₂ exposure and a segment (8-10mm) of the aortic ring from each rabbit was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 1.5g loading tension. The dose of NE which evoked 50% of maximal response (ED/sub 50/) was obtained from cumulative dose response curves for NE (10/sup -7/~10/sup -4/M). Contractions evoked by NE (ED/sub 50/) were inhibited significantly by Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamcho-tang. The mean percent inhibition of NE induced contraction was 83.9% (p<0.01) after 150㎕/㎖ Paeoniae Radix, 101.1 %(p<0.01) after 150 ㎕/㎖, Glycyrrhizae Radix and 107.3%(p<0.01) after 150㎕/㎖ Jakyakgamcho-tang, Indomethacin slightly but significantly attenuated the inhibitory effects of Paeoniae Radix. Following treatment with indomethacin, the mean percent inhibition caused by 150㎕/㎖ Paeoniae Radix fell to 16.4% in femoral artery induced by NE contraction. Propranolol, ODQ, and L-NNA did not significantly alter the inhibitory effect of Paeoniae Radix. ODQ slightly but significantly attenuated the inhibitory effects of Glycyrrhizae Radix. Following treatment with ODQ, the mean percent inhibition caused by 150㎕/㎖ Glycyrrhizae Radix fell to 13.0% in femoral artery induced by NE contraction. Propranolol, indomethacin and L-NNA did not significantly alter the inhibitory effect of Glycyrrhizae Radix. L-NNA slightly but significantly attenuated the inhibitory effects of Jakyakgamchotang. Following treatment with L-NNA, the mean percent inhibition caused by 150㎕/㎖ Jakyakgamchotang fell to 13.8% in femoral artery induced by NE contraction. Propranolol, ODQ and indomethacin did not significantly alter the inhibitory effect of Jakyakgamcho-tang. These results indicate that Paeoniae Radix, Glycyrrhizae Radix and Jakyakgamcho-tang can relax NE induced contraction of the isolated rabbit femoral artery, and that this inhibition related to nitric oxide.

Desorption Kinetics and Structural Changes of Tetracycline Treated Barrier Membranes for Guided Tissue Regeneration (Tetracycline 처리된 조직유도재생술용 차폐막의 약제유리양상 및 구조적 변화)

  • Lee, Sung-Mi;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.27 no.1
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    • pp.1-17
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    • 1997
  • Tetracycline is known to be effective in eliminating periodontopathogens and have collagenolytic activity. This study was performed to observe the desorption kinetics and structural changes of tetracycline-treated barrier membranes for guided tissue regeneration. Four kinds of barrier membranes were tested : $Tefgen^{(R)}$(American Custom Medical, USA) and $Gore-Tex^{(R)}$(W.L. Gore & Associates Inc., USA) as nonresorbable membranes ; Resolut(polyglycolide & polylactide copolymer, W.L. Gore & Associates Inc., USA) and $Biomend^{(R)}$(collagen, Collatec Co., USA) as resorbable membranes. The membranes were cut into discs(diameter : 4mm) and were immersed in 5% tridodecylmethylammonium chloride(TIMAC) ethanol and air-dried. The membrane discs were absorbed with $100{\mu}g/ml tetracycline solution(pH8) for one minute and dried. For desorption kinetics, TC treated discs were immersed in phosphate buffered saline solution (PBS, pH 7.4). PBS was exchanged daily and TC concentration was measured by absorbance at 276nm on UV spectrophotometer. To measure remaining antibacterial activity, discs of 1 day to 4 weeks after desorption were placed on Mueller Hinton agar containing Bacillus cereus and incubated aerobically in $37^{\circ}C$ for twelve hours and the inhibition diameters were measured. To observe the structural change of membranes after TIMAC treatment or immersion in PBS, the membrane discs were examined under SEM. The results were as follows : 1. Total amounts of TC absorbed into membrane discs($0.7536mm^2$) were $2000{\mu}g$, $1800{\mu}g$, $2625{\mu}g$ and $2499{\mu}g$ for $Tefgen^{(R)}$, $Gore-Tex^{(R)}$, $Biomend^{(R)}$ and $Resolut^{(R)}$. 2. The concentration of TC released from barrier membrane discs was maintained over $4{\mu}g/ml$ until the fifth day in nonresorbable membranes and $Resolut^{(R)}$, but until the fourth day in $Biomend^{(R)}$, Until the ninth day in nonresorbable membranes and until the seventh day in resorbable membranes, the TC concentration was maintained over $1{mu}g/ml$. 3. The four membrane discs in the first day showed similar size of inhibition zone. One to four weeks later, the inhibition zone was much smaller in resorbable membrane discs than nonresorbable membrane discs. 4. Any structural change due to treatment of TIMAC was not observed on the nonresorbable membranes. $Resolut^{(R)}$ did not show any structural change except fibrillar loosening during immersion period, but Biomend showed destruction of membrane structure from the first week of immersion. This study indicates that tetracycline treated barrier membranes lead to the sustained release of tetracycline for over 7 days. This slow release pattern of tetracycline may contribute to the favorable clinical outcome of guided tissue regeneration.

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A STUDY ON THE CORROSION BEHAVIOUR OF AMALGAMS BY USING OF POTENTIOSTATIC POLARIZATION METHOD (정전압 분극곡선법에 의한 아말감의 부식 거동에 관한 연구)

  • Shin, Dong-Hoon;Um, Chung-Moon
    • Restorative Dentistry and Endodontics
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    • v.12 no.1
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    • pp.39-49
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    • 1986
  • The purpose of this study was to observe characteristic properties through the polarization curves and SEM images from 4 different types of amalgam obtained by using the potentiostats (Princeton EG & G) & SEM (Jeol/35), and to investigate the degree of corrosion on the oxidation peak of the each phase of amalgam. After each amalgam alloy and Hg was triturated as the direction of the manufacturer by means of the mechanical amalgamator (Shofu Co.), the triturated mass was inserted into the cylindrical metal mold which was 12mm in diameter and 10.0mm in height and was condensed by using routine manner. The specimen was removed from the mold and stored at room temperature for about 24 hours. The standard surface preparation was routinely carried out. Anodic polarization measurement was employed to confirm the corrosion behaviour of the amalgams in a 0.9% saline solution (P.H: 6.8-7.0) at $37^{\circ}C$. The initial rest potential (corrosion potential) was determined after 30 minutes of immersion of specimen in electrolyte, and the potential scan was begun at the point of 100mV cathodic from the corrosion potential. The scan rate was 0.17mV/sec. in the study to observe the degree of corrosion of each phase. SEI and EPMA images on the determined oxidation peaks of each amalgam were observed. The results were as follows: 1. In the four anodic polarization curves, low copper amalgams have three oxidation peaks and high copper amalgams have two oxidation peaks, -270mV, +26mV and +179mV(SEC) in the low copper lathe cut, and -300mV, +39mV and +163mV(SEC) in the low copper spherical. -4mV and +154mV(SEC) in the Dispersalloy, and +17mV and +180mV(SEC) in the Tytin as high copper amalgams. 2. ${\gamma}_2$ phase in the low copper amalgam and ${\eta}$ phase in the high copper amalgam were the most corrodible phases and Ag-Cu eutectic in high copper amalgam was the most slowly corroded phase. 3. Low copper amalgam was more susceptible in corrosion than high copper amalgam.

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Effect of Root Surface Treatment Using EDTA on the Initial Attachment of Human Gingival Fibroblasts (EDTA를 이용한 치근면 처리가 치은섬유모세포의 초기 부착에 미치는 영향)

  • Kim, Seong-Bong;Lim, Ki-Jung;Kim, Sang-Mok;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.30 no.1
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    • pp.145-157
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    • 2000
  • Cytotoxic substances in dental calculus and root cementum of periodontally diseased teeth inhibit new attachment and regeneration. The purpose of scaling and root planing is to remove pathologic structures harboring these cytotoxic substances in order to create a biologically acceptable root surface. However, these procedures inevitably leave a non-biocompatible smear layer. Conventionally, the smear layer has been removed with low pH etching agents such as citric acid, phosphoric acid and tetracycline hydrochloride(TC). Lately, a supersaturated neutral pH etching solution of ethylene diamine tetraacetic acid(EDTA) has been found to be as effective as low pH etchants with respect to smear removal and to be superior in exposing root surfaceassociated collagen. The aim of the present study was to determine the effect of root surface treatment using EDTA on the initial attachment of human gingival fibroblasts. 27 human teeth, extracted due to severe periodontitis, were cut into dentin slices after root planing. The specimens were divided into TC group(treated with $50㎎/m{\ell}$ tetracycline-HCl, pH 1.52), EDTA group(treated with 17% EDTA, pH 7.4), and non-treated control group. After sterilization, 5th subcultured human gingival fibroblasts were seeded in each culture well containing a prepared root slice and incubated for 15 min., 60 min., and 4 hours in 5% $CO_2$ incubator at $37^{\circ}C$. At each incubation time, the number of attached fibroblasts were counted on the microphotographs taken at a magnification of x100. The difference of the number of attached cells between groups was statistically analyzed by the ANOVA followed by Duncan test in SPSS/PC+programs. The results were as follows : 1. After incubation for 15 min, the attached cells were significantly more in EDTA group and TC group than non-treated control group(p<0.05), but there was no significance in the difference between EDTA group and TC group(p>0.1). 2. After incubation for 60 min and 4 hours, there was no significant difference in the number of attached cells between all groups(p>0.1). 3. In both EDTA group and TC group, there was no significant difference in the number of attached cells between different incubation(p>0.1). But in control group, the number of attached cells was significantly increased after incubation for 60 min, compared with incubation for 15 min(p<0.05). The above results suggest that root surface treatment using EDTA could enhance the initial attachment of gingival fibroblasts to root surface as effective as tetracycline-HCl.

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