• Title/Summary/Keyword: Pulp cell

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COMPARATIVE STUDY ON THE DETECTION OF PORPHYROMONAS ENDODONTALIS BY ANAEROBIC CULTURE, IIF AND DNA PROBE METHOD IN INFECTED ROOT CANALS (감염 근관에서 혐기성 배양법과 간접 면역 형광법 및 DNA 프로브법에 의한 Porphyromonas endodontalis의 검출에 관한 비교 연구)

  • Kim, Min-Kyum;Yoon, Soo-Han;Chung, Chong-Pyoung
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.1-18
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    • 1996
  • There are many advantages when using IIF and DNA probe methods over anaerobic culture method in that they are time-and effort-saving, more precise and more sensitive. Furthermore, in IIF and DNA probe methods, the detection is possible only with small amount of bacteria, the quantitative analysis is possible, and the cell viability is not necessary. The purpose of this study is to observe the incidence of P.endodontalis by carrying out anaerobic culture, IIF and colony lift using DNA probe method respectively, and to compare these 3 methods in terms of effectiveness and sensitivity in order to identify the most effective detection method. 30 teeth with at least one clinical symptoms, with single canal, and with pulp necrosis were sampled. For sampling bacteria, access cavity was prepared after disinfecting tooth and its surroundings. Then the paper point was inserted up to the periapical area, leave there for a while, and finally it was placed into PRAS Ringer's sol. and PBS sol. In anaerobic culture method, P.endodontalis was identified by biochemical tests after subculturing black and brown colonies which were produced after 7 days of incubation on BAP and Brucella BAP in anaerobic chamber. To identify P.endodontalis in IIF method, species-specific polyclonal rabbit-antisera of P.endodontalis(ATCC 35406) was reacted with sampled PBS sol. dispensed onto glass slide, and then P.endodontalis was examined by phase contrast microscopy after incubating with Goat anti-rabbit lgG conjugated to Fluorescein isothiocyanate. For colony lift using DNA probe method, membranes were laid over colonies on the surface of BAP and were hybridized with cloned DNA probe of P.endodontalis. The existence of P.endodontalis was then identified by the methods of chemiluminescent detection and color metric detection. Black colony was found in 11 teeth out of 30 teeth and P.endodontalis was detected in 6 teeth (20 %) by anaerobic culture method, 16 teeth (53 %) by IIF method, and 7 teeth (23 %) by DNA probe method. IIF method is significantly better in detecting P.endodontalis than DNA probe method and anaerobic culture method. There was no significant differences between DNA probe method and anaerobic culture method. There was significant correlation between the formation of black colony and the existence of P.endodontalis. The probability of detecting P.endodontalis when black colony being present is 2.89 times higher than when not being present. There was significant relationship between the foul odor of clinical symptoms and P.endodontalis. The sensitivity of existing P.endodontalis when foul odor being present was 93.75 %, while the specificity of not existing P.endodontalis when foul odor not being present was 28.57 %. These results suggested that the probes of P.endodontalis will be used to decide the method and prognosis in endodontic treatments.

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AN EXPERIMENTAL STUDY OF THE DEGREE OF CONVERSION AND CYTOTOXICITY OF DUAL CURE RESIN CEMENTS (수종 이원중합 레진 씨멘트의 중합률 및 세포 독성에 관한 실험적 연구)

  • Roh, Byoung-Duck;Park, Seong-Ho;Lee, Chung-Suck
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.33-54
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    • 1995
  • The degree of conversion of composite resin was known to have influence on the mechanical properties of composite materials such as hardness, strength, wear resisitance, dimensional and color stability. Also unreacted monomer was reported to be harmful to the pulp. So the degree of conversion was a very important factor in the success of composite resin restorations. In recent, the dual cure resin cement was developed with the advocations that it could increase the curing rates in the sites where the curing ligt could not reach. Moreover many manufactors added some adhesive components in the resin cement. This study was undertaken to observe the effects of curing depth and light curing times on the degree of conversion of dual cure resin cements. CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND, by the Fourier transform Infrared analysis, changing the curing depth 1mm, 2mm and 3mm, and varying the light curing time 20 seconds, 40 seconds and 80 seconds at each depth. The cytotoxicity of dual cure resin cements was tested by the in vitro MTT method using L929 cell. The results was evaluated and compared statistically. The results were obtained as follows : 1. The dual cure resin cements reavealed various degree of conversion, CR INLAY CEMENT and DUAL CEMENT had a tendency to be more reactive to the light cure and OPTEC BOND was a more chemical one. 2. CR INLAY CEMENT and DUAL CEMENT showed the lowest degree of conversion in 2 mm depth, and in 3mm depth the degree of conversion increased, which were due to the chemical cure of dual cures, but OPTEC BOND showed decreasing degree of conversion with increasing curing dept h and all experimental groups showed lower degree of conversion than CHEMICAL group which cured in dark room with no light, so the weak light-curing of dual cure resin cement prevented the chemical cure. (P<0.05) 3. CR INLAY CEMENT and DUAL CEMENT showed increasing degree of conversion in 1 mm and 3 mm, according to the increasing cure times, but in 2 mm depth the degree of conversion decreased with increasing light-curing times and OPTEC BOND showed contrary tendency, but there was no ststistical importance in the differences among the experimental group.(P>0.05) 4. The optical density by MTT assay of extractions of CR INLAY CEMENT, DUAL CEMENT and OPTEC BOND revealed no statitically important differences comparing with optical density of negative control.(P>0.05) 5. CR INLAY CEMENT showed a tendency of increaing cytotoxicity with days and DUAL CEMENT and OPTEC BOND showed higher cytotoxicity in 2 days than in 4 days, but there was no statistical importance in the differences.(P>0.05).

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DENTAL TREATMENT IN A PATIENT WITH METACHROMATIC LEUKODYSTROPHY UNDER GENERAL ANESTHESIA : A CASE REPORT (이염성 백질 이영양증 환아의 치아우식 치료 증례 보고)

  • Ryu, Jiyeon;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Young Jae;Kim, Jung-Wook;Jang, Ki-Taeg;Kim, Chong-Chul;Lee, Sang-Hoon
    • The Journal of Korea Assosiation for Disability and Oral Health
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    • v.12 no.2
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    • pp.96-100
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    • 2016
  • Metachromatic leukodystrophy (MLD) is a progressive and degenerative neurological disease caused by a deficiency of the catabolic enzyme arylsulfatase A. Deficiency of arylsulfatase A results in accumulation of sulfatide in the white matter of the peripheral and central nervous system and it occurs demyelination as a result. The patient gradually goes through mental and motor failure. General symptoms of MLD include gait disturbance, mental deterioration, muscle rigidity and impaired swallowing. Inheritance of the disease is autosomal recessive. We report a dental caries treatment of a 3-year old boy with MLD. The patient underwent hematopoietic stem cell transplantation (HSCT) to slow the progression of the disease. He was suffered from difficulties of mastication and swallowing from the degenerative neurological symptom. He was ingesting food by both oral feeding and tubal feeding after he took percutaneous endoscopic gastrostomy (PEG). The cause of multiple caries was mainly presumed as patient's prolonged time of meal. The treatment was performed under general anesthesia considering patient's incompliance. Severely affected lower primary molars were treated with pulp treatment and restored with stainless steel crown. Others were restored with composite resin. There were no postoperative complications. MLD is life threatening progressive disease and also has an impact on unfavorable condition for oral health. Routine home oral care and periodic professional dental care should be emphasized to the caregiver of patient considering the susceptibility of dental caries. Not only the medical care, but periodic dental office visit would benefit the quality of life of the patient.

The Analgesic Effects of Apitoxin and its Mechanism via JOR and Measuring Expression of mRNA in Phospholipase and TPH using RT-PCR (Jaw Opening Reflex 및 RT-PCR을 이용한 봉독의 진통효과)

  • Cho, Kwang-Ho;Lee, Jae-Dong;Park, Dong-Suk;Ahn, Byoung-Choul
    • Journal of Pharmacopuncture
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    • v.3 no.1
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    • pp.35-51
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    • 2000
  • The purpose of this study is to prove the analgesic effects of apitoxin and its mechanism via jaw-opening reflex(JOR) and measuring expression of mRNA in Phospholipase and Tryptophan hydroxylase(TPH) using RT-PCR. The experiments were carried out on Sprague-Dawley rats(300-400g) and mastocytoma(P-185 HTR) for JOR and RT-PCR, respectively. Rats anesthetized with thiopental sodium (80mg/kg) were used in the Tooth Pulp stimulation induced JOR. The amplitude of a digastric electromyogram (dEMG) was recorded during the stimulation at an intensity of 1.5 times the threshold for JOR. Apitoxin used in this experiment was diluted with normal saline by 1:1000. Apitoxin was injected intravenously into the test group while normal saline to the control group. However, it was injected directly into the cell of mastocytoma. We referred to base sequence registered in Genbank in designing primers for RT-PCR. The results were as follows; (1)Compared with control group, analgesic effect started to show right after Sprague-Dawely rats were treated with apitoxin($71.50{\pm}8.08$) and lasted for 50 minutes. (2)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of phospholipase or rate-limiting enzyme of biosynthesis of prostaglandins with $10{\mu}g/ml$ apitoxin.($31.74{\pm}18.98%$, P<0.05) (3)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of TPH or rate-limiting enzyme in biosynthesis of serotonin with $10{\mu}g/ml$ apitoxin.($131.37{\pm}16.87%$, P<0.05). These results suggest that $10{\mu}g/ml$ apitoxin have the most analgesic effects. This study showed that apitoxin has analgesic effects and held good for 50 minutes. The injection of apitoxin has brought out changes in the degree of expression of phospholipase and TPH. These results strongly suggest that analgesic mechanism by apitoxin is closely related to prostaglandins and serotonin.

The Expression of Type I Collagen in Periodontal Tissue during the Experimental Movement of Rat Incisors (백서의 실험적 치아 이동시 교원질 발현에 관한 면역조직화학적 연구)

  • Kim, Sang-Cheol;Jeon, In-Seop
    • The korean journal of orthodontics
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    • v.26 no.4
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    • pp.455-467
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    • 1996
  • This study was designed to evaluate the expression of type I collagen in periodontal tissue during the experimental movement of rat incisors. Twenty-one Sprague-Dawley rats were divided into a control group(3 rats), and experimental groups(18 rats) where a force(75g) from helical springs across the maxillary incisors was applied. Experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And tissue slides of control and experimental groups were studied histologically and immunohistochemically by LSAB(Labelled streptavidine Biotin) immunohistochemical staining for type I collagen. The results were as follows: 1. Until 28-day after force application, periodontal fibers were strectched on the tension side, and compressed in pressure side, and the arrangement of periodontal fibers was not recovered by that time. 2. The degree of type I collagen expression in control group was rare in the oral epithelium, predentin, pulp and periodontal ligament, but was mildly positive in osteoblasts, acellular cementum, cementoblasts, intermaxillary suture. 3. At acellular cementum of experimental group, the expression of type I collagen was moderate in 1-day and severe in 7-day, which was maintained until 28-day. 4. Type I collagen was observed in the newly formed fibrous connective tissue and osteoblasts at intermaxillary suture, moderately in 1-day, and severely in 14-day. 5. The tension side of periodontal ligament showed a more positive expression of type I collagen than the pressure side in 4-day. The degree was highest in 7-day and was not differentiated between sides in 14-day. 6. In the side wall of bone matrix on which osteoblasts were attached, type I collagen was expressed severely, especially in 7-day. From the above findings, we could suggest that bone remodeling in tooth movement be intimately related to the cell differentiation and the resulting formation of type I collagen.

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EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.

Bioleaching of Mn(II) from Manganese Nodules by Bacillus sp. MR2 (Bacillus sp. MR2에 의한 망간단괴의 생물용출)

  • Choi, Sung-Chan;Lee, Ga-Hwa;Lee, Hong-Keum
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.411-415
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    • 2009
  • Some microorganisms are capable of leaching Mn(II) from nonsulfidic manganese ores indirectly via nonenzymatic processes. Such reductive dissolution requires organic substrates, such as glucose, sucrose, or galactose, as a source of carbon and energy for microbial growth. This study investigated characteristics of Mn(II) leaching from manganese nodules by using heterotrophic Bacillus sp. strain MR2 provided with corn starch as a less-expensive substrate. Leaching of Mn(II) at 25.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$ was accompanied with cell growth, but part of the produced Mn(II) re-adsorbed onto residual $MnO_2$ particles after 24 h. Direct contact of cells to manganese nodule was not necessary as a separation between them with a dialysis tube produced similar amount [24.6 g Mn(II) $kg^{-1}$ nodule $day^{-1}$]. These results indicated an involvement of extracellular diffusible compound(s) during Mn(II) leaching by strain MR2. In order to optimize a leaching process we tested factors that influence the reaction, and the most efficient conditions were $25\sim35^{\circ}C$, pH 5~7, inoculum density of 1.5~2.5% (v/v), pulp density of 2~3 g/L, and particle size <75 ${\mu}m$. Although Mn(II) leaching was enhanced as particle size decrease, we suggest <212 ${\mu}m$ as a proper size range since more grinding means more energy consumption The results would help for the improvement of bioleaching of manganese nodule as a less expensive, energy-efficient, and environment-friendly technology as compared to the existing physicochemical metal recovery technologies.