• Title/Summary/Keyword: in vitro techniques

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Secondary Metabolites from Enzymatic Oxidation of Caffeic Acid with Pancreatic Lipase Inhibitory Activity (카페인산의 효소적 산화반응으로부터 췌장 지방분해효소 저해 물질의 분리)

  • Kim, Tae Hoon;Kim, Myoung Kwon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1912-1917
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    • 2015
  • Pancreatic lipase is a potential therapeutic target for the treatment of diet-induced obesity in humans. As part of our continuing search for novel bioactive compounds, the convenient enzymatic transformation of caffeic acid into neolignans as well as related oxidized-products enhanced pancreatic lipase inhibitory activity. Enzymatic transformation of caffeic acid (1) using polyphenol oxidase originating from Korean pear yielded four oxidized metabolites, which were identified by different spectroscopic techniques ($^1H$,$^{13}C$ NMR, DEP/T, $^1H-^1H$ COSY, HMBC, HMQC, and NOESY). The anti-obesity efficacy of caffeic acid reactant was tested by in vitro porcine pancreatic lipase assay. All tested samples showed dose-dependent pancreatic lipase inhibitory activities. Four oxidative products including phellinsin A (2), caffeicinic acid (3), isocaffeicinic acid (4), and 7,8-erythro-caffeicin (5) were isolated and identified. The major metabolites (2~5) were evaluated for their pancreatic lipase inhibitory activity, and oxidized-products (2~3) improved potency against pancreatic lipase when compared to original caffeic acid. This result suggested that the neolignans isolated from oxidative transformation of caffeic acid might be beneficial in the treatment of obesity and relevant diseases, and the convenient enzymatic transformation by polyphenol oxidase may be a valuable method for structural modification and enhancement of activity.

Fitting accuracy of ceramic veneered Co-Cr crowns produced by different manufacturing processes

  • von Maltzahn, Nadine Freifrau;Bernhard, Florian;Kohorst, Philipp
    • The Journal of Advanced Prosthodontics
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    • v.12 no.2
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    • pp.100-106
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    • 2020
  • PURPOSE. The purpose of this in vitro study was to evaluate the fitting accuracy of single crowns made from a novel presintered Co-Cr alloy prepared with a computer-aided design and computer-aided manufacturing (CAD/CAM) technique, as compared with crowns manufactured by other digital and the conventional casting technique. Additionally, the influence of oxide layer on the fitting accuracy of specimens was tested. MATERIALS AND METHODS. A total of 40 test specimens made from Co-Cr alloy were investigated according to the fitting accuracy using a replica technique. Four different methods processing different materials were used for the manufacture of the crown copings (milling of presintered (Ceramill Sintron-group_cer_sin) or rigid alloy (Tizian NEM-group_ti_nem), selective laser melting (Ceramill NPL-group_cer_npl), and casting (Girobond NB-group_gir_nb)). The specimens were adapted to a resin model and the outer surfaces were airborne-particle abraded with aluminum oxide. After the veneering process, the fitting accuracy (absolute marginal discrepancy and internal gap) was evaluated by the replica technique in 2 steps, before removing the oxide layer from the intaglio surface of the crowns, and after removing the layer with aluminum oxide airborne-particle abrasion. Statistical analysis was performed by multifactorial analysis of variance (ANOVA) (α=.05). RESULTS. Mean absolute marginal discrepancy ranged between 20 ㎛ (group_cer_npl for specimens of Ceramill NPL) and 43 ㎛ (group_cer_sin for crowns of Ceramill Sintron) with the oxide layer and between 19 ㎛ and 28 ㎛ without the oxide layer. The internal gap varied between 33 ㎛ (group_ti_nem for test samples of Tizian NEM) and 75 ㎛ (group_gir_nb for the base material Girobond NB) with the oxide layer and between 30 ㎛ and 76 ㎛ without the oxide layer. The absolute marginal discrepancy and the internal gap were significantly influenced by the fabrication method used (P<.05). CONCLUSION. Different manufacturing techniques had a significant influence on the fitting accuracy of single crowns made from Co-Cr alloys. However, all tested crowns showed a clinically acceptable absolute marginal discrepancy and internal gap with and without oxide layer and could be recommended under clinical considerations. Especially, the new system Ceramill Sintron showed acceptable values of fitting accuracy so it can be suggested in routine clinical work.

Implantable Flexible Sensor for Telemetrical Real-Time Blood Pressure Monitoring using Polymer/Metal Multilayer Processing Technique (폴리머/ 금속 다층 공정 기술을 이용한 실시간 혈압 모니터링을 위한 유연한 생체 삽입형 센서)

  • Lim Chang-Hyun;Kim Yong-Jun;Yoon Young-Ro;Yoon Hyoung-Ro;Shin Tae-Min
    • Journal of Biomedical Engineering Research
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    • v.25 no.6
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    • pp.599-604
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    • 2004
  • Implantable flexible sensor using polymer/metal multilayer processing technique for telemetrical real-time blood pressure monitoring is presented. The realized sensor is mechanically flexible, which can be less invasively implanted and attached on the outside of blood vessel to monitor the variation of blood pressure. Therefore, unlike conventional detecting methods which install sensor on the inside of vessel, the suggested monitoring method can monitor the relative blood pressure without injuring blood vessel. The major factor of sudden death of adults is a disease of artery like angina pectoris and myocardial infarction. A disease of circulatory system resulted from vessel occlusion by plaque can be preventable and treatable early through continuous blood pressure monitoring. The procedure of suggested new method for monitoring variation of blood pressure is as follows. First, integrated sensor is attached to the outer wall of blood vessel. Second, it detects mechanical contraction and expansion of blood vessel. And then, reader antenna recognizes it using telemetrical method as the relative variation of blood pressure. There are not any active devices in the sensor system; therefore, the transmission of energy and signal depends on the principle of mutual inductance between internal antenna of LC resonator and external antenna of reader. To confirm the feasibility of the sensing mechanism, in vitro experiment using silicone rubber tubing and blood is practiced. First of all, pressure is applied to the silicone tubing which is filled by blood. Then the shift of resonant frequency with the change of applied pressure is measured. The frequency of 2.4 MHz is varied while the applied pressure is changed from 0 to 213.3 kPa. Therefore, the sensitivity of implantable blood pressure is 11.25 kHz/kPa.

Effect of Porcelain Firing Process on the Marginal and Internal Fit of Ni-Cr Alloy Metal-Ceramic Crown (도재 소성과정이 Ni-Cr 금속도재관의 변연 및 내면 적합도에 미치는 영향)

  • Kim, Ki-Baek;Kim, Seok-Hwan;Kim, Jae-Hong
    • Journal of dental hygiene science
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    • v.14 no.3
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    • pp.405-410
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    • 2014
  • The purpose of this study in vitro investigation was to compare the marginal and internal fit of Ni-Cr alloy metal ceramic crown before and after porcelain veneering. Furthermore, this study evaluated whether the influence of the porcelain firing on the precision of fit of dental prostheses. The maxillary right incisor was selected as an abutment for experiments. Ten working models were prepared. Ni-Cr alloy cores appropriate for each abutment were prepared by lost wax technique. The marginal area and four internal areas of the crowns were measured at two stages: before veneering process and after upper porcelain firing. Silicone replica techniques were used. The data were statistically analyzed with the paired t-test (${\alpha}=0.05$). $Mean{\pm}SD$ marginal and internal gap were $67.1{\pm}23.3{\mu}m$ for the nickle chrome alloy core group and $74.4{\pm}21.9{\mu}m$ for the metal ceramic crown group. There were statistically significant differences in all investigated areas (p<0.05). Within the limitations of this study, none of the Ni-Cr alloy metal crown values measured after porcelain firing process exceeded $120{\mu}m$, which is the clinically acceptable threshold.

Comparison of in vitro Antimicrobial Activities of Tc-99m Infecton and Ciprofloxacin (Tc-99m Infecton과 Ciprofloxacin의 생체외 항균력 비교)

  • Kim, Sung-Min;Bom, Hee-Seung;Song, Ho-Chun;Jeong, Hwan-Jeong;Kim, Ji-Yeul;Shin, Jong-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.35 no.1
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    • pp.75-80
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    • 2001
  • Purpose: There was little evidence that Tc-99m labeled ciprofloxacin (Infecton) located inside of bacteria. Antimicrobial activity of Infecton could be an indirect evidence of its location. We compared in vitro antimicrobial activities of Infecton and ciprofloxacin. Materials and methods: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Infecton and ciprofloxacin against three standard strains of bacteria, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were measured using modified broth macrodilution techniques and pour plate methods, respectively. Data were expressed as mean${\pm}$SE (range). Results: MICs of Infecton and ciprofloxacin were $1.12{\pm}0.20\;(0.8{\sim}1.6){\mu}g/ml\;and\;0.35{\pm}0.04\;(0.2{\sim}0.4){\mu}g/ml$ for S. aureus, $0.03{\pm}0.005\;(0.025{\sim}0.05){\mu}g/ml\;and\;0.011{\pm}0.001\;(0.006{\sim}0.012){\mu}g/ml$ for E. coil, and $0.96{\pm}0.16\;(0.8{\sim}1.6){\mu}g/ml)\;and\;0.56{\pm}0.098\;(0.4{\sim}0.8){\mu}g/ml$ for P. aeruginosa, respectively. MBCs of Infecton and ciprofloxacin were $2.56{\pm}0.39\;(1.6{\sim}3.2){\mu}g/ml\;and\;0.88{\pm}0.2\;(0.4{\sim}1.6){\mu}g/ml$ for S. aureus, $0.04{\pm}0.05\;(0.025{\pm}0.05){\mu}g/ml\;and\;0.02{\pm}0.01\;(0.025{\sim}0.05)\;{\sim}g/ml$ for E coli, and $2.24{\pm}0.39\;(1.6{\sim}3.2){\mu}g/ml\;and\;1.44{\pm}0.16\;(0.8{\sim}1.6){\mu}g/ml$ for P. aeruginosa, respectively. Conclusion: Although both MICs and UBCs of Infecton were higher than those of ciprofloxacin, all three standard bacterial strains were sensitive to Infecton. It could be an indirect evidence that Tc-99m Infecton be a specific imaging agent for bacterial infection.

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In vitro Antitumor Activity of Ergosterol Peroxide Isolated from Cordyceps militaris on Cancer Cell Lines from Korean Patients (Cordyceps militaris로부터 분리한 Ergosterol Peroxide의 한국인 암세포주에 대한 항암작용)

  • Kim, Ha-Won;Kim, Young-Ho;Cai, Xing Fu;Nam, Kyong-Suk;Lee, Seung-Jung;An, Hye-Suk;Jeong, Eun-Ho;Yun, Seung-Hyun;Sung, Su-Kyong;Lee, Sung-Jin;Hyun, Jin-Won
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.61-66
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    • 2001
  • Entomopathogenic fungus of Cordyceps militaris is famous of its medicinal efficacies. An antitumor compound was purified from the n-hexane extract of artificially cultivated fruiting bodies of C. militaris and identified as ergosterol peroxide $(5{\alpha},\;8{\alpha}-epidioxy-24(R)-methylcholesta-6,22-dien-3{\beta}-o1,\; C_{28}H_{44}O_3)$ mainly by $^1H\;and\;^{13}C-NMR$ spectroscopic techniques. When the antitumor activity of ergosterol peroxide was measured against 3 tumor cell lines from Korean cancer patients, it showed the most strong activity against gastric cancer SNU-l cell line 3 days after treatment. The 50% inhibitory concentrations $(IC_{50})$ of ergosterol peroxide 6 days after treatment were $75.8{\mu}g/ml$ for human gastric SNU-1 tumor cell line, $39.7{\mu}g/ml$ for human colorectal SNU-C4 tumor cell line and $32.7{\mu}g/ml$ for human hepatoma SNU-354 cell line.

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Steroidal Saponins from the Rhizomes of Asparagus oligoclonos and their Antibacterial Activity (방울비짜루(Asparagus oligoclonos)로부터 분리한 스테로이드 사포닌의 항균활성)

  • Seong, Jae-Duck;Park, Seung-Yong;Oh, Sei-Ryang;Kwack, Yong-Ho;Kim, Geum-Soog
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.136-140
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    • 2000
  • Two steroidal saponins were isolated from MeOH extract of the rhizomes of Asparagus oligoclonos. Using the spectroscopic techniques of $^1H,\;^{13}C$ NMR and 2D NMR, they were identified as $3-O-[{\beta}-D-glucopyranosyl-(1{\rightarrow}2)-{\beta}-D-glucopyranosyl]-(25S)-spirostan-3{\beta}-ol$ and $3-O-{{\beta}-D-glucopyranosyl-(1{\rightarrow}2)-[{\beta}-D-xylopyranosyl-(1{\rightarrow}4)]-{\beta}-D-glucopyranosyl}-(25S)-spirostan-3{\beta}-ol$, respectively. They were first isolated from A. oligoclonos. The in vitro antibacterial activities as well as antibacterial spectrums against 20 bacteria strains were investigated with MIC test. Both saponins inhibited the growth of 10 bacteria strains at the concentration of$100\;{\mu}g/ml$.

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Mass Propagation of Hypolepis punctata (Thunb.) Mett. Using In Vitro Culture Techniques (조직배양을 이용한 점고사리의 대량증식 방법)

  • Park, Kyungtae;Jang, Bo Kook;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.57-57
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    • 2019
  • 본 연구는 관상 및 조경용으로 개발이 가능한 남방계 양치식물인 점고사리[Hypolepis punctata (Thunb.) Mett.]의 전엽체 증식 및 포자체 형성에 적합한 배양조건을 구명하고자 수행되었다. 무가온 온실에서 성숙한 포자엽을 채집한 후 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 8주 간격으로 계대배양 하여 실험의 재료로 사용하였다. 배지종류에 따른 전엽체의 증식 및 형태형성을 확인하고자, 배양된 전엽체 0.3g을 메스로 잘게 다진 후, 농도를 1/4, 1/2, 1, 2배로 조절한 MS배지에 8주간 배양하였다. 이후 선발된 배지를 기준으로 sucrose, 활성탄, 질소급원의 농도를 조절하여 전엽체의 증식과 형태형성을 확인하였다. 그 결과, 1MS배지에서 전엽체의 생체중이 초기 접종량인 0.3g에 비해 10.7배 증가한 3.2g으로 가장 높은 증가율을 보였다. 형태관찰에서도 장정기의 형성이 관찰되었으며, 전엽체의 쿠션조직이 비교적 잘 발달하였다. 전엽체 증식에 가장 좋은 효과를 보인 1MS배지를 기준으로 sucrose의 농도를 0-4%로 달리하여 실험한 결과, 1%의 처리구에서 6.7g으로 가장 높은 생체중을 보였다. 활성탄의 농도를 0-0.8%로 첨가한 네 처리구 중에서는 0.8%의 처리구에서 14.2g으로 무처리구에 비해 생체중이 2배 이상 증가하였다. 전엽체의 형태 또한 정상적인 발달을 보였다. 질소급원의 비율을 30-120mM로 조절한 배지에서는 60mM의 처리구에서 4.9g으로 가장 높은 생체중을보였다. 이후 포자체 형성을 위한 최적의 토양조건을 구명하고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 달리하여 5종류의 배양토를 조성하였다. 혼합된 토양은 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진 하였으며, 배양된 전엽체 1g을 증류수와 함께 10초간 분쇄한 다음 준비된 토양표면에 분주하여 재배하였다. 12주간의 재배 결과, 원예상토를 단용한 토양에서 포자체의 수가 포트당 250.0개로 가장 많이 형성되었으며, 포자체의 생육 또한 다른 처리구에 비해 우수한 결과를 보였다. 한편 피트모스가 혼합된 토양에서는 포자체가 형성되지 않았다. 따라서 점고사리의 전엽체 대량증식에 적합한 배지는 sucrose 1%와 질소급원의 농도를 60mM, 활성탄을 0.8% 첨가한 1MS배지로 판단되며, 포자체 대량생산을 위해서는 원예상토를 단용한 토양이 적합하다고 판단된다.

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Microassisted Fertilization of Human Oocytes with Intracytoplasmic Sperm Injection in IVF-ET Patients with History of Failure in Fertilization or Extremely Low Fertilization Rate in Previous Cycles (미수정 및 저수정율의 기왕력을 지닌 체외수정시술 환자에서의 난자 세포질내 정자 주입술을 이용한 미세보조 수정술에 관한 연구)

  • Moon, Shin-Yong;Kim, Seok-Hyun;Chae, Hee-Dong;Kim, Kwang-Rye;Lee, Jae-Hoon;Kim, Hee-Sun;Ryu, Buom-Yong;Oh, Sun-Kyung;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.83-93
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    • 1997
  • Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization (MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after partial zona dissection (PZD), subzonal insertion (SUZI), and intracytoplasmic sperm injection (ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not undergo standard IVF-ET repetitively because of failure in fertilization or extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles. From March, 1995 to May, 1996, 27 cycles of IVF-ET with ICSI in 19 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $10.50{\pm}6.13$ in 30 previous cycles, and $10.57{\pm}5.53$ in 27 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was $7.89{\pm}4.30$, and the fertilization rate of $67.9{\pm}20.2%$ could be obtained after ICSI. The number of embryos transferred was $1.43{\pm}2.40$ in previous cycles, and $4.36{\pm}1.77$ with the mean CES of $41.8{\pm}27.4$ in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 29.6% (8/27) per cycle and 42.1% (8/19) per patient with the clinical pregnancy rate of 22.2% (6/27) per cycle and 31.6% (6/19) per patient. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with the past history of failure in fertilization or low fertilization rate of oocytes in the previous IVF-ET cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to effective management of infertile couples.

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A STUDY ON THE MEASUREMENT OF THE IMPLANT STABILITY USING RESONANCE FREQUENCY ANALYSIS (공진 주파수 분석법에 의한 임플랜트의 안정성 측정에 관한 연구)

  • Park Cheol;Lim Ju-Hwan;Cho In-Ho;Lim Heon-Song
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.2
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    • pp.182-206
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    • 2003
  • Statement of problem : Successful osseointegration of endosseous threaded implants is dependent on many factors. These may include the surface characteristics and gross geometry of implants, the quality and quantity of bone where implants are placed, and the magnitude and direction of stress in functional occlusion. Therefore clinical quantitative measurement of primary stability at placement and functional state of implant may play a role in prediction of possible clinical symptoms and the renovation of implant geometry, types and surface characteristic according to each patients conditions. Ultimately, it may increase success rate of implants. Purpose : Many available non-invasive techniques used for the clinical measurement of implant stability and osseointegration include percussion, radiography, the $Periotest^{(R)}$, Dental Fine $Tester^{(R)}$ and so on. There is, however, relatively little research undertaken to standardize quantitative measurement of stability of implant and osseointegration due to the various clinical applications performed by each individual operator. Therefore, in order to develop non-invasive experimental method to measure stability of implant quantitatively, the resonance frequency analyzer to measure the natural frequency of specific substance was developed in the procedure of this study. Material & method : To test the stability of the resonance frequency analyzer developed in this study, following methods and materials were used : 1) In-vitro study: the implant was placed in both epoxy resin of which physical properties are similar to the bone stiffness of human and fresh cow rib bone specimen. Then the resonance frequency values of them were measured and analyzed. In an attempt to test the reliability of the data gathered with the resonance frequency analyzer, comparative analysis with the data from the Periotest was conducted. 2) In-vivo study: the implants were inserted into the tibiae of 10 New Zealand rabbits and the resonance frequency value of them with connected abutments at healing time are measured immediately after insertion and gauged every 4 weeks for 16 weeks. Results : Results from these studies were such as follows : The same length implants placed in Hot Melt showed the repetitive resonance frequency values. As the length of abutment increased, the resonance frequency value changed significantly (p<0.01). As the thickness of transducer increased in order of 0.5, 1.0 and 2.0 mm, the resonance frequency value significantly increased (p<0.05). The implants placed in PL-2 and epoxy resin with different exposure degree resulted in the increase of resonance frequency value as the exposure degree of implants and the length of abutment decreased. In comparative experiment based on physical properties, as the thickness of transducer increased, the resonance frequency value increased significantly(p<0.01). As the stiffness of substances where implants were placed increased, and the effective length of implants decreased, the resonance frequencies value increased significantly (p<0.05). In the experiment with cow rib bone specimen, the increase of the length of abutment resulted in significant difference between the results from resonance frequency analyzer and the $Periotest^{(R)}$. There was no difference with significant meaning in the comparison based on the direction of measurement between the resonance frequency value and the $Periotest^{(R)}$ value (p<0.05). In-vivo experiment resulted in repetitive patternes of resonance frequency. As the time elapsed, the resonance frequency value increased significantly with the exception of 4th and 8th week (p<0.05). Conclusion : The development of resonance frequency analyzer is an attempt to standardize the quantitative measurement of stability of implant and osseointegration and compensate for the reliability of data from other non-invasive measuring devices It is considered that further research is needed to improve the efficiency of clinical application of resonance frequency analyzer. In addition, further investigation is warranted on the standardized quantitative analysis of the stability of implant.