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Evaluation of Protective Immune Response Induced by a DNA Vaccine Encoding GRA8 against Acute Toxoplasmosis in a Murine Model

  • Chu, Jia-Qi;Huang, Shuai;Ye, Wei;Fan, Xuan-Yan;Huang, Rui;Ye, Shi-Cai;Yu, Cai-Yuan;Wu, Wei-Yun;Zhou, Yu;Zhou, Wei;Lee, Young-Ha;Quan, Juan-Hua
    • The Korean Journal of Parasitology
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    • v.56 no.4
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    • pp.325-334
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    • 2018
  • Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), $IFN-{\gamma}$, $TNF-{\alpha}$, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.

Effects of enamel matrix derivative and titanium on the proliferation and differentiation of osteoblasts (법랑기질유도체를 도포한 타이태늄 표면에서 조골세포의 증식 및 분화)

  • Park, Sang-Hyun;Lee, In-Kyeong;Yang, Seung-Min;Shin, Seung-Yun;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.33 no.3
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    • pp.359-372
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    • 2003
  • Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

Preparation and Characterization of Crosslinked Copolymer Membrane Containing Sulfonated Poly(ether sulfone) and p-Phenylene Terephthalamide Segments (Sulfonated Poly(ether sulfone)과 p-Phenylene Terephthalamide 세그먼트를 포함하는 가교 공중합체 멤브레인의 제조 및 특성 연구)

  • Kim, Jung-Min;Hwang, Seung-Sik;Cho, Chang-Gi
    • Polymer(Korea)
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    • v.35 no.2
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    • pp.106-112
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    • 2011
  • Aromatic copolyamides were prepared and their applicability to proton exchange membrane was studied. The copolymers contain two segments; thermally stable and mechanically strong poly (p-phenylene terephthalamide) (PPTA), and easily processable and good film-forming polysulfone. For the copolymers, different ratios of amine-terminated sulfonated ether sulfone monomer, terephthaloyl chloride, and p-phenylene diamine were sequentially reacted. The obtained copolymers were mixed with trimethylolpropane triglycidyl ether (TMPTGE), thermally cured, and converted into proton exchange membranes for fuel cell application. The reactions at each step and the molecular characteristics of precursor copolymers were confirmed by $^1H$ NMR, FTIR, and titration. The performance of the membranes was measured in terms of water uptake and proton conductivity. The water uptake, ion exchange capacity (IEC), and proton conductivity of the membranes increased with the increase of sulfonated ether sulfone segment content. Membrane containing 60 mol% sulfonic acid sulfone segment showed 1.88 meq/g IEC value. Water uptake was limited less than 110 wt% and the highest proton conductivity was up to $7.4{\times}10^{-2}$ S/cm ($25^{\circ}C$, RH=100%).

Synthesis of Crosslinkable m-Aramid Ionomer Containing Sulfonated Ether Sulfone and Their Characterization for PEMFC Membrane (Sulfonated Ether Sulfone을 포함한 Crosslinkable m-Aramid계 Ionomer의 합성과 연료전지 막으로의 이용)

  • Jung, Hyun-Jin;Kim, Jung-Min;Cho, Chang-Gi
    • Polymer(Korea)
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    • v.34 no.3
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    • pp.202-209
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    • 2010
  • Aromatic copolyamides were prepared and their applicability to proton exchange membrane wasstudied. The copolymer contains thermally stable and mechanically strong poly(m-phenylene isophthalamide) segments, and easily processable and good film forming polysulfone segments. For the copolymer, amineterminated sulfonated ether sulfone monomer, m-phenylene diamine, and isophthaloyl chloride were reacted, and the obtained copolymer was transformed into crosslinkable prepolymer by the reaction with acryloyl chloride. The prepolymer was thermally cured and converted into proton exchange membranes for fuel cell application. Each reaction step and the molecular characteristics of precursor copolymers were monitored and confirmed by $^1H$ NMR, FTIR, and titration. The performance of the membranes was measured in terms of water uptake, proton conductivity, and thermal stability. The water uptake, ion exchange capacity (IEC), and proton conductivity of the membranes increased with the increase of sulfonated ether sulfone segment content. Membrane containing 30 mol% sulfonic acid sulfone segment showed 1.57 meq/g IEC value. Water uptake was limited less than 44 wt% and the highest proton conductivity up to $3.93{\times}10^{-2}S/cm$ ($25^{\circ}C$, RH= 100%) was observed.

Analysis of Greenhouse Thermal Environment by Model Simulation (시뮬레이션 모형에 의한 온실의 열환경 분석)

  • 서원명;윤용철
    • Protected Horticulture and Plant Factory
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    • v.5 no.2
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    • pp.215-235
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    • 1996
  • The thermal analysis by mathematical model simulation makes it possible to reasonably predict heating and/or cooling requirements of certain greenhouses located under various geographical and climatic environment. It is another advantages of model simulation technique to be able to make it possible to select appropriate heating system, to set up energy utilization strategy, to schedule seasonal crop pattern, as well as to determine new greenhouse ranges. In this study, the control pattern for greenhouse microclimate is categorized as cooling and heating. Dynamic model was adopted to simulate heating requirements and/or energy conservation effectiveness such as energy saving by night-time thermal curtain, estimation of Heating Degree-Hours(HDH), long time prediction of greenhouse thermal behavior, etc. On the other hand, the cooling effects of ventilation, shading, and pad ||||&|||| fan system were partly analyzed by static model. By the experimental work with small size model greenhouse of 1.2m$\times$2.4m, it was found that cooling the greenhouse by spraying cold water directly on greenhouse cover surface or by recirculating cold water through heat exchangers would be effective in greenhouse summer cooling. The mathematical model developed for greenhouse model simulation is highly applicable because it can reflects various climatic factors like temperature, humidity, beam and diffuse solar radiation, wind velocity, etc. This model was closely verified by various weather data obtained through long period greenhouse experiment. Most of the materials relating with greenhouse heating or cooling components were obtained from model greenhouse simulated mathematically by using typical year(1987) data of Jinju Gyeongnam. But some of the materials relating with greenhouse cooling was obtained by performing model experiments which include analyzing cooling effect of water sprayed directly on greenhouse roof surface. The results are summarized as follows : 1. The heating requirements of model greenhouse were highly related with the minimum temperature set for given greenhouse. The setting temperature at night-time is much more influential on heating energy requirement than that at day-time. Therefore It is highly recommended that night- time setting temperature should be carefully determined and controlled. 2. The HDH data obtained by conventional method were estimated on the basis of considerably long term average weather temperature together with the standard base temperature(usually 18.3$^{\circ}C$). This kind of data can merely be used as a relative comparison criteria about heating load, but is not applicable in the calculation of greenhouse heating requirements because of the limited consideration of climatic factors and inappropriate base temperature. By comparing the HDM data with the results of simulation, it is found that the heating system design by HDH data will probably overshoot the actual heating requirement. 3. The energy saving effect of night-time thermal curtain as well as estimated heating requirement is found to be sensitively related with weather condition: Thermal curtain adopted for simulation showed high effectiveness in energy saving which amounts to more than 50% of annual heating requirement. 4. The ventilation performances doting warm seasons are mainly influenced by air exchange rate even though there are some variations depending on greenhouse structural difference, weather and cropping conditions. For air exchanges above 1 volume per minute, the reduction rate of temperature rise on both types of considered greenhouse becomes modest with the additional increase of ventilation capacity. Therefore the desirable ventilation capacity is assumed to be 1 air change per minute, which is the recommended ventilation rate in common greenhouse. 5. In glass covered greenhouse with full production, under clear weather of 50% RH, and continuous 1 air change per minute, the temperature drop in 50% shaded greenhouse and pad & fan systemed greenhouse is 2.6$^{\circ}C$ and.6.1$^{\circ}C$ respectively. The temperature in control greenhouse under continuous air change at this time was 36.6$^{\circ}C$ which was 5.3$^{\circ}C$ above ambient temperature. As a result the greenhouse temperature can be maintained 3$^{\circ}C$ below ambient temperature. But when RH is 80%, it was impossible to drop greenhouse temperature below ambient temperature because possible temperature reduction by pad ||||&|||| fan system at this time is not more than 2.4$^{\circ}C$. 6. During 3 months of hot summer season if the greenhouse is assumed to be cooled only when greenhouse temperature rise above 27$^{\circ}C$, the relationship between RH of ambient air and greenhouse temperature drop($\Delta$T) was formulated as follows : $\Delta$T= -0.077RH+7.7 7. Time dependent cooling effects performed by operation of each or combination of ventilation, 50% shading, pad & fan of 80% efficiency, were continuously predicted for one typical summer day long. When the greenhouse was cooled only by 1 air change per minute, greenhouse air temperature was 5$^{\circ}C$ above outdoor temperature. Either method alone can not drop greenhouse air temperature below outdoor temperature even under the fully cropped situations. But when both systems were operated together, greenhouse air temperature can be controlled to about 2.0-2.3$^{\circ}C$ below ambient temperature. 8. When the cool water of 6.5-8.5$^{\circ}C$ was sprayed on greenhouse roof surface with the water flow rate of 1.3 liter/min per unit greenhouse floor area, greenhouse air temperature could be dropped down to 16.5-18.$0^{\circ}C$, whlch is about 1$0^{\circ}C$ below the ambient temperature of 26.5-28.$0^{\circ}C$ at that time. The most important thing in cooling greenhouse air effectively with water spray may be obtaining plenty of cool water source like ground water itself or cold water produced by heat-pump. Future work is focused on not only analyzing the feasibility of heat pump operation but also finding the relationships between greenhouse air temperature(T$_{g}$ ), spraying water temperature(T$_{w}$ ), water flow rate(Q), and ambient temperature(T$_{o}$).

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Cooling Performance Deficiency of Air Conditioning System According to Air Quantity Included in Refrigerant (냉매 내 공기혼입에 따른 에어컨 시스템의 냉각성능 저하)

  • Moon, Seong-Won;Min, Young-Bong;Chung, Tae-Sang
    • Journal of Biosystems Engineering
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    • v.34 no.6
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    • pp.470-475
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    • 2009
  • This study was performed to present the diagnosis basis of cooling performance deficiency according to air quantity included in refrigerant of air-conditioner by detecting the temperatures and pressures of refrigerant pipeline. The car air-conditioner of SONATA III (Hyundai motor Co., Korea) was tested by maximum cooling condition at 1500 rpm of engine speed in the room with controlled air condition at $33\sim35^{\circ}C$ and 55~57% RH. Measured variables were temperature differences between inlet and outlet pipe surface of the compressor (Tcom), condenser (Tcon), receive dryer (Trec) and evaporator (Teva), and high pressure (HP) and low pressure (LP) in the refrigerant pipeline, and temperature difference (Tcoo) between inlet and outlet air of the cooling vent of evaporator. Control variables were the refrigerant charging weight and the vacuum degree in the refrigerant pipeline before charging refrigerant. From the test, it was represented that the measuring values of (Tcom), LP and (Tcoo) were enabled to make the diagnosis of cooling performance deficiency according to quantity included in refrigerant of air-conditioner. The ranges of Tcom, LP and Tcoo to make the diagnosis of cooling performance deficiency were respectively less than $55^{\circ}C$, more than 166.7 kPa-g(1.7 kgf/$cm^2$) and less than $13.7^{\circ}C$. In the case of using only external sensors and the condition under the normal performances of air conditioner, it was considered that the ranges of LP and Tcoo to make the diagnosis of cooling performance deficiency were respectively more than 166.7 Pa and less than $12^{\circ}C$.

Effects of Priming Progesterone on the LH Surge Expressions in Ovariectomized Shiba Goats (LH surge 발현에 대한 서로 다른 Progesterone 농도의 효과)

  • Kim, Seung-Joon
    • Journal of Veterinary Clinics
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    • v.31 no.1
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    • pp.25-30
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    • 2014
  • This study tested the hypothesis that the priming effects of progesterone on the timing of the LH surge induced by exogenous estradiol are more potentiated the negative feedback actions of progesterone on LH secretion by the existence of estradiol. In previous studies, the time interval from estradiol infusion until the peak of LH surge was gradually and significantly extended by the different levels of progesterone treated before estradiol infusions. Longterm ovariectomized Shiba goats that had received implants of estradiol capsules (Day 0) and three different progesterone silastic packet inducing follicular, subluteal and luteal levels of progesterone were divided into three groups such as non-P, low-P and high-P group. Blood samples were collected daily throughout the experiment for the analysis of gonadal steroid hormone levels. On Day 7, all devices of progesterone and estradiol packets were removed but estradiol capsules were maintained during the experiment, and blood samples were collected at 1 hr interval for 12 h from the time of progesterone removals to determine peripheral changes of estradiol and progesterone concentration. Then all animals were infused estradiol on the Day 7 after 13 h from the removals of progesterone devices with a peristaltic pump into jugular vein at a rate of 3-6 ${\mu}g/h$ for 36 h. For analysis of peripheral LH and estradiol concentration, blood samples were collected via another jugular vein at 2 h intervals for 52 h (from 4 h before the start of estradiol infusion to 48 h after the start of estradiol infusion). In all animals of the three groups treated with estradiol infusion, an LH surge was expressed but the peak time of LH surge was different. This time interval was not extended by the different levels of progesterone treated before estradiol infusions and the difference was not significant during this interval between the Low P and the High P groups. Progesterone pretreatment may contribute to regulating the neural system that is responded by estradiol, and estradiol existence potentiates the negative feedback effect of progesterone on GnRH/LH surge-generating system.

Development of antigen for the microplate latex agglutination test on toxoplasmosis in animals (Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단액 개발에 관한 연구)

  • Suh, Myung-deuk;Lee, Eung-goo
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.623-632
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    • 1993
  • This study was conducted to develop a sensitized latex-antigen for serodiagnosis of toxoplasmosis in animals. Tachyzoites of T gondii(RH-strain) harvested from mouse peritoneal cavity were purified through the filtraton of polycarbonate membrane(pore size, $3.0{{\mu}m}$, Costar Co.) and disrupted by ultrasonicator. The tachyzoite suspension was ultracentrifuged for 30 min at $60,000{\times}g(4{^{\circ}C})$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $0.8{{\mu}m}$ in diameter(Sigma) were used for the preparation of sensitized latex-antigen suspension. The several parameters including the preparation conditions, incubation buffer. serum dilution buffer and stability of agglutination reactions were evaluated and the results obtained were summarized as follows : 1. The antigen consisting of a water-lysate of T gondii tachyzoites was adsorbed onto polystyrene latex particles of $0.8{{\mu}m}$ in diameter by adding a latex suspension to an equal volume of diluted antigen solution and by incubating the mixture at $37{^{\circ}C}$ under different conditions. 2. The optimum incubation buffer used for the antigen sensitization was 0.1M Tris-HCl buffer(pH 8.0). 3. The optimum serum dilution buffer used for the latex agglutination test was 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300 mM NaCl. But 0.1M Tris-HCl-NaCl buffer(pH 7.4) containing 300-600 mM NaCl, 0.5% BSA and 0.01% Tween-20 improved the agglutination pattems and cleared the background of microplate well without the effects on L.A titer. 4. The time required for antigen sensitization was 40 and 60 min in incubation buffer(pH 8.0) at $37{^{\circ}C}$. But the optimun time for antigen sensitization was min at $37{^{\circ}C}$. 5. The optimun quantity of antigen absorbed on latex particles for proper agglutination was the range of 20 to $32{\mu}g$ of latex particles. 6. The optimun concentration of the latex-antigen suspension for the proper agglutination reaction was determined as 0.2%(w/v). 7. The specificity, rapidity and simplicity of the latex-particle agglutination test suggested that it might be adaptable to large scale serum screening.

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Optimization of Ingredients Formulation in tow Grades Surimi for Improvement of Gel Strength (저급 수리미의 젤 강도 증강을 위한 첨가물의 최적화)

  • CHOI Young-Joon;LEE Ho-Soo;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.5
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    • pp.556-562
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    • 1999
  • The increasing price of surimi has affected the economical benefits of surimi based food industry, To maintain gel strength in low grade surimi, the optimum formulation adding functional proteins to low grade surimi is required. The objective of this study was to develop the optimum formulation of ingredients in making gels in low grade surimi on the addition of functional non-muscle proteins to low grade surmi by measuring rheological properties of the gels. The rheological qualities of the cooked gels made with A and RA grade surimi on the effects of adding five kinds of starches (potato, wheat, waxy maize, corn and modified corn) and four kinds of functional proteins (bovine plasma protein, dehydrated egg white, soy protein isolate and whey protein concentrate) to the gels were evaluated, The gel styengths at cooking with A and RA grade surimi were decreased with increasing the added starches. The kind of starches added affected little the gel strengths in Rh grade surimi, while potato and corn starches decreased at the least in gel strengths of the gel made with A grade surimi with increasing the concentration of starches. The bovine plasma protein (BPP) significantly increased the gel strength, especially in RA grade surimi, but BPP decreased the whiteness of the gel. Therefore, the optimum content of BPP was up to $2\%$ because of the whiteness of the gels in RA grade surimi, The optimum formulation for the gel with RA grade surimi to satisfy the gel strength of 1000$\times$g and $78\%$ moisture was $40.9\%$ surimi, $9.1\%$ dehydrated egg white (DEW) and $0.9\%$ starch, while that with A grade surimi under the same condition was $37.9\%$ surimi, $6.6\%$ DEW and $3,4\%$ starch.

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Effects of $TGF-{\beta}1$ on Cellular Activity of Minocycline-Pretreated Human Periodontal Ligament Cells (($TGF-{\beta}$)이 Minocycline을 전처리한 사람 치주인대세포의 활성에 미치는 영향)

  • Yang, Seung-Oh;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.469-490
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    • 1996
  • The initial events required for periodontal regeneration is the attachment, spreading, and proliferation of appropriated cells at the healing sites. These have been reported that minocycline stimulates the attachment of periodontal ligament cells, and also $TGF-{\beta}1$ enhances the proliferation of periodontal ligament cells. The purpose of the present study was to evaluate the effects of $TGF-{\beta}1$ on the cellular activity of minocycline treated human periodontal ligament cells. Periodontal ligament cells were obtained from the explants of healthy periodontal ligaments of extracted 3rd molars or premolar teeth extracted from the patients for orthodontic treatment. The cells were cultured in minimal essential medium(${\alpha}-MEM$) supplemented with 10.000units/ml penicillin, $10,000{\mu}g/ml$ streptomycin and 10% FBS(fetal bovine serum) at $37^{\circ}C$ in a humidified atmosphere of 5% carbon dioxide and the 5th to the 8th passages of the cells were used. To evaluate the effect of minocycline on cell attachment, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After trypsinization, the cells were counted with hemocytometer and were taken photographs for observation of cellular morphology. To evaluate the effect of $TGF-{\beta}1$ on minocycline-pretreated periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4$ cells/ well in 24-well culture plates and treated with $20{\mu}g/ml$ and $100{\mu}g/ml$ of minocycline for 1.5 h. After incubation, 1 and 10ng/ml of $rh-TGF-{\beta}1$ were also added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay, DNA synthesis($^3H-thymidine\;assay$), and protein and collagen assay(3H-proline assay) were carried out. In the MIT assay, after 200ul MTT solutionlconeentration of 5mg/ml) were added to the each well of the 24-well plates and incubated for 3 hours, and 200 ul DMSO were added so as to dissolve insoluble blue formazan crystals which was formed in incubated period. Then it read plates on a ELISA reader. For mitogenic assay, 1 uCi/ml $^3H-thymidine$ was added to each well for the final 2 hours of the incubation periods. After labeling, the wells were washed 3 times with ice cold PBS and 4 times with 5% TCA to remove unincorporated label and precipitate the cellular DNA. DNA, with the incorporated $^3H-thymidine$, was solubilized with 500 ul of 0.1% NaOH/0.1% SDS. A 250 ul aliquot was removed from each well and placed in a scintillation vial with 4ml of scintillation cocktail. Using an liguid scintillation counter, counts per minute(CPM) were determined for each samples. 3 uCi/ml $^3H-proline$ was added to each well for the final 4 hours of the incubation periods and total protein and percent collagen synthesis were carried out. The results indicate that minocycline treated group with $100{\mu}g/ml$ concentration for 1.5 hours significantly increased than that of control in cell attachment, and cell process is also evident compared with that of control in cell morphology, and the cellular activity and DNA synthesis rate of cells treated minocycline and $TGF-{\beta}1$ significantly increased than that of control values, but were below to values of the $TGF-{\beta}1$ only treated group in MIT assay and $^3H-thymidine\;assay$, and the total protein synthesis of minocycline and $TGF-{\beta}1$ treated group also significantly increased than that of control values, but the percent collagen synthesis of tested group significantly decreased to compared with control. On the above the findings, the tested group of minocycline and $TGF-{\beta}1$ did not increase the effect on the cell activity than $TGF-{\beta}1$ only tested group and the tested group of minocycline inhibited cell activity. This results indicate that minocycline was effective on cell attachment in early stage, but it is harmful to cell activity, that inhibitory effect of minocycline was compensated with stimulatory effect of $TGF-{\beta}1$.

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