• Title/Summary/Keyword: in vitro experiments

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Comparison of Growth Characteristics, Forage Productivity and Quality between Italian Ryegrass and Synthetics Derived from Interspecific and Intergeneric Hybrids and Ecotypes (이탈리안 라이그라스와 종.속간 잡종 및 생태형 유래 합성종간의 생육특성, 수량성 및 사료가치 비교)

  • Choi, G.J.;Lim, Y.W.;Lim, Y.C.;Sung, B.R.;Kim, M.J.;Kim, K.Y.;Park, G.J.;Kim, S.R.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.4
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    • pp.265-272
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    • 2002
  • Forage breeding laboratory of National Livestock Research Institute, R.D.A. has made interspecific hybrids of Lolium multiflorum $\times$ L. pratensis and intergeneric hybrids of Lolium $\times$ Festuca since 1984, and has collected ecotypes of Italian ryegrass since 1991. Growth characteristics of these hybrids and ecotypes were researched, and then these clone lines were named. Among these clone lines, the several clones that have polen fertility, high cold-tolerance, and similar heading time were used for making synthetics, Naehan 6, 7, 8, 9, with polycrossing method in 1997. Field experiments were carried out to compare the mophological and agronomical characteristics and forage productivity and quality of the synthetics with those of Italian ryegrass varieties, Barmultra and Hwasan 101. in Suwon and Yonchun from 1999 to 2000. Heading time of the synthetics were 22th to 24th May that belong to late-mature types to be similar to that of Barmultra and Hwasan 101 in Suwon. The synthetics were 101 to 106 c3n in plant length, medium or thick in thickness of stem, dark peen in leaf color, broad and long in flag leaf, strong in lodging resistance, and excellent in regrowth. Winter survivals of the synthetics were no different from that of Barmultra or Hwasan 101 in Suwon, but better than that of Barmultra or Hwasan 101 in Yonchun where was -10 to -12$^{\circ}C$ of minimum average air temperature in January or February. Dry matter(DM) yields of the synthetics were similar to DM 8,238kg per ha of Barmultra in Suwon, but in Yonchun, were more 7 to 13% than DM 7,291kg per ha of Barmultra. Forage qualities, IVDMD, ADF, NDF and TDN of the synthetics were lower than those of Hwasan 101, but higher than those of Barmultra.

The Effect of Safflower Seed Extract on the Bone Formation of Calvarial Bone Model in Sprague Dawley rat (백서 두개골 결손부에서 홍화씨 추출물의 골조직 재생 유도 효과)

  • Kim, Sung-Tae;Jhon, Gil-Ja;Lim, So-Hyoung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.835-852
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    • 2000
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and repair of function. For more than a decade there have been many efforts to develop materials and methods of treatment to promote periodontal wound healing. Recently many efforts are concentrated on the regeneration potential of material used in oriental medicine. In some in vitro and in vivo experiments, there have been many evidences that these materials have an effect on bone regeneration. The purpose of this study was to evaluate histologically and radiologically in Sprague-Dawley rats the effects of safflower seed extracts on the regeneration of the calvarial defects surgically produced. So in this study, the critical size defects were surgically produced in the calvarial bone of 30 Sprague-Dawley rats using the 8mm trephine bur. The safflower seed extract was applied into the defect of each rat in experimental group, whereas nothing was applied into the defect of each rat in control group. Rats were sacrificed at 2, 4, 8 weeks following operation and histomorphometric and radiodensitometric analysis were performed. 1. The newly formed bone length was $102.91{\pm}22.05$, $178.29{\pm}24.40$ at 2 week in the each control, experimental group, $130.95{\pm}39.24$, $242.62{\pm}50.33$ at 4 week and $181.53{\pm}76.35$, $240.36{\pm}22.00$ at 8 week($unit,{\mu}m$). In the 2, 4 week, there were statistically significant difference between control and experimental group(P<0.05). 2. The newly formed bone area was $2962.06{\pm}1284.48$, $10648.35{\pm}1284.48$ at 2 week, $5103.25{\pm}1375.88$, $9706.78{\pm}1481.81$ at 4 week, $8046.02{\pm}818.99$, $12057.06{\pm}740.47$ at 8 week($unit,{\mu}m^2$). In every week, there were statistically significant difference between control and experimental group(P<0.05). 3. The radiopacity was $14.26{\pm}.33$, $25.47{\pm}4.33$ at 2 week, $20.06{\pm}9.07$, $26.61{\pm}2.78$ at 4 week, $22.99{\pm}3.76$, $27.29{\pm}1.54$ at 8 week(unit, %). In the 2 week, there was statistically significant difference between control and experimental group(P<0.05). In conclusion, the results of the present study suggest that safflower seed extract initially has an effect on the newly formed bone area, length and radiopacity when it is applied to the calvarial defect of Sprague - Dawley rat. Then. the material has an effect on newly formed bone area and length.

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Study on the Safety of Firefly Luciferase in Human as a Transient Reporter Gene of Oncolytic Virotherapy (항암 바이러스 치료제의 보고유전자로써 반딧불이 루시퍼레이즈의 인체 내 안전성에 대한 연구)

  • Hong, Young Mi;Yoon, Woong Hee;Lee, You Ra;Kim, Soo Ji;Ngabire, Daniel;Narayanasamy, Badrinath;Ornella, Mefotse Saha Cyrelle;Kim, Myunghee;Cho, Euna;Lee, Bora;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.31 no.11
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    • pp.1028-1036
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    • 2021
  • Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

THE EFFECTS OF CITRIC ACID TREATED ROOT SURFACES ON THE ATTACHMENT AND PROLIFERATION OF PERIDONTAL LIGAMENT CELLS (치근면 구연산 도포가 치주인대세포의 부착과 전개에 미치는 영향)

  • Lee, Sang-Gu;Suh, Jo-Young;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.77-96
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    • 1993
  • This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

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The effect of admixture of vitamin $D_3$ and dexamethasone on the activity of osteoblastic cells (Vitamin $D_3$와 Dexamethasone의 복합 투여가 골모세포에 미치는 영향에 관한 연구)

  • Lim, Na-Won;Park, Young Joo;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.29 no.3 s.74
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    • pp.383-397
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    • 1999
  • Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.

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Effects of Molasses Addition on the Feeding Value of Socheongryongtang(Herbal Medicine) Meal Silage (당밀 첨가가 소청룡탕박 사일리지의 품질에 미치는 영향)

  • Moon, Kye Bong;Cho, Sung Kyung;Park, Duk Sub;Kim, Sung Bok;Lee, Bong Duk;Jo, Cheorun;Lim, Jae Sam;Lee, Soo Kee
    • Journal of Animal Science and Technology
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    • v.50 no.3
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    • pp.381-390
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    • 2008
  • Two experiments were conducted to investigate effects of molasses addition to silage materials in ensiling Socheongryongtang meal on the nutritive quality of silage, palatability and various ruminal parameters in Korean native goats. In Experiment 1, Socheongryongtang meal silage was produced by the addition of 0, 0.5, or 1.0% molasses and stored for 40 days at room temperature. There were three replicates per treatment. Lactic acid contents of Socheongryongtang meal silage containing molasses(0.5 and 1.0%) were significantly(p<0.05) higher than that of the control(0%). However, the pH and butyric acid contents of Socheongryongtang meal silage containing molasses(0.5 and 1.0%) were lower than those of the control(0%). In addition, molasses(0.5 and 1.0%) increased the number of lactobacillus, but decreased the number of fungi in Socheongryongtang meal silage. In vitro dry matter disappearance tended to increase by molasses addition. In Experiment 2, three 1.5-yr-old Korean native female goats were employed in cross-over design to measure the palatability and various ruminal parameters of Socheongryongtang meal silage. Feed intake of Socheongryongtang meal silage containing molasses(0.5 and 1.0%) for 30 min significantly higher than that of control(0%). Silmilar trend was found when the feed intake was measured for 6 hr. In various ruminal parameters, molasses addition to silage materials increased propionic acid(P) and total volatile fatty acid contents, but decreased pH, acetic acid (A), and butyric acid. The A/P ratio was the lowest in 1.0% molasses treatment group. It is concluded that molasses addition in ensiling Socheongryongtang meal could enhance its nutritional quality and lengthen storage period. Also the palatability of silage was improved by the addition of molasses.

A Study on the Effect of Sample Storage Condition on the RIA Results of Plasma renin activity Test (Plasma renin activity 검사의 검체 보관 방법이 방사면역 측정법 결과에 미치는 영향에 대한 고찰)

  • Choe, Jin-joo;Back, Song-ran;Yoo, Seon-hee;Lee, Sun-ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.25 no.1
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    • pp.29-33
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    • 2021
  • Purpose Plasma renin activity (PRA) test is important for the diagnosis of primary aldosteronism. PRA is an easily deformed substance in vitro and affected by temperature changes. Laboratory of ASAN medical center has consistently found that there was a difference between the initial and re-experimental results. We compared and analyzed the differences in PRA test results according to the sample storage status. Materials and Methods The measurement of PRA was performed by using the radioimmunoassay. From August to September 2020, 43 PRA re-test samples were tested with different sample storage condition. The first group was re-examined by freezing the plasma-separated samples at -18℃, and the second group was re-examined with refrigerated EDTA sample. Also, additional tests were conducted on 13 PRA samples to verify the effect on thawing temperature differences in plasma-separated samples. The same samples were divided into two parts and stored frozen at -18℃, respectively, and thawing samples in room temperature and those in refrigerator were were conducted. Each result was compared and analyzed based on the initial experimental results. Results The results of re-examination after frozen storing plasma separation samples showed a lower correlation than the results of re-examination with EDTA plasma samples in refrigerator. When calculating the percentage based on the initial test results, the average percentage of each was 404.9% and 133.8%. The correlation coefficient was also R=0.8501 and R=0.9966, respectively, showing a higher correlation between plasma in the refrigerated sample EDTA tube. In comparison experiments with differences in thawing temperature, average percentage of the results of initial test and room temperature thawing was 94.3% and the average percentage of the results of refrigerated thawing was 88.0%. After again freezing the sample, the average percentage of the second room temperature thawing result is 107.5%, and the second refrigerated thawing group is 112.7%. Both groups showed an increase from first thawing. Conclusion A comparative analysis of retesting according to differences in sample storage methods in PRA tests showed a higher correlation between the results of retesting of the refrigerated EDTA plasma. And repeated freezing and melting of plasma separation samples, regardless of temperature during defrosting, has been shown to affect results. Therefore, retest of PRA should re-collect plasma from original EDTA plasma to increase reproducibility.

Whitening activity of Ficus carica L. fruits extract through inhibition of tyrosinase and MITF expression (무화과(Ficus carica L.) 열매 추출물의 tyrosinase 및 MITF 발현 억제를 통한 미백 활성)

  • Min Ji Kim;Si Eun Park;Geun soo Lee;Jin Hwa Kim;Sunwoo Kwon;Hyung Seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.204-212
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    • 2023
  • Whitening is inhibitory activity of the melanin synthesis of melanocytes. Recently, whitening materials have been developed on natural materials because of its side effects on skin. Figs (Ficus Carica L.) is a fruit belonging to the Moraceae family and whitening activity was reported in focusing on the fig's stem and leaf components, but whitening activity of the figs fruit was not known. Thus, in this study, we tried to observe its anti-melanogenesis as well as antioxidant and anti-inflammation. The radical scavenging activity of figs fruits extract (FFE) was observed as the level of 34.52±1.98%/60.71±1.26% compared to the control in the its maximum concentration in the DPPH/ABTS assay. Cytotoxicity of FFE was observed at 10% concentration by CCK8 assay, so the maximum concentration was set at 5% and applied to all experiments. FFE concentration dependently decreased NO production associated with inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-α gene expression, these strongly suggesting anti-inflammatory activity. In melanin contents assay, FFE significantly down-regulated melanin production in α-MSH-stimulated B16F10 cell as well as tyrosinase inhibition in vitro. In addition, FFE decreased the Microphthalmia-associated transcription factor (MITF) mRNA expression about 94.34% compared to the α-MSH treatment group in RT-PCR. Finally, FFE significantly reduced the MITF, cAMP response element-binding protein and tyrosinase protein expression in the α-MSH stimulated B16F10 cell. Through these results, we found that FFE can not only directly inhibit tyrosinase enzyme activity but also suppress melanogenesis through regulation of MITF gene expression in α-MSH signal transduction.

Effect of Glucose Exposure on the Development of the Mouse Preimplantation Embryo In Vitro (착상전 생쥐배아의 Glucose에 대한 노출이 체외 배발생에 미치는 영향)

  • Kim, Seon-Ui;Eom, Sang-Jun;Yun, San-Hyeon;Im, Jin-Ho;Jeong, Gil-Saeng
    • Korean Journal of Animal Reproduction
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    • v.19 no.3
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    • pp.227-234
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    • 1995
  • This study was carried out to investigate the effects according to the time course of glucose exposure on the development of one-cell embryos beyond morula in CR$_laa$ medium. One-cell zygotes from B6CBA F$_1$ mice were recovered at 24 ~ 25h after hCG and cumulus cells were removed with 0.1% hyaluronidase. The embryos were pooled and subsequently divided into each groups and cultured in CR$_laa$ at 37$^{\circ}C$ in 5% CO$_2$ in aIr. The embryos were either, placed in CR$_laa$ containing various concentration (5.5, 16.5, 27.5 and 38.5 mM) of glucose for 1 min. and subsequently returned to the fresh culture medium (without glucose), or were transferred to the same media containing glucose at 72 h post hCG. The results obtained in these experiments were summarized as follows: 1. The development rates of zygotes, recovered from the oviducts in M2 and cultured in CR$_laa$ with 3mg/Im FAF-BSA, to expanded blastocysts (25.7%) and hatching bIastocysts (17.6%) were significantly higher than those of zygotes recovered in TL Hepes (0% and 0%, respectively). 2. The development rates of one-cell embryos exposed to 27.5 mM glucose at 72 h post hCG for 1 min, were 68.8% (CR$_1$+BSA) a and 77.1% (CR$_1$+FBS) of expanded blastocyst stage, but there were no significant differences between the embryos exposed for 1 min. or transferred at 72 h. 3. Regardless of glucose concentration (5.5, 16.5, 27.5 & 38.5mM), 45.7~61.5% of embryos developed to the blastocyst stage. There were no significant differences between any of the treatments on the devel-opment of one-cell embryos. Therefore, the detrimental effect of highly concentration was not appeared.

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THE EFFECT OF LACTIC ACID CONCENTRATION AND pH OF LACTIC ACID BUFFER SOLUTIONS ON ENAMEL REMINERALIZATION (완충 용액의 유산 농도와 pH가 법랑질의 재광화에 미치는 영향)

  • Kwon, Jung-Won;Suh, Duk-Gyu;Song, Yun-Jung;Lee, Yun;Lee, Chan-Young
    • Restorative Dentistry and Endodontics
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    • v.33 no.6
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    • pp.507-517
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    • 2008
  • There are considerable in vitro and in vivo evidences for remineralization and demineralization occurring simultaneously in incipient enamel caries. In order to "heal" the incipient dental caries, many experiments have been carried out to determine the optimal conditions for remineralization. It was shown that remineralization is affected by different pH, lactic acid concentrations, chemical composition of the enamel, fluoride concentrations, etc. Eighty specimens from sound permanent teeth without demineralization or cracks, 0.15 mm in thickness, were immersed in lactic acid buffered demineralization solutions for 3 days. Dental caries with a surface zone and subsurface lesion were artificially produced. Groups of 10 specimens were immersed for 10 or 12 days in lactic acid buffered remineralization solutions consisting of pH 4.3 or pH 6.0, and 100, 50, 25, or 10 mM lactic acid. After demineralization and remineralization, images were taken by polarizing microscopy (${\times}100$) and micro-computed tomography. The results were obtained by observing images of the specimens and the density of the caries lesions was determined. 1.As the lactic acid concentration of the remineralization solutions with pH 4.3 was higher, the surface zone of the carious enamel increased and an isotropic zone of the subsurface lesion was found. However, the total decalcification depth increased at the same time. 2.In the remineralization solutions with pH 6.0, only the surface zone increased slightly but there was no significant change in the total decalcification depth and subsurface zone. In the lactic acid buffer solutions with the lower pH and higher lactic acid concentration, there were dynamic changes at the deep area of the dental carious lesion.