• Title/Summary/Keyword: tissues.

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The effect of tumor necrosis factor (TNF)-α to induce matrix metalloproteinase (MMPs) from the human dental pulp, gingival, and periodontal ligament cells (사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구)

  • Rhim, Eun-Mi;Park, Sang-Hyuk;Kim, Duck-Su;Kim, Sun-Young;Choi, Kyoung-Kyu;Choi, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.36 no.1
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    • pp.26-36
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    • 2011
  • Objectives: In the present study, three kinds of tissues cells (pulp, gingiva, and periodontal ligament) were investigated if those cells express MMP and TIMP when they were stimulated with neuropeptides (substance P, CGRP) or proinflammatory cytokine, TNF-$\alpha$. Materials and Methods: The cells cultured from human dental pulp (PF), gingiva (GF) and periodontal ligament were (PDLF) stimulated with Mock, SP, TNF-$\alpha$, and CGRP for 24 hrs and 48 hrs. for an RNase protection assay and Enzyme Linked Immunosorbent Assay. Cells (PF, GF and PDLF) seeded in 100 mm culture dish were stimulated with SP ($10^{-5}$, $10^{-8}\;M$) or only with medium (Mock stimulation) for 4hrs and for 24 hrs for RNase Protection Assay, and they were stimulated with CGRP ($10^{-5}\;M$) and TNF-$\alpha$(2 ng/mL) for 24 hrs and with various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for Rnase Protection Assay with a human MMP-1 probe set including MMP 1, 2, 8, 7, 8, 9, 12, and TIMP 2, 3. In addition, cells (PF, GF and PDLF) were stimulated with Mock and various concentraion of TNF-$\alpha$(2, 10, and 100 ng/mL) for 24 hrs and with TNF-$\alpha$(10 ng/mL) for 48 hrs, and the supernatents from the cells were collected for Enzyme Linked Immunosorbent Assay (ELISA) for MMP-1 and MMP-13. Results: The expression of MMPs in PF, GF, PDLF after stimulation with SP and CGRP were not changed compared with Mock stimulation for 4 hrs and 24 hrs. The expression of MMP-1, -12, -13 24 hrs after stimulation with TNF-$\alpha$ were upregulated, however the expression of TIMP-3 in PF, GF, PDLF after stimulation with TNF-$\alpha$ were downregulated. TNF-$\alpha$(2 ng/mL, 10 ng/mL, 100 ng/mL) increased MMP-1 and MMP-12 expression in PF dose dependently for 24 hrs. Conclusions: TNF-$\alpha$ in the area of inflammation may play an important role in regulating the remodeling of dentin, cementum, and alveolar bone.

Effects of Ca Supplementation on Mineral Metabolism during Pregnancy and Lactation of Calcium-Deficient Young Adult Rats (칼슘섭취 부족 흰쥐에서 임신, 수유기 동안 칼슘보충 수준이 무기질 대사에 미치는 영향)

  • 박미나;김은애;이연숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.6
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    • pp.958-964
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    • 2004
  • This study examined the effects of maternal calcium intake levels on mineral metabolism during pregnancy and lactation. Five week-old female Sprague-Dawley rats were fed normal Ca (NCa, 0.5%) and low Ca (LCa, 0.15%) diets for 5 weeks during preconception. The low Ca intake group was divided into three groups and fed low Ca (0.15%), normal Ca (0.5%) and high Ca (1.5%) diets for 6 weeks during pregnancy and lactation. All of the rats were mated with normal male rats. The control group was consistently fed a normal Ca diet for 11 weeks. Pups were adjusted to 10 per dam. After weaning, dams and their weaned rats were sacrificed. We measured total protein, albumin, alkaline phosphatase and mineral content in serum and weight, length, breaking force, ash and mineral content in bone and kidney. There was no significant difference in weight gain and food intake among the groups. Serum total protein and albumin were in the normal range but slightly decreased with a low Ca diet during preconception. Calcium supplementation above normal level increased bone weight, Ca content and breaking force in femur of maternal and their weaned rats. However, high maternal Ca intake significantly decreased Mg and Fe contents in maternal serum and bone and Mg and Zn contents in serum of weaned rats. Also, it induced Ca, P and Mg deposit in maternal kidney. Our experimental results suggest that Ca deficiency during preconception may deteriorate bone status of mother and their offsprings. Moreover, high Ca supplementation during pregnancy and lactation may lead to disturb mineral metabolism in bone and other tissues.

Effects of Chronic and Acute Stress on Clusterin Secretion of the Rat Submandibular Gland (급만성 스트레스가 백서 악하선의 Clusterin 분비에 미치는 영향)

  • Jin, Sang-Bae;Chun, Yang-Hyun;Hong, Jung-Pyo
    • Journal of Oral Medicine and Pain
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    • v.31 no.1
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    • pp.79-89
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    • 2006
  • The aim of this study is to know how the rat submandibular gland changes under various emotional stress condition, using molecular biological methods. Restraint and chronic unpredictable mild stress (CUMS) experiment is conducted on fifty one 7-week old Sprague-Dawley rats (restraint stress experiment: 21, CUMS: 30). The rats were sacrificed, the submandibular glands were excised immediately at certain time, and examined by the use of immunohistochemistry and western blotting. In CUMS experiment, sucrose preference test, water intake change, weight change were implemented at 1 week interval for the experimental period The results are as follows: 1. The number of clusterin-secreting cells of restraint stress group compared to control group showed significantly decreasing tendency in all experimental groups except for the 1st hour group (p<0.001 in the 9th, 24th, 72nd, 120th, and 168th hour group). 2. The number of clusterin-secreting cells of CUMS group compared to control group showed significantly increasing tendency in the 2nd week group (p<0.01), and significantly decreasing tendency in the 4th and 5th week group (p<0.001). 3. Sucrose preference test in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01). 4. Weight change in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01), but water intake change didn't show significant difference compared to control group. 5. In western blot analysis, clusterin expression was decreased on a gradual basis in due time compared to the control group in the restraint stress group. As for CUMS group (chronic unpredictable mild stress group), it was increased till the 2nd week and decreased till the 5th week after that, which is similar to immunohistochemical analysis result and the decreasing tendency of sucrose preference and weigh changes. Through the test, it was proved that expression of clusterin in saliva glands decreases after receiving either acute or chronic stress, indicating relation with depression caused by chronic stress. Unlike other data, however, apoptotic tendency was hardly found in tissues. Diverse possibilities could be suggested on that: first, the stress was not enough to expedite apoptosis; second, apoptosis-related protein was already being secreted though not detected with microscope; third, clusterin, a major secretion molecule of saliva, decreased with saliva's malfunction due to stress. In the respect, it will be necessary to examine proteins expressed in case of cell death or other heat-shock proteins at the same time, in order to see whether any cellular change or death is caused by decreasing clusterin under high stress, and whether the original state is restored as time goes by under mild stress, through longer-term tests using even higher acute stress.

Activation and Abnormalities of Cell Cycle Regulating Factor in Head and Neck Squamous Cell Carcinoma Cell Lines: Abnormal Expression of CDKN2 Gene in Laryngeal Squamous Cell Carcinoma (두경부 편평상피세포암 세포주에서 세포주기조절인자의 활성 및 이상 : 후두편평상피세포암에서 종양억제유전자 CDKN2 유전자의 발현이상)

  • Song, Si-Youn;Han, Tae-Hee;Bai, Chang-Hoon;Kim, Yong-Dae;Song, Kei-Won
    • Journal of Yeungnam Medical Science
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    • v.22 no.2
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    • pp.166-182
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    • 2005
  • Background: Cyclin-dependent kinase (CDK) inhibitors are family of molecules that regulate the cell cycle. The CDKN2, a CDK4 inhibitor, also called p16, has been implicated in human tumorigenesis. The CDKN2 inhibits the cyclin/CDK complexes which regulate the transition from G1 to S phase of cell cycle. There is a previous report that homozygous deletion of CDKN2 region on chromosome 9p21 was detected frequently in astrocytoma, glioma and osteosarcoma, less frequently in lung cancer, leukemia and ovarian cancer, but not detected in colon cancer and neuroblastoma. However, little is known about the relationship between CDKN2 and laryngeal cancer. Therefore this study was initiated to investigate the role of CDKN2 in human laryngeal squamous cell carcinoma development.1) Materials and methods: We used 5 human laryngeal carcinoma cell lines whether they have deletions or losses of CDKN2 gene expression by DNA-PCR or RT-PCR, respectively. We examined 8 fresh frozen human laryngeal cancer tissues to detect the loss of heterozygosity (LOH) of CDKN2. PCR was performed by using microsatellite markers of short arm of human chromosome 9 (D9S126, D9S144, D9S156, D9S161, D9S162, D9S166, D9S171, D9S200 and D9SIFNA). For informative cases, allelic loss was scored if the signal of one allele was significantly decreased in tumor DNA when compared to the same allele in normal DNA. Results: The CDKN2 DNA deletion was observed in 3 cell lines. The CDKN2 mRNA expression was observed in only one cell line, which was very weak. LOH was detected in 7 cases (87.5%). Conclusion: These results suggest that CDKN2 plays a role in the carcinogenesis of human laryngeal squamous cell carcinoma.

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Comparison and Evaluation of radiotherapy plans by multi leaf collimator types of Linear accelerator (선형가속기의 다엽콜리메이터 형태에 따른 치료계획 비교 평가)

  • Lim, Ji Hye;Chang, Nam Joon;Seok, Jin Yong;Jung, Yun Ju;Won, Hui Su;Jung, Hae Youn;Choi, Byeong Don
    • The Journal of Korean Society for Radiation Therapy
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    • v.30 no.1_2
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    • pp.129-138
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    • 2018
  • Purpose : An aim of this study was to compare the effect of multi leaf collimator(MLC) types for high dimension radiotherapy in treatment sites used clinically. Material and Method : 70 patients with lung cancer, spine cancer, prostate cancer, whole pelvis, head and neck, breast cancer were included in this study. High definition(HD) MLC of TrueBeam STx (Varian Medical system, Palo Alto, CA) and millenium(M) MLC of VitalBeam (Varian Medical system, Palo Alto, CA) were used. Radiotherapy plans were performed for each patient under same treatment goals with Eclipse (Version 13.7, Varian Palo Alto USA, CA). To compare the indicators of the radiotherapy plans, planning target volume(PTV) coverage, conformity index(CI), homogeneity index(HI), and clinical indicators for each treatment sites in normal tissues were evaluated. To evaluate low dose distribution, $V_{30%}$ values were compared according to MLC types. Additionally, length and volume of targets for each treatment sites were investigated. Result : In stereotatictic body radiotherapy(SBRT) plan for lung, the average value of PTV coverage was reduced by 0.52 % with HD MLC. With SBRT plan using HD MLC for spine, the average value of PTV coverage decreased by 0.63 % and maximum dose decreased by 1.13 %. In the test of CI and HI, the values in SBRT plan with HD MLC for spine were 1.144, 1.079 and the values using M MLC were 1.160, 1.092 in SBRT plan for lung, The dose evaluation of critical organ was reduced by 1.48 % in the ipsilateral lung mean dose with HD MLC. In prostate cancer volumetric modulated arc therapy(VMAT) with HD MLC, the mean dose and the $V_{30}$ of bladder and the mean dose and the $V_{25}$ of rectum were reduced by 0.53 %, 1.42 %, 0.97 %, and 0.69 %, respectively (p<0.05). The average value of heart mean dose was reduced by 0.83 % in breast cancer VMAT with M MLC. Other assessment indices for treatment sites showed no significant difference between treatment plans with two types of MLC. Conclusion : Using HD MLC had a positive impact on the PTV coverage and normal tissue sparing in usually short or small targets such as lung and spine SBRT and prostate VMAT. But, there was no significant difference in targets with long and large such as lung, head and neck, and whole pelvis for VMAT.

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Evaluation of the Jaw-Tracking Technique for Volume-Modulated Radiation Therapy in Brain Cancer and Head and Neck Cancer (뇌암 및 두경부암 체적변조방사선치료시 Jaw-Tracking 기법의 선량학적 유용성 평가)

  • Kim, Hee Sung;Moon, Jae Hee;Kim, Koon Joo;Seo, Jung Min;Lee, Joung Jin;Choi, Jae Hoon;Kim, Sung Ki;Jang, In-Gi
    • The Journal of Korean Society for Radiation Therapy
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    • v.30 no.1_2
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    • pp.177-183
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    • 2018
  • Purpose : Volumetric Modulated Arc Therapy(VMAT) has the advantage of uniformly and precisely irradiating the tumor to the shape of the tumor while reducing the risk of radiation damage to normal tissues. such as brain cancer, head and neck cancer and prostate cancer, It is being used for treatment. The purpose of this study is to evaluate the usefulness of the Jaw-Tracking technique(JTT) in VMAT for brain and head and neck cancer. Materials and Methods : We selected eight patients with brain and head and neck cancer(4 Brain, 4 head and neck) who were treated with the VMAT treatment technique. Contouring information of the patient's tumor and normal organ was fused to the Rando phantom using the deformable registration of Velocity(Varian, USA). A treatment plan was developed using the Varian Eclipse(ver 15.5, Varian, USA) with the same patient actual beam parameters except for the use of jaw-tracking. As the evaluation index, the maximum dose and mean dose of target and OAR were compared and a portal dosimetry was performed for the treatment plan verification. Results : When using JTT, the relative dose of OAR decreased by 5.24 % and the maximum dose by 7.05 %, respectively, compared with the Static-Jaw technique(SJT). In the various OARs, the mean dose and maximum dose reduction ranges ranged from 0.01 to 3.16 Gy and from 0.12 to 6.27 Gy, respectively. In the case of the target, the maximum dose of GTV, CTV, PTV decreased by 0.17 %, 0.43 %, and 0.37 % in JTT, and the mean dose decreased by 0.24 %, 0.47 % and 0.47 %, respectively. Gamma analysis The JTT and SJT passing rates were $98{\pm}1.73%$ and $97{\pm}1.83%$ on the basis of 3 % / 3 mm, respectively. Comparing the doses of all OARs applied to the experiment, it was found that the use of JTT resulted in a significant decrease in dose due to additional jaw shielding besides MLC than SJT. Conclusion : In radiation therapy using VMAT treatment plan, we can apply JTT in the case of adjacent tumor and normal organs such as brain cancer and head and neck cancer, and in radiotherapy required large field and high energy caused increase leakage dose through MLC. It is considered that the target dose of PTV can be increased by lowering the dose of normal tissue surrounding the tumor.

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Expression of Decidual Natural Killer (NK) Cells in Recurrent Abortion Patients with Increased Peripheral NK Cells (말초혈액 자연살해세포가 증가된 반복유산 환자의 탈락막 자연살해세포의 발현)

  • Lee, Hyun-Joo;Kim, Sun-Haeng;Yang, Kwang-Moon;Park, Dong-Wook;Park, Chan-Woo;Hong, Sung-Ran;Cho, Dong-Hee;Song, In-Ok;Koong, Mi-Kyoung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.199-207
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    • 2009
  • Objectives: The purpose of this study was to compare the decidual NK cell populations between increased pre-conceptional peripheral NK cell population and normal pre-conceptional peripheral NK cell population in women with a history of recurrent abortion. Methods: Fourteen women with history of recurrent abortion and elevated pre-conceptional peripheral NK cell, above 15% of peripheral lymphocyte population were included in this study. As a control, twelve women with history of recurrent abortion and their peripheral NK cell percentage showed below 15% were included. Distribution of $CD56^+$ and $CD16^+$ NK cells in paraffin embedded decidual tissues including implantation sites were examined by immunohistochemical staining using anti-CD56, 16 monoclonal antibodies. After immuohistochemical staining, the numbers of decidual NK cells were counted and compared these results between study and control groups. Results: There was significant difference in decidual $CD56^+$ NK cell count ($170.1{\pm}132.1$ vs. $68.3{\pm}66.1$, p=0.02) between increased peripheral $CD56^+$ NK cell group and control group. But, there showed no statistically significant correlation between decidual $CD56^+$ NK cell count and peripheral $CD56^+$ NK cell percentage (r=0.229, p=0.261). Also there was no statistically difference decidual $CD16^+$ NK cell count between study and control group ($25.70{\pm}11.72$ vs. $31.17{\pm}22.67$), and no correlation between decidual $CD16^+$ NK cell and peripheral $CD16^+$ NK cell percentage (r=-1.40, p=0.535). Conclusions: This study shows that decidual $CD56^+$ NK cell are significantly increased in decidua of women exhibiting a history of recurrent abortion with increased $CD56^+$ peripheral NK cell. This study suggests that the percentage of peripheral NK cell reflect the expression of decidual NK cell. Consequently, pre-conceptional peripheral blood NK cell population can be the useful marker for detecting the risk of subsequent miscarriage.

The inhibitory Effects of Coenzyme Q10 on Melanogenesis of cultured Human Melanocytes and in vivo Guinea Pig Model (Coenzyme Q10의 멜라닌 생성억제효과)

  • 황재성;박원만;안수미;강병영;이병곤;심영철
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.149-162
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    • 2000
  • Coenzyme Q10 is found in all tissues including skin and it is the well-known coenzyme for mitochondrial enzymes. The electron and proton transfer functions of the quinone ring are of fundamental importance for the oxidative phosphorylation pathway to generate energy in the cells. Coenzyme Q10 has been studied as a potent antioxidant molecule in the skin. It is involved in the skin's response to UVR irradiation. The concentration of this antioxidant in UVR exposed skin is higher than in non-exposed skin. However, recent studies have also shown that coenzyme Q10 is one of the first antioxidants to be depleted when skin is UVR-irradiated. This indicates that coenzyme Q10 is primarily involved in defense mechanisms of the skin. Therefore, we questioned whether coenzyme Q10 shows reulatory effect of melanogenesis. Here we report that coenzyme Q10 inhibits melanin neosynthesis of normal human melanocytes grown in culture, and lightens UVB-induced hyperpigmentation of the guinea pig skin in vivo. We treated human melanocytes with 0.05mM to 0.5mM of coenzyme Q10 for a total of two days. This inhibited melanin neosynthesis of cultured human melanocytes dose-dependently. The inhibitory effect of coenzyme Q10 was as effective as kojic acid or vitamin C on cultured human melanocytes. CoQ10 didn't have direct inhibitory effect on tyrosinase activity in in vitro tyrosine hydroxylase activity To further clarify the effect of coenzyme Q10 on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brownish guinea pigs. The UVB intensity was 500mJ/$\textrm{cm}^2$ and the total energy dose was 1,500 mJ/$\textrm{cm}^2$. The animals were exposed to UVB radiation one times a week for three consecutive weeks. Coenzyme Q10, kojic acid, Arbutin, vitamin C(1% in vehicle) or vehicle alone as a control were then topically applied daily to the hyperpigmented areas twelve times per week far four successive weeks. The lightening effect was evaluated by visual scoring, chromameter and immunohistochemistry. Coenzyme Q10 had lightening effect on the UVB-induced hyperpigmentation without any other side effects, whereas another compounds showed weak lightening efficacies. Therefore, these results suggest that coenzyme Q10 may be useful for solving physiological hyperpigmenting problems for cosmetic purposes.

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Isolation and Characterization of Elastase Inhibitor from Areca catechu (빈랑으로부터 Elastase 저해물질의 분리 및 특성조사)

  • 조중제;이건국;조병기;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.26 no.1
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    • pp.163-186
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    • 2000
  • We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

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Polarization-sensitive Optical Coherence Tomography Imaging of Pleural Reaction Caused by Talc in an ex vivo Rabbit Model (생체 외 토끼 모델에서의 탈크에 의해 유발되는 흉막 반응의 편광 민감 광 결맞음 단층촬영 이미징)

  • Park, Jung-Eun;Xin, Zhou;Oak, Chulho;Kim, Sungwon;Lee, Haeyoung;Park, Eun-Kee;Jung, Minjung;Kwon, Daa Young;Tang, Shuo;Ahn, Yeh-Chan
    • Korean Journal of Optics and Photonics
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    • v.31 no.1
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    • pp.1-6
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    • 2020
  • The chest wall, an organ directly affected by environmental particles through respiration, consists of ribs, a pleural layer and intercostal muscles. To diagnose early and treat disease in this body part, it is important to visualize the details of the chest wall, but the structure of the pleural layer cannot be seen by chest computed tomography or ultrasound. On the other hand, optical coherence tomography (OCT), with a high spatial resolution, is suited to observe pleural-layer response to talc, one of the fine materials. However, intensity-based OCT is weak in providing information to distinguish the detailed structure of the chest wall, and cannot distinguish the reaction of the pleural layer from the change in the muscle by the talc. Polarization-sensitive OCT (PS-OCT) takes advantage of the fact that specific tissues like muscle, which have optical birefringence, change the backscattered light's polarization state. Moreover, the birefringence of muscle associated with the arrangement of myofilaments indicates the muscle's condition, by measuring retardation change. The PS-OCT image is interpreted from three major perspectives for talc-exposure chest-wall imaging: a thickened pleural layer, a separation between pleural layer and muscle, and a phase-retardation measurement around lesions. In this paper, a rabbit chest wall after talc pleurodesis is investigated by PS-OCT. The PS-OCT images visualize the pleural layer and muscle, respectively, and this system shows different birefringence of normal and damaged lesions. Also, an analyisis based on phase-retardation slope supports results from the PS-OCT image and histology.