• Journal of Periodontal and Implant Science
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• 제36권2호
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• pp.335-344
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• 2006

Osteoblast or bone marrow stromal cell-derived RANKL is the major effector molecule essential for osteoclastogenesis. Previous studies have shown that tetracyclines have beneficial therapeutic effects in the prevention and treatment of inflammatory bone disease including periodontal disease. Periodontal ligament cells are thought not only to play an important role in the progression of periodontal disease, but to play an important role in alveolar bone remodeling. Previous studies indicated that receptor activation of nuclear factor $\kappa\;B$ ligand (RANKL) and osteoprotegerin (OPG) are expressed in periodontal ligament cells by pro-inflammatory cytokine, such as $IL-1{\beta}$ and $TNF-{\alpha}$. This study was designed to investigate the inhibitory effect of doxycycline on RANKL and OPG mRNA in rat periodontal ligament cells induced by $IL-1{\beta}$ (1 ng/ml). The results are as follows; 1. MTT assay showed that doxycycline at the concentration of $1-50\;{\mu}g/m{\ell}$ didn't result in statistically significant cell death at day 1 and 3. 2. RANKL mRNA expression was increased to 2.6 folds by $IL-1{\beta}$. When cells were treated with doxycycline ($50{\mu}g/m{\ell}$), $IL-1{\beta}$ -induced mRANKL expression was reduced by 33%. In contrast to RANKL, OPG mRNA expression was not inhibited by pre-treatment with doxycycline. These results suggest that doxycycline decrease the expression of mRANKL resulting in regulation of osteoclastogenesisp in rat periodontal ligament cells.

• Clinical and Experimental Reproductive Medicine
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• 제49권4호
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• pp.259-269
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• 2022

Objective: Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. Methods: Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. Results: The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. Conclusion: Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.

• 대한정형외과학회지
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• 제54권6호
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• pp.490-497
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• 2019

최근 줄기세포에 대한 생물학적 지식의 발전으로 인해 이를 실제 환자의 치료에 적용시키기 위한 다양한 노력들이 이루어지고 있다. 중간엽 줄기세포는 골수 흡인물로부터 처음 발견되었으나 현재는 지방, 피부, 근육, 제대혈 등 다양한 조직으로부터 추출될 수 있는 다능성 기질세포로 이해되고 있다. 그동안 중간엽 줄기세포의 골형성능은 여러 실험 및 동물 연구를 통해 증명되었으며 골결손, 골괴사, 불유합 등의 어려운 임상 상황에서 일부 성공적인 골재생 결과들이 보고되고 있다. 하지만 아직까지 각 질환별 적응증이나 표준화된 적용법이 마련되어 있지 않으며 효능 및 안전성에 대한 객관적 증거가 부족한 상태이다. 중간엽 줄기세포를 이용한 골재생은 앞으로 더욱 확대될 가능성이 높으나 표준적인 치료로 인정받기 위해서는 아직 해결되어야 할 과제들 또한 남아 있다.

• 대한한방내과학회지
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• 제30권2호
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• pp.327-337
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• 2009

Objective : In benign prostatic hyperplasia, dihydrotestosterone acts as a potent cellular androgen and promotes prostate growth. Inhibiting enzyme 5${\alpha}$-reductase, which is involved in the conversion of testosterone to the active form dihydrotestosterone, reduces excessive prostate growth. Recently Scutellaria baicalensis has been related reports about the effect of baicalein on anti-proliferation of the prostate gland. In this study, we investigated the effects of Scutellaria baicalensis on cytopathological alterations and expression of 5${\alpha}$-reductase in the rat model of benign prostatic hyperplasia induced by castration and testosterone treatment. Methods : Sprague-Dawley rat were treated with testosterone after castration for induction of experimental benign prostatic hyperplasia, which is similar to human benign prostatic hyperplasia in histopathological profiles. Scutellaria baicalensis as an experimental specimen, and finasteride as a positive control, were administered orally. The prostates were evaluated by histopathological changes, testosterone levels, and the expression of 5${\alpha}$-reductase genes. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Scutellaria baicalensis showed a diminished range of tissue damage. In the reverse transcription-polymerase chain reaction(RT-PCR) of 5${\alpha}$-reductase genes. Scutellaria baicalensis inhibited the expression of 5${\alpha}$-reductase genes. Conclusions : These findings suggest that Scutellaria baicalensis may protect glandular epithelial cells and also inhibit stromal proliferation in association with the suppression of 5${\alpha}$-reductase. From theses results, we suggest that Scutellaria baicalensis could be a useful remedy agent for treating benign prostatic hyperplasia.

• Nutrition Research and Practice
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• 제7권3호
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• pp.172-177
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• 2013

Benign prostatic hyperplasia (BPH) is one of the most common diseases among elderly men. As the old-age population is increasing recently, it is to our interest to observe the growing BPH within them. In BPH, the dihydrotestosterone (DHT) acts as promotes prostate growth. It inhibits enzyme $5{\alpha}$-reductase that is involved in the conversion of testosterone to the DHT activity which reduces the excessive prostate growth. Through experiments, the effects of Phellius linteus water extract performed on the BPH rats were induced by testosterone treatments. For 12 weeks, Sprague-Dawley rats were treated with testosterone for the induction of BPH. Rats were divided into four experimental groups: the not treated group (N), the testosterone injection and D.W treatment group (TN), the testosterone injection and Phellinus linteus treatment group (TP) and testosterone injection and finasteride treatment group (TF). Prostate weight, volume and weight ratio in the TP group and the TF group were significantly lower than the TN group. Testosterone and DHT levels in the TN group were significantly higher than that of the N group. And the TP group was significantly decreased than that of the TN group. While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation; the TP and TF groups showed trophic symptoms and were lined by flattened epithelial cells, thus, the stromal proliferation is relatively low as compared to the TN group. These suggest that Phellinus linteus water extracts may be an useful remedy for treating the benign prostatic hyperplasia.

• 한국한의학연구원논문집
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• 제16권2호
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• pp.131-141
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• 2010

Objective : Benign prostatic hyperplasia(BPH) is one of the most common diseased among elderly men. BPH can be treated with alpha-1 adrenergic blocker or $5{\alpha}$-reductase inhibitor(Finasteride) that reduces serum dihydrotestosterone(DHT). Phellodendri Cortex Ex has been broad studied on its chemical components, pharmacological activity, and clinical effects on anti-inflammation, anti-allergy, anti-tumor, immunity, antibacteria and other bioactivities. In this study, we investigated the therapeutic effects and action mechanism of Phellodendri Cortex Ex with a BPH induced by castration and testosterone treatment. Methods : Sprague-Dawley rats were treated with testosterone after castration for induction of experimental benign prostatic hyperplasia, which is similar to human benign prostatic hyperplasia in histopathological profiles. Phellodendri Cortex as an experimental specimen, and Finasteride as a positive control, were administered orally. The prostates were evaluated by histopathological changes, and the expression of $5{\alpha}$-reductase genes. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Phellodendri Cortex Ex showed a diminished range of the tissue damage. In the reverse transcription-polymerase chain reaction(RT-PCR) of $5{\alpha}$-reductase genes, Phellodendri Cortex inhibited the expression of $5{\alpha}$-reductase genes. Conclusions : These findings suggest that Phellodendri Cortex Ex may protect the glandular epithelial cells and also inhibit stromal proliferation in association with the suppression of $5{\alpha}$-reductase. From these results, we suggest that Phellodendri Cortex Ex could be a useful agent for treating the benign prostatic hyperplasia.

• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.93-100
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• 2004

많은 연구들에서 hMSC를 얻기 위해 centrifugation, fluoroscence activated cell sorter(FACS), magnetic activated cell sorter(MACS)가 이용되어져 왔다. 그러나 centrifugation만을 이용한 경우 순도가 떨어지며 FACS나 MACS의 경우에는 비용, 시간이 많이 드는 단점이 있다. 따라서 이 연구에서는 antibody cocktail을 이용하여 hMSC를 좀더 쉽게 얻어내는 방법에 대해 알아보았다. 사람의 골반에서 12G의 바늘을 이용하여 골수를 흡입한 후 heparin이 들어있는 시험관에 넣고 처리과정을 시행하기 전에 냉장고에 보관하며 가능한 한 빨리 처리 과정을 실시한다. 얻은 골수에 적당량의 RosetteSep( Stemcell Technologies)을 첨가한 후 실온에서 20분간 반응시킨다. 그 후 적당량의 Ficoll-paque위에 골수와 RosetteSep의 혼합물을 섞이지 않게 올리고 원심분리를 이용하여 원하는 세포층을 얻어낸다. 이 세포층을 따로 분리한 뒤 배양한다. 배양 시 세포가 80%이상 차기 전에 계속 passage를 시행하며 배양한다. 이는 세포가 밀도가 높아져 원치 않는 세포로 분화되는 것을 막기 위함이다. 배양된 세포가 다양한 분화능력을 가지고 있는지 알아보기 위해 세 가지로 분화를 유도하였다. 적절한 배지와 적절한 환경에서 배양함으로써 얻어진 세포를 osteoblast, chondroblast, adipocyte로 분화를 유도하였다. 분화된 세포가 원하는 형질의 세포로 분화되었는지를 확인하기 위하여 osteoblast의 경우 alizarin red staining, alkaline phosphatase activity, chondroblast의 경우 toluidine blue staining, adipocyte의 경우 Oil-Red-O staining으로 염색하여 분화를 확인하였다. 분리해낸 세포는 각각 세 가지 세포로 분화가 되었으며 이는 RosetteSep이 hMSC를 성공적으로 분리해냈다는 것을 보여준다. 그러나 모든 세포가 분화를 보이지는 않았으며 따라서 hMSC의 순도를 높이기 위한 연구가 더 필요하다. RosetteSep을 이용하면 다른 방법들 보다 쉽게 hMSC를 얻을 수 있으나 기존의 방법과 순도의 측면에서 더 비교할 필요가 있다.

• 대한한방내과학회지
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• 제31권1호
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• pp.166-176
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• 2010

Objective : Benign prostatic hyperplasia(BPH) is one of the most common diseases among elderly men. In BPH, dihydrotestosterone (DHT) acts as a potent cellular androgen and promotes prostate growth. Many reports conclude the component melittin in bee venom has the potential to treat various diseases including prostate cancer. In this study, we investigated the therapeutic effects and action mechanism of Bee venom herbal acupuncture with BPH induced by castration and testosterone treatment. Methods : Sprague-Dawley rats were treated with testosterone after castration for induction of experimental BPH. A total of 24 rats were equally divided into four groups: Group 1 was the model group; Group 2 served as control (sham-operated group); Group 3 animals were treated with Bee venom herbal acupuncture as an experimental specimen; Group 4 served as a positive control group and was treated with finasteride at a dose of 1 mg/kg. The drugs were administered orally. The prostates were evaluated by prostatic weight, volume, histopathological changes and testosterone levels. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Bee venom herbal acupuncture showed a diminished range of tissue damage and showed significant decrease in their prostatic weights, volume and histopathological examination. Conclusions : These results suggest that Bee venom herbal acupuncture may protect the glandular epithelial cells and also inhibit stromal proliferation. From theses results, we suggest that Bee venom herbal acupuncture could be a useful remedy agent for treating the benign prostatic hyperplasia.

• Clinical and Experimental Reproductive Medicine
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• 제40권2호
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• pp.67-75
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• 2013

Objective: To investigate the effect of peroxisome proliferator activated receptor ${\gamma}$(PPAR${\gamma}$) agonist on the cell proliferation properties and expression of human telomerase reverse transcriptase (hTERT) and aromatase in cultured endometrial stromal cell (ESC) from patients with endometriosis. Methods: Human endometrial tissues were obtained from women with endometriosis and healthy women (controls) using endometrial biopsy. Isolated ESCs were cultured and the cell proliferation was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay and expression of hTERT, aromatase, and cyclooxygenase (COX)-2 by western blotting according to the addition of rosiglitazone (PPAR${\gamma}$ agonist). Results: We demonstrate that the cultured ESCs of endometriosis showed hTERT protein overexpression and increased cellular proliferation, which was inhibited by rosiglitazone, in a dose-dependent manner. At the same time, PPAR${\gamma}$ agonist also inhibited aromatase and COX-2 expression, resulting in decreased prostaglandin $E_2$ production in the ESCs of endometriosis. Conclusion: This study suggests that PPAR${\gamma}$ agonist plays an inhibitory role in the proliferative properties of eutopic endometrium with endometriosis by down-regulation of hTERT and COX-2 expression; this could be a new treatment target for endometriosis.

• Parasites, Hosts and Diseases
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• 제49권3호
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• pp.235-243
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• 2011

In order to get a better understanding of the role of protease-activated receptor 2 (PAR2) in type 2 helper T (Th2) cell responses against Trichinella spiralis infection, we analyzed Th2 responses in T. spiralis-infected PAR2 knockout (KO) mice. The levels of the Th2 cell-secreted cytokines, IL-4, IL-5, and IL-13 were markedly reduced in the PAR2 KO mice as compared to the wild type mice following infection with T. spiralis. The serum levels of parasite-specific IgE increased significantly in the wild type mice as the result of T. spiralis infection, but this level was not significantly increased in PAR2 KO mice. The expression level of thymic stromal lymphopoietin, IL-25, and eotaxin gene (the genes were recently known as Th2 response initiators) of mouse intestinal epithelial cells were increased as the result of treatment with T. spiralis excretory-secretory proteins. However, the expression of these chemokine genes was inhibited by protease inhibitor treatments. In conclusion, PAR2 might involve in Th2 responses against T. spiralis infection.