Comparative analysis of secretomes from deciduous and permanent periodontal ligament cells

Dept. of Public Health/박사The aims of the present study were to identify and compare the secretomes of human deciduous and permanent periodontal ligament (PDL) cells. Secreted proteins were isolated and identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Antibodies against Tudor domain containing 7 (TDRD7) and Vasorin (VASN) were used for western blot. In addition, we used the cytokine membrane array analysis and enzyme-linked immunosorbent assay (ELISA) was performed for some cytokine (VEGF, EGF, PDGF-BB, SCF, and FGF beta). Collagen type VI, alpha I, collagen type I, alpha I, collagen type III, alpha I, cullin 7, and nuclear factor I/X (CCAAT-binding transcription factor) related to cell growth and/or maintenance were identified in deciduous PDL group. But vasorin (VASN), IGF binding protein 4, and S100 calcium binding protein A9 related to cell communication and signal transduction were identified in permanent PDL group. The secretomes that were upregulated in deciduous PDL tissues were involved in inflammatory or immune reactions (IL-1 alpha, IL-1 beta, IL-2, I-309, IL-12 p40/70, IL-4, MMP1). Proteins related to cell growth and/or maintenance, cell communication; signal transduction, inflammatory or immune reactions were slightly different between permanent PDL and deciduous PDL groups. These results show that humoral factors released from deciduous and permanent PDL cells have different characteristics. Although the findings in our study are not sufficient to explain difference between permanent and deciduous PDL clearly, our study provides clues to develop our understanding of the molecular basis.ope

건조 조건에 따른 의류소재의 변형 연구 및 최적 건조 조건 제안

학위논문 (석사)-- 서울대학교 대학원 생활과학대학 의류학과, 2017. 8. 박정희.본 연구는 다양한 의류소재에 대해 건조 조건에 따른 변형을 측정하여 섬유 별로 건조 과정에서 변형을 초래하는 영향요인을 분석하고 건조 조건 별 효율적 건조 조건을 제안하고자 하였다. 이를 위하여, 섬유의 특성과 조직이 다른 면, 양모, 라이오셀, 나일론/스판덱스, 폴리에스터를 시료로 선정하고 온도, 습도, 건조 방식이 다른 네 가지 건조 조건을 설정하였다. 건조 조건에 따라 구김, 길이 변화율, 면적 변화율 및 건조 효과를 살펴보았으며, 최종적으로는 건조 조건 별로 최종 건조 효과와 변형을 유지하면서 시간과 에너지가 절약되는 최적 건조 조건을 제안하였다. 의류소재의 건조 후 변형은 조직 별로는 실의 자유도가 적은 직물이 편물보다 구김이 심했지만 수축은 적게 발생했다. 섬유의 화학적 성질 별로는 친수성 소재의 건조 후 구김과 수축은 건조 중 평균 온도에 영향을 받지 않았고, 건조 중 평균 습도가 낮을수록 구김이 심화되며 수축은 영향력이 미미하였다. 소수성 소재는 건조 중 평균 온도가 높을수록 수축이 심화되었고 구김에는 영향을 미치지 않았다. 또한 건조 중 평균 습도가 소수성 소재의 구김과 수축에 미치는 영향은 미미하였다. 기계력의 영향을 보면, 기계력이 가해지는 Heater, Heatpump(t) 조건에서 수축은 컸지만 텀블링에 의하여 세탁에 의한 구김이 일부 펼쳐졌으며, 기계력이 없는 Heatpump(h), Line Dry 조건에서는 시료의 수축은 적었지만 구김이 심화되는 경향을 나타내었다. 특히, 양모 시료에서 기계력의 영향이 크게 나타났으며 이는 양모가 기계력에 의해 축융 수축하였기 때문으로 사료된다. 반복 건조 후 의류소재는 전반적으로 최초 건조 시 보다 형태 및 치수 안정성을 나타내어 최초 건조 시 변형을 최소화할 수 있는 건조 조건을 선택하는 것이 필요함을 알 수 있었다. 예외적으로, 양모는 지속적인 세탁과 건조에 의해 축융 수축하여 구김과 수축이 심화되어 의류 관리 시 각별한 주의가 필요함을 시사한다. 본 연구에서는 모든 건조 조건에서 건조 중 표준수분율 이하에 도달한 뒤 길이변화율에 변화가 미미한 구간을 발견하였다. 이에 이 구간을 과잉건조되는 구간으로 판단하고, 건조 조건 별 적정 건조 시간을 세탁부하직물 1kg을 기준 Heater 조건 30분, Heatpump(t) 조건 50분, Heatpump(h) 조건 30분으로 설정하였다.Ⅰ. 서 론 1 1. 연구의 필요성 및 목적 1 2. 이론적 배경 4 2.1. 섬유의 수분 흡수와 건조 4 2.2. 의류소재의 물리적 변형 7 Ⅱ. 실 험 12 1. 시료 12 2. 세탁 14 3. 건조 15 3.1. 건조 조건 15 3.2. 건조 방법 18 3.3. 건조기 내 온·습도 환경 20 4. 건조효과 및 변형 평가 21 4.1. 길이 및 면적 변화율 21 4.2. 구김 평가 23 4.3. 수분율 25 Ⅲ. 결과 및 고찰 26 1. 건조 조건에 따른 의류소재의 형태 변형 26 1.1. 건조 조건 별 건조 중 온 · 습도 환경 26 1.2. 구김 특성 29 1.3. 수축 특성 32 2. 건조 조건이 의류소재 변형에 미치는 영향 42 2.1. 온 · 습도의 영향 43 2.2. 기계력의 영향 48 2.3. 반복 건조의 영향 51 3. 효율적 건조 조건 57 3.1. 수분율 감소에 따른 수축 특성 58 3.2. 효율적 건조 조건의 제안 65 Ⅳ. 요약 및 결론 68 Ⅴ. 참고 문헌 72 Abstract 76Maste

The Inhibitory Effect of Buddlejasaponin IV on the Growth of YD-10B Human Oral Squamous Cell Carcinoma Cells

Background: Buddlejasaponin IV (BS-IV), a triterpene saponin isolated from Pleurospermum kamtschaticum HOFFMANN (Umbelliferae), is known to have potent anti-inflammatory activity and cytotoxicity against diverse cancer cell lines. In the present study, we attempted to verify whether BS-IV could inhibit cell growth, and induce cell cycle arrest and apoptosis in highly invasive YD-10B human oral squamous cell carcinoma (OSCC) cells. Methods: YD-10B cells were treated with various concentrations of BS-IV, and the cell viability was evaluated by MTT assay. Flow cytometry was conducted to examine cell phase distribution and DAPI staining was performed to observe apoptotic morphological changes in BS-IV-treated YD-10B cells. Western blot analysis was used to investigate the expression of proteins associated with cell cycle arrest and apoptosis.Results: BS-IV treatment significantly reduced the viability of YD-10B cells and partially arrested cell cycle progression at the G2/M phase. Treatment with BS-IV substantially decreased the levels of cyclin B1 and stimulated the phosphorylation of checkpoint kinase 2 (Chk2). The expression of p21 was increased but the phosphorylation of Akt was inhibited in BS-IV-treated YD-10B cells. Furthermore, BS-IV induced release of cytochrome c from mitochondria by reducing anti-apoptotic Bcl-2 level and increasing pro-apoptotic Bax level. Active caspase-3 level and the cleavage of poly (ADP-ribose) polymerase (PARP) were enhanced by BS-IV treatment. In addition, BS-IV increased the expression of Fas death receptor and its ligand (FasL) in YD-10B cells. Conclusions: The treatment with BS-IV inhibits the growth of YD-10B cells by inducing p21-dependent cell cycle arrest at G2/M phase and apoptosis through both mitochondrial-dependent and death receptor-mediated pathways. Thus, BS-IV is an excellent candidate for a chemopreventive agent to block the progression of human OSCC.ope

Synthesis and Anticancer Activity of Novel Deoxoartemisinin-Glycolipid Hybrids

The practical synthesis and anticancer activity of novel deoxoartemisinin-glycolipid hybrids, which incorporate two drugs into a single molecule and can impact multiple targets simultaneously are presented. These hybrids exhibited potent in vitro anticancer activity against several human cancer cell lines. The deoxoartemisinin-glycolipid hybrids generally demonstrated better anticancer activity than either artemisinin or daumone alone and cisplatin.ope

Honokiol inhibits the progression of collagen-induced arthritis by reducing levels of pro-inflammatory cytokines and matrix metalloproteinases and blocking oxidative tissue damage.

Plant-derived compounds with potent anti-inflammatory activity have attracted a great deal of attention as a source for novel anti-arthritic agents with minimal side effects. We attempted to determine the anti-arthritic effects of orally administered honokiol isolated from Magnolia species. The oral administration of honokiol inhibited the progression and severity of type II collagen (CII)-induced arthritis (CIA) by reducing clinical arthritis scores and paw swelling. The histological analysis demonstrated preserved joint space; and the immunohistochemical data showed that the levels of interleukin (IL)-17, matrix metalloproteinase (MMP)-3, MMP-9, MMP-13, and receptor activator for nuclear factor-κB ligand, as well as nitrotyrosine formation, were substantially suppressed in the honokiol-treated CIA mice. The elevated serum levels of tumor necrosis factor-α and IL-1β in the CIA mice were also restored to control levels via honokiol treatment. In the CIA mice, honokiol inhibited CII- or lipopolysaccharide-stimulated cytokine secretion in spleen cells, as well as CII-stimulated spleen cell proliferation. Furthermore, honokiol treatment reduced CIA-induced oxidative damage in the liver and kidney tissues of CIA mice. Collectively, the oral administration of honokiol inhibited CIA development by reducing the production of pro-inflammatory cytokines, MMP expressions, and oxidative stress. Thus, honokiol is an attractive candidate for an anti-arthritic agent.ope

Red ginseng saponin extract attenuates murine collagen-induced arthritis by reducing pro-inflammatory responses and matrix metalloproteinase-3 expression.

Ginseng, the root of Panax ginseng C. A. MEYER, has been used as a food product and medicinal ingredient. In this study, we assessed the anti-arthritic effects of red ginseng saponin extract (RGSE), including ginsenosides Rg3, Rk1 and Rg5 as major components, on a murine type II collagen (CII)-induced arthritis (CIA), which is a valid animal model of human arthritis. Oral administration of RGSE at 10 mg/kg reduced the clinical arthritis score and paw swelling in the CIA mice, and inhibited joint space narrowing and histological arthritis, illustrating the severity of synovial hyperplasia, inflammatory cell infiltration, pannus formation, and erosion of cartilage. RGSE inhibited the expression of matrix metalloproteinase-3 and nitrotyrosine formation, and recovered the expression of superoxide dismutase in the joints of the CIA mice. Orally administered RGSE also reduced the levels of serum tumor necrosis factor-alpha and interleukin-1beta in the CIA mice. CII- or lipopolysaccharide-stimulated cytokine production, in addition to CII-specific proliferation, was reduced in the spleen cells of the RGSE-treated CIA mice, as compared with those from vehicle-treated CIA mice. Furthermore, RGSE administration protected against CIA-induced oxidative tissue damage by restoring the increased malondialdehyde levels and the decreased glutathione levels and catalase activities almost to control levels. Therefore, RGSE may be a beneficial supplement which can improve human arthritis.ope

Comparative analysis of secretory factors from permanent- and deciduous-teeth periodontal ligament cells

OBJECTIVE: Studies of regenerative therapies have focused on the paracrine effects of mesenchymal stem cells, but little has been revealed about the humoral factors of periodontal ligament (PDL) stem cells. The aim of this study was to identify and compare the secretory factors of human permanent- and deciduous-teeth PDL cells (P-PDL and D-PDL cells, respectively) in order to understand the characteristics of these cells and their potential applications in regenerative therapies. DESIGN: Conditioned media were collected from P-PDL and D-PDL cells (P-PDL-CM and D-PDL-CM, respectively). These media were analyzed with high-performance liquid-chromatography-coupled electrospray ionization tandem mass spectrometry and a cytokine membrane assay. In addition, Western blot analysis was performed to verify the differences between the two media. RESULTS: Cytokines related to neurogenesis (NT-3 and NT-4) and angiogenesis-related cytokines (EGF and IGF-1) were identified in P-PDL-CM. The expression levels of immune-response-related cytokines (interleukins I, II, and IV) and secreted proteins related to tissue degradation and catalytic activities (matrix metallopeptidase 1 (MMP1), Proteasome subunit, alpha type, 1 (PSMA1), and cullin 7 (CUL7)) were higher in D-PDL-CM. Vasorin (VASN) was expressed more strongly in P-PDL-CM, but tudor domain containing 7 (TDRD7) was expressed more strongly in D-PDL-CM in Western blot analysis. CONCLUSION: The cytokine expressions of the two cell types showed different patterns, especially in neurogenesis and immune responses. P-PDL cells are more suitable candidates for applications in regenerative therapies.restrictio

Betulinic acid, a bioactive pentacyclic triterpenoid, inhibits skeletal-related events induced by breast cancer bone metastases and treatment

Many breast cancer patients experience bone metastases and suffer skeletal complications. The present study provides evidence on the protective and therapeutic potential of betulinic acid on cancer-associated bone diseases. Betulinic acid is a naturally occurring triterpenoid with the beneficial activity to limit the progression and severity of cancer, diabetes, cardiovascular diseases, atherosclerosis, and obesity. We first investigated its effect on breast cancer cells, osteoblastic cells, and osteoclasts in the vicious cycle of osteolytic bone metastasis. Betulinic acid reduced cell viability and the production of parathyroid hormone-related protein (PTHrP), a major osteolytic factor, in MDA-MB-231 human metastatic breast cancer cells stimulated with or without tumor growth factor-β. Betulinic acid blocked an increase in the receptor activator of nuclear factor-kappa B ligand (RANKL)/osteoprotegerin ratio by downregulating RANKL protein expression in PTHrP-treated human osteoblastic cells. In addition, betulinic acid inhibited RANKL-induced osteoclastogenesis in murine bone marrow macrophages and decreased the production of resorbed area in plates with a bone biomimetic synthetic surface by suppressing the secretion of matrix metalloproteinase (MMP)-2, MMP-9, and cathepsin K in RANKL-induced osteoclasts. Furthermore, oral administration of betulinic acid inhibited bone loss in mice intra-tibially inoculated with breast cancer cells and in ovariectomized mice causing estrogen deprivation, as supported by the restored bone morphometric parameters and serum bone turnover markers. Taken together, these findings suggest that betulinic acid may have the potential to prevent bone loss in patients with bone metastases and cancer treatment-induced estrogen deficiency.ope

Platycodin D Blocks Breast Cancer-Induced Bone Destruction by Inhibiting Osteoclastogenesis and the Growth of Breast Cancer Cells

BACKGROUND: Metastatic breast cancer cells are frequently associated with osteoclast-mediated bone resorption, resulting in severe bone destruction and increased mortality in patients. Platycodin D (PD) isolated from Platycodon grandiflorum is a triterpenoid saponin with anti-cancer and anti-angiogenic potential. METHODS: The in vivo activity was determined in mice with the intratibial injection of human metastatic breast cancer cells. Osteoclast formation and activity were detected using tartrate-resistant acid phosphatase staining and calcium phosphate-coated plates. The expression of osteoclastogenesis-inducing molecules was detected by RT-PCR and western blotting in RANKL-treated bone marrow macrophages (BMMs). Cell viability and DNA synthesis were measured with MTT and BrdU incorporation assays. The induction of apoptosis was estimated using TUNEL staining and a caspase-3 activity assay. RESULTS: The oral administration of PD inhibited MDA-MB-231 cell-induced osteolysis in an intratibial mouse model. PD treatment blocked RANKL-induced osteoclast formation by inhibiting the expression and nuclear translocation of NFATc1 and c-Fos in BMMs and consequently reduced osteoclast-mediated bone resorption. Furthermore, PD treatment induced apoptosis in osteoclasts and inhibited the growth of MDA-MB-231 cells. CONCLUSION: PD may block breast cancer-induced bone loss by suppressing the formation, activity, and survival of osteoclasts, as well as the growth of metastatic breast cancer cells.ope