• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8533-8539
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• 2016

Background: B-cell chronic lymphocytic leukemia B (B-CLL), the most common type of leukemia, may be caused by apoptosis deficiency in the body. Adipose tissue-derived mesenchymal stem cells (AD-MSCs) as providers of pro-apoptotic molecules such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), can be considered as an effective anti-cancer therapy candidate. Therefore, in this study we assessed the role of tumor necrosis factor-producing mesenchymal stem cells oin apoptosis of B-CLL cells resistant to fludarabine-based chemotherapy. Materials and Methods: In this study, after isolation and culture of AD-MSCs, a lentiviral LeGO-iG2-TRAIL-GFP vector containing a gene producing the ligand pro-apoptotic with plasmid PsPAX2 and PMDG2 virus were transfected into cell-lines to generate T293HEK. Then, T293HEK cell supernatant containing the virus produced after 48 and 72 hours was collected, and these viruses were transduced to reprogram AD-MSCs. Apoptosis rates were separately studied in four groups: group 1, AD-MSCs-TRAIL; group 2, AD-MSCs-GFP; group 3, AD-MSCs; and group 4, CLL. Results: Observed apoptosis rates were: group 1, $42{\pm}1.04%$; group 2, $21{\pm}0.57%$; group 3, $19{\pm}2.6%$; and group 4, % $0.01{\pm}0.01$. The highest rate of apoptosis thus occurred ingroup 1 (transduced TRAIL encoding vector). In this group, the average medium-soluble TRAIL was 72.7pg/m and flow cytometry analysis showed a pro-apoptosis rate of $63{\pm}1.6%$, which was again higher than in other groups. Conclusions: In this study we have shown that tumor necrosis factor (TNF) secreted by AD-MSCs may play an effective role in inducing B-CLL cell apoptosis.

• Journal of Periodontal and Implant Science
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• 제39권sup2호
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3595-3604
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• 2015

Hepatocellular carcinoma (HCC) has been one of the most fatal malignant tumors worldwide and its associated morbidity and mortality remain of significant concern. Based on in-depth reviews of serological diagnosis of HCC, in addition to AFP, there are other biomarkers: Lens culinaris agglutinin-reactive AFP (AFP-L3), descarboxyprothrombin (DCP), tyrosine kinase with Ig and eprdermal growth factor (EGF) homology domains 2 (TIE2)-espressing monocytes (TEMs), glypican-3 (GPC3), Golgi protein 73 (GP73), interleukin-6 (IL-6), and squamous cell carcinoma antigen (SCCA) have been proposed as biomarkers for the early detection of HCC. The diagnosis of HCC is primarily based on noninvasive standard imaging methods, such as ultrasound (US), dynamic multiphasic multidetector-row CT (MDCT) and magnetic resonance imaging (MRI). Some experts advocate gadolinium diethyl-enetriamine pentaacetic acid (Gd-EOB-DTPA) MRI and contrast-enhanced US as the promising imaging madalities of choice. With regard to recent advancements in tissue markers, many cuting-edge technologies using genome-wide DNA microarrays, qRT-PCR, and proteomic and inmunostaining studies have been implemented in an attempt to identify markers for early diagnosis of HCC. Only less than half of HCC patients at initial diagnosis are at an early stage treatable with curative options: local ablation, surgical resection, or liver transplant. Transarterial chemoembolization (TACE) is considered the standard of care with palliation for intermediate stage HCC. Recent innovative procedures using drug-eluting-beads and radioembolization using Yttrium-90 may exhibit beneficial effects in HCC treatment. During the past few years, several molecular targeted agents have been evaluated in clinical trials in advanced HCC. Sorafenib is currently the only approved systemic treatment for HCC. It has been approved for the therapy of asymptomatic HCC patients with well-preserved liver function who are not candidates for potentially curative treatments, such as surgical resection or liver transplantation. In the USA, Europe and particularly Japan, hepatitis C virus (HCV) related HCC accounts for most liver cancer, as compared with Asia-Pacific regions, where hepatitis B virus (HBV) may play a more important role in HCC development. HBV vaccination, while a vaccine is not yet available against HCV, has been recognized as a best primary prevention method for HBV-related HCC, although in patients already infected with HBV or HCV, secondary prevention with antiviral therapy is still a reasonable strategy. In addition to HBV and HCV, attention should be paid to other relevant HCC risk factors, including nonalcoholic fatty liver disease due to obesity and diabetes, heavy alcohol consumption, and prolonged aflatoxin exposure. Interestingly, coffee and vitamin K2 have been proven to provide protective effects against HCC. Regarding tertiary prevention of HCC recurrence after surgical resection, addition of antiviral treatment has proven to be a rational strategy.

• 공업화학
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• 제32권3호
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• pp.312-319
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• 2021

광열 치료(photothermal therapy)란 빛을 조사하여 열을 발생시킴으로써 정상세포보다 열에 약한 비정상 세포, 특히 암세포를 선택적으로 괴사시키는 치료법이다. 본 연구에서는 광열 치료를 위한 카르복실화된 환원 그래핀옥사이드(reduced graphene oxide with carboxyl groups, CRGO)-골드나노막대(gold nanorod, AuNR) 나노복합체를 합성하고자 하였다. 이를 위해 그래핀옥사이드(graphene oxide, GO)를 고온에서 선택적으로 환원, 박리하여 CRGO를 합성하였고, AgNO₃의 양에 따라 AuNR의 길이를 조절하여 880 nm에서 강한 흡광 특성을 나타내는 AuNR를 합성하여 광열 인자로 사용하였다. 일반적인 방법으로 환원된 RGO에 비해 CRGO에 상대적으로 많은 카르복실기가 결합되어 있음을 FT-IR, 열 중량 분석 및 형광 분석을 통해 확인하였다. 또한, RGO에 비해 많은 carboxyl group이 결합된 CRGO는 수용액상에서 우수한 안정성을 나타내었다. 정전기적 상호작용을 통해 합성된 CRGO-AuNR 나노복합체는 약 317 nm의 균일한 크기와 좁은 크기 분포를 보였다. CRGO-AuNR 나노복합체는 두 가지 광열 인자인 CRGO와 AuNR의 synergistic effect로 인하여 조직 투과도가 우수한 근적외선 880 nm 레이저의 조사에 의한 광열 효과가 AuNR보다 2배 이상 향상 되는 것을 확인하였다. 또한, 광열 효과에 의한 암세포 독성 분석 결과, CRGO-AuNR 나노복합체가 가장 우수한 세포 독성 특성을 나타내었다. 따라서 CRGO-AuNR 나노복합체는 안정된 분산성과 향상된 광열 효과를 기반으로 항암 광열 요법 분야에 응용될 수 있을 것으로 기대된다.

• 생약학회지
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• 제46권1호
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• pp.52-58
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• 2015

Neural stem cells (NSCs), with self-renewal and neuronal differentiation capacity, are a feasible resource in cell-based therapies for various neurodegenerative diseases and neural tissue injuries. In this study, we investigated the effects of Schisandrae Fructus (SF) on proliferation and differentiation of human embryonic NSCs. Treatment with 70% ethanol extract of SF increased the viability of NSCs derived from human embryonic stem cells, which was accompanied by increased mRNA expression of cyclin D1. Whereas 70% ethanol extract of SF also decreased the mRNA expression of nestin, it increased class III ${\beta}$-tublin (Tuj-1) and MAP2 in both growth and differentiation media. Lastly, we found increased mRNA expression of BDNF in SF-treated NSCs. In conclusion, our study demonstrates for the first time that SF induced proliferation and neuronal differentiation of NSCs and increased mRNA expression of BDNF, suggesting its potential as a regulator of NSC fate in NSC-based therapy for neuronal injuries from various diseases.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권2호
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• pp.111-117
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• 2006

An area of current research is investigating the app1ication of human mesenchymal stem cells or hMSCs as a cell-based regenerative therapy. In order to achieve effective bone regeneration, appropriate matrices functioning as cell-carriers must be identified and optimized in terms of function, efficacy and biocompatibility. Two methods of approaching optimization of matrices are to facilitate adhesion of the donor hMSCs and furthermore to facilitate recruitment of host progenitor cells to osteoblastic differentiation. Pleiotrophin is an extracellular matrix protein that was first identified in developing rat brains and believed to be associated with developing neuronal pathways. A recent publication by Imai and colleagues demonstrated that transgenic mice with upregulated pleiotrophin expression developed a greater volume of cortical as well as cancellous bone. The proposed mechanism of action of pleiotrophin is demonstrated here. Through either environmental stresses and/or intracellular regulation, there is an increase in pleiotrophin production. The pleiotrophin is released extracellularly into areas requiring bone deposition. A receptor-mediated process recruits host osteoprogenitor cells into these areas. Therefore, the aim of our study was to investigate the osteoconductive properties of pleiotrophin. We wanted to determine if pleiotrophin coating facilitates cellular adhesion and furthermore if this has any effect on hMSCs derived bone formation in an animal model. The results showed a dose dependent response of cellular adhesion in fibronectin samples, and cellular adhesion was facilitated with increasing pleiotrophin concentrations. Histologic findings taken after 5 weeks implantation in SCID mouse showed no presence of bone formation with only a dense fibrous connective tissue. Possible explanations for the results of the osteogenesis assay include inappropriate cell loading.

• 대한본초학회지
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• 제27권6호
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• pp.123-129
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• 2012

Objectives : Allergy has been described as an inflammatory with hypersensitivity resulting from seasonal or perennial responses to specific allergens. The root of Platycodon grandiflorum (Jacq.) A. DC.(Platycodi Radix; Campanulaceae) has been traditionally used to treat chronic diseases such as bronchitis, asthma, pulmonary tuberculosis, inflammation and hyperlipidemia. In this study, we examined the effect of 70% ethanol extract of Platycodi Radix (PR-E) on ovalbumin (OVA)-induced airway inflammation in mice. Methods : Mice were sensitized and challenged by OVA inhalation to induced chronic airway inflammation, and then were intragastrically administered PR-E extract at doses of 50 and 200 mg/kg/day from days 21 to 30 consecutively. The levels of allergic mediators such as histamine, OVA-specific immunoglobulin (Ig) E, and cytokines such as IL-4 and IFN-${\gamma}$ were measured in the sera of mice by ELISA. The histological change of lung tissue was observed by hematoxylin and eosin (H&E) staining. Results : PR-E extract significantly decreased the serum levels of histamine, OVA-specific IgE, and Th2 cytokine, IL-4 compared with those in the OVA-induced group. PE-E extract significantly increased the serum level of Th1 cytokine, IFN-${\gamma}$. Based on lung histopathological studies, inflammatory cell infiltration and mucus hypersecretion were inhibited by PE-E extract administration compared to that in the OVA-induced group. Conclusions : These findings indicate that PE-E extract may be useful as an adjuvant therapy for the treatment of bronchial asthma.

• 대한화상학회지
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• 제22권2호
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• pp.45-48
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• 2019

Allogenic keratinocyte application is widely used for treatment of second degree burns. However, there is no significant body of report on application of allogenic keratinocyte to third degree burns. A geriatric patient visited our burn center showing second to third-degree burn on dorsum of her left hand. Considering the surface area and wound depth, surgery was indicated but her medical condition and age made the surgery high risk for a long operation. Therefore, chemical escharolysis, serial bedside debridement, and cultured allogenic keratinocyte (Kaloderm^®, Tegoscience, Seoul, Korea) application was done. The wound was completely epithelialized after four rounds of Kaloderm^® application. For third-degree burns where definitive surgical reconstruction is precluded due to medical comorbidity of the patient, we investigated the possibility of allogenic keratinocyte treatment which may allow to avoid high-risk anesthesia and surgery.

• Journal of Periodontal and Implant Science
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• 제33권3호
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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.

• Journal of Korean Neurosurgical Society
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• 제30권3호
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• pp.263-271
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• 2001

교모세포종은 비교적 흔한 원발성 뇌종양이며 생물학적 특성상 빠른 성장률을 보이는 것 외에 침습성이 강하여 종양과 인접한 부분을 파괴 시킬 뿐 아니라 직접접촉하지 않는 부분의 파괴도 일어나게 되어 그 결과 치료 예후가 매우 불량한 것으로 되어 있다. 이러한 불량한 예후를 개선 시키기 위해서는 이들 종양의 침습에 대한 기전의 정확한 이해가 필요하며 이를 이용한 새로운 치료방법이 요구된다할 것이다. Protein kinase C(PKC)는 세포내 신호전달체제 과정에서 매우 중요한 역할을 하는 효소로 세포막 수용체 신호를 핵으로 전달하는 역할을 하며 세포내 여러 생물학적 작용이 알려져 있다. 본 실험은 종양침습과 연관하여 세포내 PKC가 어떠한 작용을 하는지에 대해서 악성교종 세포를 대상으로 하여 알아보고자 하였다. 따라서 PKC가 종양침습에 중요한 역할을 할 것이라는 가설을 세웠고 이 가설을 증명하기 위해 세포내 PKC농도를 길항제 및 촉진제를 이용하며 높고 낮게 조절함으로써 그에 따른 침습성의 변화를 살펴보았다. 방법으로는 교모세포종 세포주인 U-87 세포를 약제로 처리한 후 인위적으로 조절된 세포내의 PKC에 대해 효소의 활성도를 측정하였고 침습성은 matrigel artificial basement membrane assay 및 tumor spheroid fetal rat brain aggregate(FRBA) confrontation assay를 이용하여 측정하였다. 결과로 PKC의 길항제인 tamoxifen과 hypericin으로 처치한 세포는 PKC의 활성과 침습도가 모두 감소하였으며 이는 약제농도에 비례하여 나타났다. 반면 PKC 자극제인 TPA로 처치된 세포는 증가된 PKC 활성도나 침습도을 보이지 않았다. 이러한 결과를 종합해 보았을 때 PKC는 종양세포의 침습성에 중요한 역할을 함을 알 수 있었으며 PKC의 길항제는 종양 치료에 유용한 화학 요법 제가 될 수 있을 것으로 사료된다.