• Korean Journal of Acupuncture
• /
• 제32권2호
• /
• pp.51-58
• /
• 2015

Objectives : Pinus densiflora gnarl, called Song-Jeol in Korean medicine, has been used to cure inflammatory diseases such as arthritis. In the present study, we evaluated inhibitory property of Song-Jeol pharmacopuncture(SJ) on C6 glioma cell migration. Methods : To evaluate cell viability on C6 glioma cells of SJ, the viability was assessed by using Ez-cytox assay kit. The cell migration was assessed by wound-healing assay and Boyden chamber assay, respectively. LPS-induced NO productions were determined by using the Griess reagent. The expression of iNOS and protein kinase $C(PKC)-{\alpha}$ were estimated by western blotting assay. Results : In the wound-healing assay and Boyden chamber assay, SJ showed a significant inhibition on serum-induced C6 glioma cell migration. In addition, NO production was decreased by SJ through suppression of iNOS expression in LPS-stimulated C6 glioma cell. Futhermore, LPS-induced protein kinase $C(PKC)-{\alpha}$ expression was effectively inhibited by SJ. Conclusions : These results demonstrated that SJ was useful for the suppression of the C6 glioma cell migration.

• 한국임상수의학회지
• /
• 제30권6호
• /
• pp.403-408
• /
• 2013

본 연구의 목적은 in vitro 실험에서 human dermal fibroblast cell을 이용한 섬유아세포 증식과 콜라겐 생성을 통한 전층피부창상치유에 대한 효과를 알아보는 것이다. In vitro 실험에서, 베타글루칸 및 양성대조물질 oncostatin M을 사용하여 처치 후 24 또는 48시간 이후 MTT assay와 세포수 관찰을 통해 섬유모세포 증식효과에 대해 평가하였다. Procollagen 생산에 대한 효과를 평가하기 위해 베타글루칸 및 양성대조물질 oncostatin M 처치 48시간 후 hydroxyproline을 HPLC를 이용하여 정량하여 평가하였다. 창상재구축에 대한 효과를 평가하기 위해 베타글루칸 및 양성대조물질 fucoidan fraction 7을 FPCL 모델을 이용하여 평가하였다. 베타글루칸 처치군은 음성대조군 대비 0.1 mg/ml 이상의 농도에서 용량의존적이고 유의적인 OD 또는 섬유아세포수의 증가가 관찰되었다. 게다가 베타글루칸 처치군은 창상면으로 유주된 세포수가 음성대조군 대비 1 mg/ml 이상의 농도에서 용량의존적이고 유의적인 증가가 관찰되었다. 그러나 베타글루칸 처치군은 음성대조군 대비 모든 용량에서 procollagen 생산에 대한 유의적인 변화가 관찰되지 않았다.

• Restorative Dentistry and Endodontics
• /
• 제45권1호
• /
• pp.2.1-2.7
• /
• 2020

Objectives: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. Materials and Methods: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethylthiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). Results: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). Conclusions: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

• 농약과학회지
• /
• 제9권4호
• /
• pp.429-436
• /
• 2005

오이에 균핵병을 일으키는 Sclerotinia sclerotiorum은 $25^{\circ}C$에서 균사의 생육이 가장 왕성하였으며, 병원성 실험을 위해 오이 열매를 이용한 실내 검정 실험의 경우 동일한 온도에서 병원성이 가장 강한 것으로 나타났다. 분리한 모든 Sclerotinia sclerotiorum 균주는 오이 절편 접종법과 상처 접종법을 사용한 실내 검정에서 모두 강한 병원성을 나타냈지만, 오이 절편 접종법의 경우는 상처 접종법과는 다르게, 균주와 환경, 처리한 살균제 간에 차이를 볼 수 없을 정도로 병원성이 강하여 실내 검정법으로는 적합하지 못하였다. 오이 절편접종법과 상처접종법을 사용하여 dichlofluanid와 carbendazim과 diethofencarb의 혼합제의 병 발생 억제 효과를 조사한 결과, 오이 절편접종법에서는 500과 $3.0{\mu}g/mL$의 처리구에서 14.3과 42.8%의 낮은 효과를 보였지만, 상처 접종법에서는 동일한 농도에서 100와 92.5%의 우수한 발병 억제 효과를 보였다. 두 살균제는 포장에서도 우수한 효과를 보였는데, $100{\mu}g/mL$의 혼합제와 $825{\mu}g/mL$의 dichlofluanid는 병 발생 억제 효과가 각각 91.1과 82.9%이었다. 충남 연기 지역의 오이 시설 재배지에서 분리한 37개의 S. sclerotioum 균주는 7가지의 살균제에 대한 저항성 모니터링을 실시하여 균주들의 평균 EC50값을 구하였는데, fenhexamid는 $0.13{\mu}g/mL$, 병원균의 세포막에서 과산화반응을 일으켜 병원균을 사멸시키는 것으로 알려진 procymidon과 iprodione은 0.18과 $0.24{\mu}g/mL$, Tubulin 단백질의 중합 반응을 억제하는 것으로 보고된 carbendazim과 carbendazim과 diethofencarb의 혼합제는 0.13과 $0.05{\mu}g/mL$, 보호용 살균제로 알려진 iminoctadine과 dichlofluanid는 평균 $EC_{50}$ 값이 1.94와 $8.95{\mu}g/mL$이었다. 특별히 dichlofluanid의 경우는 $EC_{50}$값의 범위가 0.001에서 $21.54{\mu}g/mL$ 사이의 넓은 범위에 분포하였지만, 포장에서의 사용 농도가 $1,650{\mu}g/mL$인 것을 감안한다면 저항성균의 출현은 확인하지 못하였다. 충북의 연기 지역에서는 7가지 살균제에 대하여 저항성을 뚜렷하게 나타내는 균핵병균은 발견하지 못하였다.

• 약학회지
• /
• 제37권5호
• /
• pp.532-537
• /
• 1993

Angiogenesis refers to the formation of new capillary blood vessels, or neovascularization occurring under various physical conditions, such as development of the embryo, formation of corpus luteum, wound healing and pathological conditions including tumor growth and metastases, hemangiomas, diabetic retinopathy, rheumatoid arthritis. There are many evidences that angiogenesis is important for the progressive growth of solid tumors and also permits the shedding of metastatic tumors from the primary site. Thus, treatment of angiogenesis inhibitors might be a novel strategy for tumor growth inhibition. Normal vascular endothelial cells are in a state of differentiation and angiogenic endothelial cells migrate and proliferate, and they subsequently differentiate into vessel-forming quiescent phenotype cells, Therfore, it was speculated that a modifier of cell differentiation could also affect angiogenesis. In order to identify new antiangiogenic factors, the research was conducted to estimate the inhibitory activities of cell differentiation agents by means of chick embryo chorioallantoic membrane(CAM) assay. Hence, we have established the CAM assay for the screening of antiangiogenic agents. Using the CAM assay, we found that ursolic acid, a tumor cell differentiation-inducing agent, showed a markedly inhibitory effect on chick embryonic angiogenesis.

• Archives of Pharmacal Research
• /
• 제26권10호
• /
• pp.855-860
• /
• 2003

The effect of silver sulfadiazine (SSD) on the proliferation of human dermal fibroblast (HDF) was studied to determine the impact of the drug on the wound healing process and dermal mechanical strength. Human dermal fibroblasts were cultured to 80% confluency using DMEM with 10% FBS and viability of the cell was estimated using neutral red assay. In addition, the $2^{nd}$ degree burn wound was prepared on the anterior part of rabbit ear skin and dressings containing SSD were applied for 96 h. Presence of inflammatory cells and degree of re-epithelialization were investigated in the wound. After 15 day of the induction of burn wounds, the treated area was excised and dermal mechanical strength was quantitatively measured with a constant speed tensiometer. SSD was found to be highly cyto-toxic in cultured HDF cells. The topical application of SSD (2%) could control the infection as evidenced by the lack of accumulation of inflammatory cells in histological evaluation. Therefore, these observations suggested that the impairment of dermal regeneration and decreased mechanical strength of dermal tissue was resulted from the cyto-toxic effect of SSD on dermal cells. Since the decreased mechanical strength may lead to reduction in resilience, toughness and maximum extension of the tissue, the identification of optimum dose for SSD that limits infection while minimizes the cyto-toxic effect may be clinically relevant.

• 대한한방내과학회지
• /
• 제35권4호
• /
• pp.416-427
• /
• 2014

Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

• Journal of Microbiology and Biotechnology
• /
• 제26권6호
• /
• pp.1140-1147
• /
• 2016

The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

• 생약학회지
• /
• 제46권1호
• /
• pp.59-64
• /
• 2015

Wound healing is a complex process that includes inflammation, granulation tissue formation, re-epithelialization, and remodeling. We reported previously that BuOH fraction from Hydnocarpi Semen (HS) crude extract exhibited wound healing activity in animal ulcer models. In this study, we investigated whether BuOH fraction activates keratinocyte and fibroblast via wound closure test and migration assay. In the scratch test, BuOH fraction accelerated the closure of a monolayer wound scratch at $100{\mu}g/mL$. After treatment with BuOH fraction for 18 h, keratinocytes showed a increase in migration at $25{\mu}g/mL$, whereas the migration of fibroblast increased significantly at $100{\mu}g/mL$ of BuOH fraction compared to control. The mechanism that the BuOH fraction of HS helps to promote healing by inflammation is possibly associated with the migration of keratinocyte and fibroblast.

• Archives of Pharmacal Research
• /
• 제28권11호
• /
• pp.1311-1316
• /
• 2005

To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.