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

• The Korean Journal of Physiology and Pharmacology
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• 제21권2호
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• pp.189-195
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• 2017

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.

• Natural Product Sciences
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• 제25권1호
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• pp.38-43
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• 2019

Microbial wound infection prolonged the hospitalization and increase the cost for wound management. Silver is commonly used as antimicrobial wound dressing. However, it causes several adverse side effects. Hence, this study was aimed to evaluate the antimicrobial efficiency of Swietenia macrophylla seed extract on clinical wound pathogens. Besides, the bioactive constituents of the seed extract were also determined. S. macrophylla seeds were extracted with methanol by maceration method. The seed extract inhibited 5 test bacteria and 1 yeast on disc diffusion assay. The antibacterial activity was broad spectrum, as the extract inhibited both Gram positive and Gram negative bacteria. On kill curve analysis, the antibacterial activity of the seed extract was concentration-dependent, the increase of extract concentration resulted in more reduction of bacterial growth. The extract also caused 99.9% growth reduction of Bacillus subtilis relative to control. A total of 21 compounds were detected in gas chromatography- mass spectrometry analysis. The predominant compounds present in the extract were oleic acid (18.56%) and linoleic acid (17.72%). In conclusion, the methanolic extract of S. macrophylla seeds exhibited significant antimicrobial activity on clinical wound pathogens. Further investigations should be conducted to purify other bioactive compounds from the seeds of S. macrophylla.

• 대한한방내과학회지
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• 제35권4호
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• pp.416-427
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• 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.

• 대한예방한의학회지
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• 제18권1호
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• pp.83-92
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• 2014

Objective : This study was performed to investigate the antineoplastic effect of Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes and Trichosanthes kirilowii on SNU-80 anaplastic thyroid carcinoma cell line. Method : We examined whether our herbal medicines decreases cell growth rate of SNU-80 using MTT assay. We performed western blot analysis to verify that our herbal medicines induces apoptosis via caspase-dependent mechanism. We also performed wound healing assay and transwell invasion assay to investigate whether our herbal medicines affects the migration and invasion of anaplastic cancer cells, SNU-80. We also carried out ELISA assay to know our herbal medicines suppresses the expression of proinvasive molecules, such as VEGF and MMP-2 secreted from SNU-80. Results : MTT assay demonstrates that Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed strongly the growth of SNU-80. Western blot analysis demonstrates that Trichosanthes kirilowii induces apoptosis activating the cleavages of caspases (caspase-8, caspase-3) and PARP. Wound healing assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the migration of SNU-80. Transwell invasion assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the invasion of SNU-80. Elisa assay demonstrates that Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed the expression of VEGF and MMP-2. Conclusion : We could conclude that several herbal medicines suppresses the growth and inhibits the migration and invasion of SNU-80 which is anaplastic thyroid cancer cells. Especially, Rhus verniciflua Stokes, Trichosanthes kirilowii had stronger anti-cancer effect suggesting that we can apply them to treat anaplastic thyroid cancer.

• 한국식품저장유통학회지
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• 제14권4호
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• pp.419-424
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• 2007

하우스 포도 중 대표적인 것으로는 거봉, 델라웨어, 캠벨, 청포도를 열거할 수 있는데, 이에 대한 생물학적 활성 효과 중 항산화와 항암 활성에 대한 보고는 많으나 면역활성에 대한 활성의 연구는 미흡한 실정이다. 이에 우리는 4종의 포도의 메탄올 추출물로 항산화, 항암, 그리고 면역활성을 분석하였다. 전자공유능을 측정하기 위한 DPPH를 이용한 실험 결과 포도 4종에서 모두 항산화 활성을 나타내었으며, 환원력을 알아보기 위한 FRAP 실험 또한 활성을 나타내었다. 산화스트레스에 의한 세포사멸 억제 효과는 약간 있는 것으로 나타났다. 반면에 포도 추출물에 의한 cell proliferation 활성은 증가하는 것으로 나타났다. Nitric oxide (NO) 생성 억제 실험에서는 LPS에 의해 유도된 NO의 활성을 감소시키지는 않으나 포도 추출물 자제가 Raw 264.7 세포에서 NO의 활성을 유도하는 것으로 나타났다. Wound healing assay를 이용하여 항암효과를 알아본 결과 포도추출물 4종에서 세포의 운동성을 억제하는 효과를 가지는 것으로 나타났다. Mouse primary spleen cell 에서의 cytokine IL-4, IL-13의 활성을 알아본 결과 Con A로 유도된 IL-4와 IL-13의 발현 양을 현저히 줄이는 것으로 나타났다. 이로 미루어 보아, 포도 4종에 대한 생물학적 활성 결과 항산화, 항암활성은 물론 항천식 활성이 있는 것으로 확인되어 면역조절 활성이 포도의 기능성에 중요한 역할을 할 것으로 판단된다.

• 한방안이비인후피부과학회지
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• 제31권2호
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• pp.11-23
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• 2018

목적 : 본 연구에서는 $TNF-{\alpha}$와 $IFN-{\gamma}$로 자극한 인간피부각질형성세포 (HaCaT keratinocytes) 모델을 사용하여 지모가 피부장벽 기능에 미치는 영향을 알아보고자 하였다. 방법 : MTT assay를 통하여 지모 주정(70% 에탄올) 추출물 (EAA)이 HaCaT keratinocytes의 세포생존율에 미치는 영향을 확인하였으며 wound healing assay를 통해 EAA가 HaCaT 세포의 이주 능력에 영향을 주는지 관찰하였다. 또한 western blot analysis와 qRT-PCR을 통하여 EAA가 $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 발현 및 IL-4, IL-13, IL-6의 mRNA 발현, filaggrin의 단백질과 mRNA 발현에 미치는 영향을 조사하였다. 결과 : EAA는 처리 농도 $500{\mu}g/ml$까지 HaCaT keratinocytes의 세포생존율에 영향을 미치지 않았다. EAA는 wound healing assay에서 HaCaT 세포의 이주 능력을 증가시켰으며, $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 수준을 감소시켰다. 또한 EAA가 IL-4, IL-13, IL-6의 mRNA 발현을 억제하는 것 역시 확인할 수 있었다. 뿐만 아니라 EAA는 $TNF-{\alpha}/IFN-{\gamma}$ 자극에 의해 감소했던 filaggrin을 단백질과 mRNA 수준에서 회복시켰다. 결론 : EAA가 HaCaT 세포에서 Th2 type cytokines, pro-inflammatory cytokine의 억제와 filaggrin 회복을 통해 피부장벽 기능 손상에 대한 억제활성을 갖는 것을 확인하였으며, 이를 통해 EAA가 염증으로 인해 손상된 피부장벽 기능 개선에 효과적일 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5533-5537
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• 2013

MicroRNA-10b (miR-10b) has been reported to play an important role in some types of cancer, but the effects and possible mechanisms of action of miR-10b in the metastasis of nasopharyngeal carcinoma cells (NPC) have not been explored. The aim of the present study was to investigate the function of miR-10b in nasopharyngeal carcinoma and to determine the molecular mechanisms underlying its action. The MTT assay was used to assess proliferation of CNE-2Z cells. Wound healing and transwell migration assays were applied to assess cell migration and invasion, while and expression of E-cadherin and MMP-9 were detected using Western blot analysis. Real-time PCR was employed to detect the expression of genes related to migration and invasion and the $2^{-{\Delta}{\Delta}Ct}$ method was used to calculate the degree of expression. MTT assay showed the expression of miR-10b to have no effect on the proliferation of NPC cell lines. The wound healing assay showed that miR-10b mimics promoted the mobility and invasion of NPC cell lines. Inhibitors of miR-10b reduced the ability of NPC cell lines to migrate and invade. In addition, the expression of genes related to migration and invasion, such as E-cadherin, vimentin, and MMP-9, were confirmed to be different in the CNE-2Z NPC cell line transfected with miR-10b mimics and with miR-10b inhibitors. In the present study, miR-10b was found to upregulate the expression of MMP-9 and knockdown of miR-10b was found to significantly downregulate the expression of E-cadherin. On the whole, these results showed that miR-10b plays an important role in the invasion and metastasis of NPC cells.

• 대한한의정보학회지
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• 제15권1호
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• pp.57-66
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• 2009

Objectives : The aim of the present study is to evaluate the wound healing-enhancing and anti-inflammatory effects of Pinus densiflora, Cornus officinalis, Zingiber officinale, Ganoderma japonicum and Scutellaria baikalensison human keratinocyte, HaCaT cells. Methods : We adopted in vitro wound healing assay to measure the proliferation-and migration-enhancing effects in HaCaT cells. The expressions of cytokine genes were measured in HaCaT cells using real-time PCR analysis. Results : The extracts of Pinus densiflora, Cornus officinalis, Zingiber officinale, Ganoderma japonicum and Scutellaria baikalensis enhanced the proliferation and migration of HaCaTcells. The expression of keratinocyte growth factor receptor(FGFR2-IIIb) gene was also induced. The extracts inhibited iNOS, IL-$1{\beta}$ and TNF-$\alpha$ gene expression. Conclusions : The extract of Pinus densiflora, Cornus officinalis, Zingiber officinale, Ganoderma japonicum and Scutellaria baikalensis has wound healing-enhancing effects and anti-inflammatory effects.

• Nutrition Research and Practice
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• 제11권4호
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• pp.275-280
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• 2017

BACKGROUND/OBJECTIVES: Re-epithelialization has an important role in skin wound healing. Astaxanthin (ASX), a carotenoid found in crustaceans including shrimp, crab, and salmon, has been widely used for skin protection. Therefore, we investigated the effects of ASX on proliferation and migration of human skin keratinocyte cells and explored the mechanism associated with that migration. MATERIAL/METHOD: HaCaT keratinocyte cells were exposed to $0.25-1{\mu}g/mL$ of ASX. Proliferation of keratinocytes was analyzed by using MTT assays and flow cytometry. Keratinocyte migration was determined by using a scratch wound-healing assay. A mechanism for regulation of migration was explored via immunocytochemistry and western blot analysis. RESULTS: Our results suggest that ASX produces no significant toxicity in human keratinocyte cells. Cell-cycle analysis on ASX-treated keratinocytes demonstrated a significant increase in keratinocyte cell proliferation at the S phase. In addition, ASX increased keratinocyte motility across the wound space in a time-dependent manner. The mechanism by which ASX increased keratinocyte migration was associated with induction of filopodia and formation of lamellipodia, as well as with increased Cdc42 and Rac1 activation and decreased RhoA activation. CONCLUSIONS: ASX stimulates the migration of keratinocytes through Cdc42, Rac1 activation and RhoA inhibition. ASX has a positive role in the re-epithelialization of wounds. Our results may encourage further in vivo and clinical study into the development of ASX as a potential agent for wound repair.