• Archives of Pharmacal Research
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• 제29권7호
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• pp.556-562
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• 2006

Ten semi-synthetic derivatives of asiatic acid were prepared and their wound healing effects were evaluated by employing a tensile strength assay and a wound area assay. Among them, ethoxymethyl 2-oxo-3,23-isopropylidene-asiatate (12) showed the strongest and the fastest wound healing activity. Furthermore, it left the smallest scar after healing.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제50권4호
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• pp.206-215
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• 2024

Objectives: The current in vitro study aimed to assess the effects of ascorbic acid augmented albumin platelet-rich fibrin (AA Alb-PRF) on the wound healing activity of human gingival fibroblasts (HGFs) purported to be a regenerative biomaterial in surgical procedures. Materials and Methods: All assays were performed on three HGF groups, group I: complete media; group II: Alb-PRF, and group III: AA Alb-PRF. Alb-PRF was prepared following the protocol by Fujioka-Kobayashi et al. (2021). For preparation of AA Alb-PRF, 2,500 ㎍ AA was added to the blood pre-centrifugation. All groups were subjected to 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to estimate cell viability and proliferation, scratch assay for migration (0, 4, 12, and 24 hours) and transwell migration assay for chemotactic migration assessment (24 hours). Outcome variables were optical density (OD) for MTT assay, percentage of wound closure in scratch assay, and number of migrated cells in transwell migration assay. One-way ANOVA for MTT and transwell migration assays and two-way ANOVA for scratch assay with Bonferroni correction were performed with significance set at P<0.05. Results: Cell viability and proliferation (OD: 0.684±0.003 and proliferation: 28%) and wound closure (49.92%±1.62% at 4 hours and 61.39%±0.88% at 12 hours) were significantly higher in group III, while group II demonstrated the maximum number of HGFs migrating across the transwell membrane (9.25±2.49) with P<0.05. Conclusion: HGFs demonstrated a significant increase in viability and proliferation along with rapid wound closure in the presence AA Alb-PRF compared to Alb-PRF alone, indicating additional beneficial effects of AA. Thus, AA Alb-PRF potentiates the wound healing activity of HGFs and could be employed in oral, maxillofacial, and periodontal surgeries as a regenerative biomaterial.

• 대한임상검사과학회지
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• 제52권1호
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• pp.53-61
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• 2020

Cell migration은 embryogenesis 혹은 cancer metastasis 이외에, 물리적 손상에 의한 상처의 수복을 위해서 손상된 부위로 세포가 이동하는 매우 흔하게 관찰되는 현상 중 하나 이다. Wound healing assay는 in vitro의 이차원 평면상에서 세포의 이동을 관찰할 수 있는 기본적인 연구 기법이다. In vitro상에서 물리적 손상을 재현하는 가장 손쉬운 접근법으로서, 세포의 confluent monolayer 표면에 날카로운 도구를 이용하여 기계적인 스크레치를 내는 방법이 사용되고 있다. 완충 스프링이 탑재된 금속 핀을 96-well plate를 기반으로 하는 wound maker에 장착하여 multi-well plate 바닥 표면의 고르지 못한 굴곡과 스크레칭 팁 사이에 직각을 이루는 접촉면에서의 미세한 조절이 가능하도록 하였다. 실험용 팁으로 confluent monolayer위에 스크래치를 내었을 때에는 다양한 지그재그 패턴이 그려진 반면에, 직접 제작한 wound maker에서는 동일한 형태의 선형 wounds가 fibroblast가 seeding된 96-well plate의 각 well의 중심부에 그려짐을 확인하였다. 상용화 되어있는 몇몇 multi-well plate가 본 실험에서 제작된 wound maker와 호환되는 것을 고려하여 보았을 때에, 실시간 wound healing을 관찰하는 high content screening (HCS)실험에 있어서의 활용적인 측면에서 기존의 전형적인 polypropylene 파이펫 팁을 이용한 스크래칭 방법보다 더욱 용이한 방법임을 알 수 있다.

• Journal of Ginseng Research
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• 제47권1호
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• pp.133-143
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• 2023

Background: Past studies suggested that ginseng extracts and ginseng-derived molecules exerted significant regulatory effects on skin. However, no reports have described the effects of ginseng-derived nanoparticles (GDNPs) on skin cell proliferation and wound healing. In this study, we investigated whether GDNPs regulate the proliferation of skin cells and promote wound healing in a mouse model. Methods: GDNPs were separated and purified via differential centrifugation and sucrose/D2O gradient ultracentrifugation. GDNP uptake, cell proliferation and cell cycle progression were measured by confocal microscopy, CCK-8 assay and flow cytometry, respectively. Cell migration and angiogenic effects were assessed by the wound scratch assay and tube formation assay, respectively. ELISA was used to detect extracellular matrix secretion. The relevant signaling pathway was confirmed by western blotting. The effects of GDNPs on skin wound healing were assessed by wound observation, HE staining, and western blotting. Results: GDNPs possessed the essential features of exosomes, and they were accumulated by skin cells. Treatment with GDNPs notably enhanced the proliferation of HaCaT, BJ and HUVECs. GDNPs also enhanced the migration in HaCaT cells and HUVECs and angiogenesis in HUVECs. GDNPs increased the secretion of MMP-1, fibronectin-1, elastin-1, and COL1A1 in all three cell lines. GDNPs regulated cell proliferation through the ERK and AKT/ mTOR pathways. Furthermore, GDNPs facilitated skin wound healing and decreased inflammation in a mouse skin wound model. Conclusion: GDNPs can promote skin wound healing through the ERK and AKT/mTOR pathways. GDNPs thus represent an alternative treatment for chronic skin wounds.

• Nutrition Research and Practice
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• 제16권1호
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• pp.1-13
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• 2022

BACKGROUND/OBJECTIVES: Bitter taste receptors are taste signaling pathway mediators, and are also expressed and function in extra-gustatory organs. Skin aging affects the quality of life and may lead to medical issues. The purpose of this study was to better understand the anti-skin aging effects of bitter taste receptors in D-galactose (D-gal)-induced aged human keratinocytes, HaCaT cells. MATERIALS/METHODS: Expressions of bitter taste receptors in HaCaT cells and mouse skin tissues were examined by polymerase chain reaction assay. Bitter taste receptor was overexpressed in HaCaT cells, and D-gal was treated to induce aging. We examined the effects of bitter taste receptors on aging by using β-galactosidase assay, wound healing assay, and Western blot assay. RESULTS: TAS2R16 and TAS2R10 were expressed in HaCaT cells and were upregulated by D-gal treatment. TAS2R16 exerted protective effects against skin aging by regulating p53 and p21, antioxidant enzymes, the SIRT1/mechanistic target of rapamycin pathway, cell migration, and epithelial-mesenchymal transition markers. TAS2R10 was further examined to confirm a role of TAS2R16 in cellular senescence and wound healing in D-gal-induced aged HaCaT cells. CONCLUSIONS: Our results suggest a novel potential preventive role of these receptors on skin aging by regulating cellular senescence and wound healing in human keratinocyte, HaCaT.

• Archives of Plastic Surgery
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• 제37권1호
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• pp.7-11
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• 2010

Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.

• 생명과학회지
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• 제31권12호
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• pp.1100-1109
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• 2021

염생식물인 비쑥은 식용이 가능한 약용식물로서 살충, 항염, 항콜레스테롤, 해열, 항균 활성 등이 알려져 있다. 본 연구에서는 phorbol-12-myristate-13-acetate (PMA)로 자극된 인간 섬유육종 HT-1080 세포에서 5가지 활성검색방법 즉 : gelatin zymography, MMPs ELISA, wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot을 이용하여 비쑥의 조추출물과 그 용매 분획물의 MMP-2와 MMP-9 효소 활성에 대한 저해 효과를 평가하였다. 비쑥 시료들을 메틸렌 클로라이드(MC)와 메탄올(MeOH)로 각각 2번 추출하여 합한 것을 조추출물로 사용하였으며 이 조추출물은 MMP-2와 MMP-9 효소활성에 대해 유의한 억제 효과를 나타내었다. 이 조추출물은 용매극성에 따라 다시 n-hexane, 85% aq.MeOH, n-butanol 및 물 분획층으로 분획되었으며 이렇게 얻어진 4개의 용매 분획물들중에 n-hexane과 85% aq.MeOH 분획이 gelatin zymography와 MMP ELISA assay에서 MMP-2와 MMP-9의 활성을 효과적으로 억제하였다. 또한 wound healing assay, RT-PCR 및 Western blot assay에서 H₂O 분획을 제외한 모든 용매 분획물들이 세포 이동, 그리고 MMP-2 및 MMP-9의 mRNA와 단백질 발현을 유의하게 억제하였다.

• 한국응용과학기술학회지
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• 제33권4호
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• pp.788-794
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• 2016

Vinca alkaloids from plant Vinca minor have been investigated for their effects of tyrosinase inhibition, stimulation of ROS generation and increasement of cell migration activity. The methanolic crude extract and the water-soluble fraction exhibited $IC_{50}$ value of 3.1 mg/mL and 2.1 mg/mL. Vinca minor extract treatment significantly increased ROS levels in HaCaT cells, in a concentration-dependent manner. Treatments of Vinca minor extract led to increase wound closure when compared with non-treatment. Low dose (0.1% or 0.3%) of extracts have not significantly affected, compared with that in controls. By contrast, 0.5% extract have dramatic effect on wound healing activity of keratinocytes. Effects of Vinca minor extract in a filter-based cell mobility assay appear similar to that of wound closure assay, which suggests that the Vinca minor extract have wound healing effects on skin.

• 혜화의학회지
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• 제28권2호
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• pp.41-47
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• 2019

Objectives : Atopic dermatitis is an irritable skin disease accompanying rash and itching leading to impaired skin barrier. ATO-ALL is an ethanol extract of natural products comprising 12 herbs and effective on atopic dermatitis. In this study, we aimed to propose that the effect of ATO-ALL on skin regeneration in human keratinocyte cell line, HaCaT cells. Methods : To evaluate the skin regenerating effects of ATO-ALL, scratch wound healing assay, bromodeoxyuridine (BrdU) assay, and propidum iodide (PI) assay were performed using cultured HaCaT cell line. Result : Scratch wound healing assay showed that ATO-ALL was able to enhance the gap filling activity more than 2-fold at 7 ppm concentration compared with control group. BrdU assay demonstrated that ATO-ALL treatment increased the de novo cell proliferation in a dose-dependent manner. Finally, PI assay indicated that the cell cycle of HaCaT cells was modulated by ATO-ALL treatment. Conclusions : These results suggested that ATO-ALL may have skin regenerating effects by increasing cell proliferation via cell cycle regulation. Taken together, ATO-ALL is supposed to have a potential on regeneration of damaged skin or functional disease including atopic dermatitis.

• 환경생물
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• 제28권1호
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• pp.8-13
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• 2010

본 연구는 BRECs에서 AGEs로 유도된 세포의 이동 및 침윤에 있어서 AuNP의 역할에 관한 연구이다. 소 망막으로부터 내피세포를 분리하고, 세포 생존율은 MTT assay로 확인하였다. Wound migration assay는 BRECs의 이동력을 확인하기 위해 수행하였다. Tube formation은 on-gel system을 통해 확인하였다. AuNP의 apoptosis 유도는 caspase-3 assay를 통해 확인하였다. AGE-BSA은 세포증식 및 이동에 있어서 증가함을 보여주었다. 또한 AuNP는 AGE-BSA 존재 유무에 상관없이 세포의 증식, 이동, 신생혈관 형성을 억제하였고, caspase-3을 통해 apoptosis를 유도하였다. 이러한 결과, AuNP는 AGE로 유도된 신생혈관 형성 및 세포의 이동성을 억제하는 약재제로서, 당뇨성 합병증에 있어서 잠재적으로 중요한 분자가 될 것이다.