• 대한한방부인과학회지
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• 제25권1호
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• pp.20-33
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• 2012

Objectives: This study was performed to elucidate the effects of Kanghwalsokdan-tang extract(KS) on hyper-plasy of collagen and cell damage in UVB-irradiated dermal fibroblast. Methods: To demonstrate the effects of KS on wound healing we used human dermal fibroblast(F6). We evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit. Also, we measured the nitrite production, and LDH release in UVB-irradiated dermal fibroblast to elucidate the action-mechanism of KS. Results: 1. KS decreased the cell proliferation of dermal fibroblast. 2. KS decreased the biosynthesis of collagen in dermal fibroblast. 3. KS decreased the synthesis of TIMP-1 in dermal fibroblast. 4. KS had no effect on the LDH-release of UVB-irradiated dermal fibroblast. 5. KS inhibited nitrite production in UVB-irradiated dermal fibroblast. Conclusions: From the results, we concluded that KS has a protective effect on wound healing and photoaging.

• 대한한방부인과학회지
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• 제24권2호
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• pp.13-32
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• 2011

Objectives: This study was performed to elucidate the effects of Danchisoyo-san (DS) on cell damage and gene expression in UVB-exposed dermal fibroblast. Methods: To demonstrate the inhibitory effects of DS on aging of the skin, we used human dermal fibroblast(F6) and UVB light(30 mJ/$cm^2$) was used to damage to dermal fibroblast. We measured the nitrite production, LDH release, and gene expression in UVB-irradiated dermal fibroblast to elucidate the actionmechanism of DS. Also, we evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit, and gene expression (MMP-1, procollagen, c-fos, c-jun, NF-kB, Bcl-2, Bcl-xL, iNOS) were determined using real-time PCR. Results: 1. DS inhibited LDH-release, nitrite production in UVB-irradiated dermal fibroblast. 2. DS suppressed the gene expression of MMP-1 in UVB-irradiated dermal fibroblast. 3. DS increased the gene expression of procollagen in UVB-iradiated dermal fibroblast. 4. DS suppressed the gene expression of c-jun, c-fos, NF-kB, iNOS in UVBirradiated dermal fibroblast. 5. DS increased the gene expression of Bcl-2 in UVB-iradiated dermal fibroblast. 6. DS increased the cell proliferation of dermal fibroblast. Conclusions: From the results, we concluded DS increases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that DS has the antiwrinkle effects.

• 대한한방부인과학회지
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• 제24권4호
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• pp.10-19
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• 2011

Objectives: This study was performed to elucidate the effects of Kwibi-tang extract(KB) on dermal fibroblast. Methods: To demonstrate the effects of KB on dermal fibroblast, we used human dermal fibroblast(F6) and UVB light(30 $mJ/cm^2$) was used to damage to dermal fibroblast. we measured the nitrite production, LDH release in UVB-irradiated dermal fibroblast to elucidate the action-mechanism of KB. Also, we evaluated cell proliferation of dermal fibroblast and the amount of increased PICP, TIMP-1 in dermal fibroblast. Results: 1. KB decreased the cell proliferation of F6 dermal fibroblast in concentration of 50 ${\mu}g/ml$. 2. KB decreased the synthesis of PICP in concentration of 50 ${\mu}g/ml$. 3. KB decreased the synthesis of TIMP-1 in concentration of 50 ${\mu}g/ml$. 4. KB have no effect on the damage in UVB-irradiated F6 dermal fibroblast. Conclusions: From the results, we concluded KB decreases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that KB has the anti-hyperplasy of dermal fibroblast.

• Archives of Plastic Surgery
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• 제34권4호
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• pp.420-425
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• 2007

Purpose:The mechanism of scar formation is not fully understood. Fibroblast is an important cell in wound healing process. We experienced a patient who was taking progesterone orally. Upper blepharoplasty was performed on her but, wound healing was delayed. We hypothesized that progesterone was the cause of delayed wound healing and fibroblast proliferation inhibition. We investigated the effect of progesterone in vitro on human dermal fibroblasts to study the effects on fibroblast proliferation. Methods: Human dermal fibroblasts from four persons were cultured initially. Progesterone is mixed to them at various concentrations, and fibroblast cell count was measured by MTT assay method at 570 nm. We confirmed that progesterone has some inhibitory effect on fibroblast proliferation and maximal inhibitory concentration of progesterone was determined. Then fibroblasts from a total of nineteen persons were cultured and the effects of progesterone were studied. Results: The initial study showed the maximal inhibitory concentration of progesterone to be $50{\mu}g/ml$. The main study showed that progesterone had 70.9% inhibitory effect on human dermal fibroblast in vitro. Conclusion: Progesterone has inhibitory effect on cultured human dermal fibroblast proliferation in vitro.

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

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

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권3호
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• pp.211-220
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• 2007

Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

• 약학회지
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• 제51권4호
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• pp.229-234
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• 2007

We investigated antioxidative activity of the water and ethanol extracts of leaves of Acanthopanax divaricatus var. albeofructus in human dermal fibroblast (HDFs) irradiated by UVA. The irradiation of UVA did not affect the cell viability of HDFs. The antioxidative activity of the extract was investigated by xylenol orange, TBARS (thiobarbituric acid reactive substances) and antioxidant enzyme assay. Both extracts showed H202 scavenging activity and inhibited lipid peroxidation in HDF cells irradiated by UVA. The extracts also recovered enzyme activity in the same cells.

• Molecular & Cellular Toxicology
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• 제4권2호
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• pp.150-156
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• 2008

The photoprotective effects of minerals from Korean indigenous ores, consisting mainly of sericite, on UVA-irradiated human dermal fibroblast (HDF) were examined. Zymographic analysis showed that the treatment of the minerals significantly reduced the UVA-enhanced MMP-1 activity and mRNA level. The minerals also showed strong inhibitory effect on MMP-2 activity and mRNA expression. Moreover, the minerals were better than polyphenol in reducing MMP-1 and MMP-2 expressions. Notably, the minerals significantly enhanced collagen biosynthesis in the HDF. Inhibition of the elastase activity and protection against the oxidatively damaged HDF cell were also found in the presence of the minerals. Taken together, the ore minerals may be used as the potent photo-protective and anti-skin-aging ingredients which can prevent skin cell damage by UVA.

• 대한화장품학회지
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• 제39권3호
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• pp.177-186
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• 2013

피부 섬유아세포는 인간 피부의 주요 콜라겐 생산 세포이다. 노화가 진행되면, 섬유아세포에서의 콜라겐 생산이 감소되고, matrix metalloproteinase-1 (MMP-1)에 의해 시작되는 콜라겐 조각화가 증가된다. 즉 섬유아세포의 콜라겐 항상성의 불균형으로 인해 피부 collagenous, 세포외기질(ECM)의 구조와 기능이 변형되어, 피부노화가 촉진되는 것이다. Cysteine rich protein 61 (CCN1)는 CCN family의 일부이며, 인간피부의 섬유아세포에서 콜라겐 항상성을 조절하는 단백질이다. 노화된 인간 피부 섬유아세포에서의 CCN1 과 발현은 실질적으로 유형 I procollagen 생성을 감소시킴과 동시에 MMP-1의 발현을 증가시켜 섬유의 콜라겐 저하를 일으킨다. 그리고 노화된 섬유아세포는 노화 전 섬유아세포에 비해 증식률이 감소한다. 본 연구에서 만들어 사용한 복제 노화 피부 섬유아세포는 유형 I procollagen의 생성량이 감소하였고, MMP-1의 발현 수준이 증가하는 특징을 나타냈다. 또한 CCN1 단백질의 발현이 증가되고, 증식률이 감소하는 특징을 나타냈다. 가수분해 구멍갈파래 추출물은 노화 전 섬유아세포에서 새로운 콜라겐의 합성을 촉진하고 자외선에 의해 증가된 MP-1의 발현을 감소시켜 광노화를 개선하는 물질로 알려져 있다. 본 연구에서는 이러한 활성을 나타내는 가수분해 구멍갈파래 추출물을 사용하여, 복제 노화 피부 섬유아세포에서 가수분해 구멍갈파래 추출물에 의한 CN1 단백질의 발현 억제 여부를 조사하였으며, 이들 추출물은 배양된 복제 노화 피부 섬유아세포에서 유형 I procollagen의 생성을 증가시켰으며, MMP-1 발현을 억제시키는 것을 확인하였다. 또한, 콜라겐 항상성을 조절하는 단백질인 CN1 발현을 크게 감소시켰으며, 노화세포의 증식률을 증가시켰다. 이 결과는 복제 노화 섬유아세포가 in vitro 자연 노화모델로 화장품 원료 활성 연구에 사용될 수 있음을 말한다. 그리고 가수분해 구멍갈파래 추출물은 광노화 뿐 아니라 자연노화를 개선하는 피부미용제로 주름개선 기능성 화장품에 사용가능 하다는 것을 의미한다.