• 한국축산식품학회지
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• 제36권2호
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• pp.237-243
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• 2016

Many researchers revealed that collagen contribute to maintaining the skin’s elasticity and inhibit wrinkling of skin. Korean native cattle (Hanwoo) bone (leg bone, foot and tail) infusion contains the various inorganic materials, collagen and chondroitin sulfate. All of this, a large quantity of collagen is included in Hanwoo infusion. Therefore, this study emphasized on the effects of collagen in the Hanwoo bone infusion. For the first time, Hanwoo bone infusions were directly added to the media of Human Dermal Fibroblast (NHDF-c) to test anti-aging effects. First, it was identified that growth rate of skin fibroblast was increased. Furthermore, the Hanwoo bone infusion increased a 50% of fibroblast collagen synthesis. Also, suppression of skin fibroblast aging was confirmed by treatment Hanwoo bone infusion. In conclusion, this study demonstrates the effects of infusion made from Hanwoo leg bone, foot and tail on anti-aging, wrinkle inhibiting and skin fibroblast elasticity maintaining. Therefore, this study identified that traditional infusion has effects that are good for skin elasticity.

• 대한의생명과학회지
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• 제13권1호
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• pp.55-60
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• 2007

Computerized image processing and analysis system was used for quantitative assessment of skin tissue components in color histological sections. The purpose of this study was to determine that the changes in the collagen fiber density and elastic fiber density in dermis in the rat skin as aging. And also to determine that the correlation between fibroblast density and collagen fiber density and elastic fiber density in the aged rat skin. Ten weeks old ($130{\sim}150g$) eight and fifty-four weeks old ($300{\sim}350g$) eight female Sprague-Dawley rats were used. The full-thickness skin biopsy specimens were prepared serial sections and stained with hematoxylin and eosin, Masson's trichrome and Verhoeff-van Gieson. The collagen fiber and the elastic fiber were identified using the image analysis processing system and then calculated the collagen fiber density rate and the elastic fiber density rate in the dermis. It also identified fibroblast and calculated fibroblast density in the dermis. By using a Student's t-test, a decrease in the collagen fiber density rate (t=-4.650, P<0.001) and the elastic fiber density rate (t=-6.494, P<0.001) of dermis can be observed in aged rats as compared with the young rats. A Student's t-test showed a significantly less fibroblast density in the aged rats than the young rats (t=-4.497, P<0.001). There were significantly positive correlation between the fibroblast density and the collagen fiber density rate (r=.69, P<0.001) and the elastic fiber density rate (r=.91, P<0.001). These results indicate that the aging may decrease the collagen fiber density and elastic fiber density due to reduced the proliferative and synthetic activity of fibroblast in the dermis.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.64-64
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• 2001

To facilitate the widespread application of somatic cell cloning, improvements in blastocyst production efficiency and subsequent fetal viability are required. Area where technical improvements are needed include donor cell treatments, starvation and passage numbers. This study was carried out to investigate the effect of serum-starvation and passage on the development of ear skin fibroblast cells cloned embryos. A skin biopsy was obtained from the ear of a 2-year-old Korean Hanwoo female. The cells were cultured in 10% FBS＋DMEM up to 2-3 months(up to 10 passages) and then used. In Experiment 1, the Korean bovine Ear Skin Fibroblast cells (KbESF) were either serum starved (culture in 0.05% FBS＋DMEM) or serum fed (10% FBS＋DMEM) for 4-7 days Prior to NT In Experiment 2, the KbESF cells used for nuclear transfer in these experiments were from passages 2 to 10. The development of 208 nuclear transfer (NT) embryos reconstructed from either serum starved or serum fed ear skin fibroblast was assessed. NT embryos reconstructed from serum starved and serum fed cells showed the same developmental rate (cleavage 80.16 vs. 85.37%; blastocyst 20.63 vs. 19,51%). The development of 590 nuclear transfer (NT) embryos reconstructed from passage 2 to 10 was assessed. We observed the same developmental rates for embryos derived from later Passages as compared with those embryos from early passages(blastocyst from 16.69 to 27.91%, average 20.17%). There was no significant difference between serum-fed and serum-starved donor cells. We observed no difference in developmental rates for embryos derived from 2 to 10 passages. These data show that prolonged culture and serum starvation does not affects the cloning competence of adult somatic cells.

• Radiation Oncology Journal
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• 제17권2호
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• pp.158-165
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• 1999

목적 : 유방암 환자의 피부 섬유모세포를 이용한 in vitro 배양 실험을 통하여 16 세포집락 비율 분포의 변화를 관찰하여 16 세포집락 비율과 in vitro 세포 노화 및 섬유모세포 공여자의 in vivo 연령과의 연관성을 조사하고자 하였다. 대상 및 방법 :유방암 수술을 받은 3명의 환자로부터 얻은 유방부위 피부를 본 실험대상으로 사용하였다. 각 환자의 유방부위 피부로부터 얻은 피부 섬유모세포 표본의 명칭을 C1, C2, C3a 및 C3b로 분류하였으며 각 표본 공여자의 연령은 C1이 44세, C2는 54세, 그리고 C3a 및 C3b는 동일한 공여자로서 연령은 55세였다. 피부 섬유모세포의 단일세포 부유액은 일차조직 배양법으로 얻었으며 100 개의 세포들을 100m1 의 조직배양 flask에 분주 후 37$^{\circ}C$에서 2주 동안 배양하였다. 5개의 flask에서 배양한 피부 섬유모세포의 16 세포집락 비율을 알기 위하여 crystal violet으로 염색한 후 10 배율의 입체현미경을 이용하여 16개 세포 이상으로 구성된 세포집락수를 1개 이상으로 구성된 세포 집락수로 나눈 수치를 16 세포집락 비율로 나타내었으며 각각 5개의 flask에서 얻어진 16 세포집락 비율 평균치를 각 계대배양에 대한 16 세포집락 비율로 나타내었다. C1, C2의 계대배양 횟수는 각각 12회, 17회 였으며 C3a와 C3b는 14회 계대배양 하였다. 결과 : C1, C2, C3a 및 C3b 피부섬유모세포 모두에서 16 세포집락 비율이 계대배양 횟수의 증가에 따라 감소하는 경향을 보였으며, 집단이배화증가에 따라 감소하였다. 그리고 계대배양 횟수가 증가함에 따라 집단이배화가 증가되는 것이 관찰되었으며 특히 C3a 섬유모세포의 상관계수가 0.954(P=0.0001)로서 가장 강한 상관관계가 있음을 보였다 동일한 지점의 집단이배화에서 16 세포집락 비율이 고연령자인 C3a 공여자보다 저 연령자인 C1 공여자에서 더 높게 나타났다. 결론 : 사람 피부 섬유모세포의 in vitro 배양에서 계대배양 횟수의 증가에 따라 집단이배화는 증가되고, 세포 노화로 인해 16 세포집락 비율은 감소되는 것을 알 수 있었다. 또한 저연령의 피부 섬유모세포일수록 집단이배화 증가에 따른 16 세포집락 비율 감소가 고연령의 경우보다 완만하였다. 따라서 피부 섬유모세포 in vitro 배양에서 관찰되는 16 세포집락 비율은 in vitro 세포노화의 지표로서 유용하며 또한 피부 섬유모세포 공여자의 연령 평가에 이용될 수 있을 것으로 사료된다.

• 한국응용과학기술학회지
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• 제38권5호
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• pp.1249-1254
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• 2021

UV는 산화 스트레스를 유발하고 MMP(Matrix Metalloproteinase) 발현을 증가시켜 피부 노화를 발생시킨다. 따라서 자외선로 인한 피부 손상을 예방하면 피부 노화를 감소시킬 수 있다. 스피룰리나는 강력한 항산화제로 원핵생물로 구성되어 있다. 본 연구는 피부 섬유아세포를 사용하여 UVB 방사선에 대한 스피룰리나 유래 피코시아닌(PC)의 광보호 효과를 조사했다. 그 결과, PC는 섬유아세포 생존율 측면에서 5-40 ㎍/mL 농도에서 독성을 나타내지 않았다. UVB 조사된 섬유아세포의 생존율은 50.5%였으며 PC 처리로 73.5%로 증가했다. MMP-1 및 MMP-9 발현은 UVB 처리로 증가하는 반면 PC 처리한 군에서 감소했다. 이러한 결과를 종합해보면 PC는 UVB 조사로 인한 산화적 손상과 피부노화와 관련된 인자를 감소시켜 노화를 예방할 수 있을 것으로 사료된다.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.13-34
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• 2003

Fructose-1, 6-diphosphate (FOP), a glycolytic metabolite is reported to ameliorate inflammation and inhibit the nitric oxide production in murine macrophages stimulated with endotoxin. It is also reported that FOP has cytoprotective effects against hypoxia or ischemia/reperfusion injury in brain and heart. In this study, we examined whether FDP has protective effects on UV-induced oxidative damage in skin cell culture system and human skin in vivo. FDP had a protective role in UVB-induced LDH release and ROS accumulation in HaCaT although it did not show direct radical scavenging effect in the experiment using 1, 1-diphenyl-2-picrylhydrazyl (DPPH). FDP also preserved cellular GSH content after UV irradiation in HaCaT and normal human fibroblast culture system. Cellular oxidative stress induces multiple downstream signaling pathways that regulate expression of multiple gene including MMP-1 and collagen, we examined the effects of FDP on UV-induced alteration of these protein expression in fibroblast culture and human skin in vivo. The increased MMP-1 expression in fibroblast and human skin by UV irradiation was significantly decreased by FDP. FDP also prevented the UV-induced decrease of collagen expression in fibroblast and human skin. Moreover, the decreasing the intracellular levels of reducing equivalents in human fibroblast by glutathione (GSH) depletion lowered the UVA dose threshold for reduction of procollagen expression, indicating that the differences of glutathione contents define the susceptibility of fibroblasts towards UV-induced reduction of procollagen expression. FDP also preserved cellular GSH content after UV irradiation, indicating that FDP has protective effects on UV-induced reduction of procollagen expression, which are possibly through maintaining intracellular reducing equivalent. Based on these premises, we examined the effect of daily use of a moisturizer containing FDP on facial wrinkle in comparison with vehicle moisturizer lacking FDP. In the clinical study, FDP significantly decreased facial wrinkle compared with vehicle alone after 6 months of use. Our results suggest that FDP has anti-aging effects in skin by increasing cellular antioxidant system and preventing oxidative signal and inflammatory reaction. Therefore FDP may be useful anti-aging agent for cosmetic purpose.

• Animal cells and systems
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• 제10권1호
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• pp.15-20
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• 2006

Allergic inflammation is thought to be a Th2 cell-dominant immune response during which tissue-resident fibroblasts produce chemokines which contribute to the recruitment of migratory leukocytes to sites of tissue injury. Thymus and activation-regulated chemokine (TARC; CCL17) is a potent member of the CC chemokine family and a selective chemoattractant for Th2 cells. In order to study the regulatory profiles of TARC production by $TNF-{\alpha}$, $IFN-{\gamma}$, and Il-4 in human normal skin fibroblast, CCD-986sk cell line was used. The expression of TARC protein was measured using ELISA, and mRNA level was detected by RT-PCR. The combination of $TNF-{\alpha}$ and IL-4 induced a time-and dose-dependent synergistic increase in the expression of TARC at both protein and mRNA levels in the cultured human skin fibroblasts. Exposure of the cells to single cytokine had no effect on TARC expression. The high concentration (100 ng/ml) and long incubation time (72 h) of $IFN-{\gamma}$ further enhanced the TARC production induced by $TNF-{\alpha}$/lL-4 in the skin fibroblast. This synergistic effect of Th1 and Th2 type cytokines on TARC production by skin fibroblasts may contribute to the inflammatory cell infiltration and tissue damage with allergic inflammation.

• 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.

• 생약학회지
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• 제25권3호
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• pp.249-257
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• 1994

The cytotoxic and antitumor activity of Perilla frutescens extract on cultured 3T3 fibroblast and skin melanoma cells were evaluated by tetrazolium MTT (MTT) and neutral red (NR) colorimetric assay methods. Lactate dehydrogenase activity was also measured. The light microscopic study was carried out to observe morphological changes of cultured mouse fibroblast and skin melanoma cells. The results were as follows: 1. Water and ether extracts showed a significant cytotoxicity in 3T3 fibroblast and all extracts exhibited a significant anti-tumor activity in skin melanoma cells. Methanol, ethyl acetate and ethanol extracts showed low cytotoxic effects, but exhibited a high anti-tumor activity. 2. The MTT absorbance in 3T3 fibroblast was significantly decreased by treatment with ether, water, chloroform and ethanol extracts and skin melanoma cells was significantly decreased by treatment with all extracts. The difference in MTT absorbance in two cell types was most remarkable when treated with methanol and ethanol extracts. 3. Methanol and ethyl acetate extracts showed the strongest effect in growth inhibition of melanoma cells. These results indicated that methanol extract possessed a low cytotoxicity and a strong anti-tumor activity.

• KSBB Journal
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• 제29권1호
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• pp.42-49
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• 2014

The aim of this study was to develop a composite human skin equivalent for wound healing. Collagen type1 and acellular dermal matrix powder were utilized as the scaffold with dermal fibroblasts and keratinocytes for the development of a composite human skin equivalent. Fibroblast maintained the volume of composite skin equivalent and also induced keratinocytes to attach and proliferate on the surface of composite skin equivalent. The composite human skin equivalent had a structure and curvature similar to those of real skin. Balb-C nu/nu mice were used for the evaluation of full-thickness wound healing effect of the composite human skin equivalent. Graft of composite skin equivalent on full-thickness wound promoted re-epithelialization and granulation tissue formation at 9 days. Given the average wound-healing time (14 days), the wound in the developed composite skin equivalent healed quickly. The overall results indicated that this three-dimensional composite human skin equivalent can be used to effectively enhance wound healing.