• Korean Journal of Acupuncture
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• 제37권2호
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.192-202
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• 2003

The wrinkles correspond to the most obvious expression of skin ageing and are manifested by changes on the organization and dermal structure. In the extracellular matrix, decreased quantities of collagens and glycosaminoglycans as well as a deterioration of the fibrillary network is noted, result in a reduction of dermal thickness. In addition, the activity of the collagenases increases in contrast to the synthesis of collagen fibers. Nor are cells spared during the aging process. We thus studied and compared the contractile capacity as well as the synthesis capacity of normal human fibroblasts and human fibroblasts obtained from biopsies of forehead wrinkles. The capacity of the fibroblasts to be adhered to the collagen network and to maintain a three-dimensional structure of dermis was studied on a model of equivalent dermis. The metabolic activity was studied by evaluating the capacities of synthesis of collagen I, main component of dermis. Human fibroblasts resulting from the forehead wrinkle contract less the gel of collagen than the normal human fibroblasts and present an activity of biosynthesis of collagen I less important than normal human fibroblasts. These results show that fibroblasts with aging present a deceleration of their metabolic activity and lose their capacity of adhesion to collagen fibers thus limiting the possibility of organizing the dermal tissue. We investigated the potential of an active ingredient able to compensate for the reduction of the metabolic activity and to restore the contractile capacity of fibroblasts obtained from forehead wrinkles. This effect was compared with a reference molecule: the vitamin C.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.799-821
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• 1996

The ultimate goal of periodontal treatment has been to facilitate regeneration of diseased periodontal tissues, destroyed by inflammatory periodontal disease. For regeneration of the periodontium to occur, all of component tissues must be restored to their original position and architecture. Growth factors which were known to promote the cellular processes, ie, proliferation, migration and matrix synthesis, have been in the spotlight of current periodontics. Platelet-derived growth factor(PDGF) stimulates collagen and non collagen protein synthesis, migration and proliferation of periodontal ligament cells. Insulin-like growth factor(IGF) has potentials to induce collagen and bone matrix synthesis so that it regulates normal bone remodeling. Application of the combination have been known to facilitate formation of bone and cementum, and to synergistically interact to promote coronal migration and proliferation of periodontal ligament cells. These two growth factors have been reported to exhibit positive effect in the periodontally diseased teeth or class m furcation defects. The aim of the present study was to test the hypothesis that PDGF-BB alone or the combination of PDGF-BB and IGF-I can predictably enhance regeneration of the periodontium in the dehiscence defect. Following the resection of premolars, roots were embedded. After 12 weeks of healing period, standardized experimental $4{\times}4mm$ dehiscence defects were created on the mid-facial of the premolar roots in each of 4 young adult dogs. In control group, only methylcellulose gel was inserted in the defects. In experimental group I and II, gel with $2{\mu}g$ of PDGF-BB or $2{\mu}g$ of PDGF-BB and $1{\mu}g$ of IGF-I was inserted in the defects, respectively. At 8 weeks postsurgery, the dogs were sacrificed. The results were observed histologically and analyzed histomorphometrically.The results of this study were as follws. 1. The new cementum formation was $1.26{\pm}0.69mm$ in the control group, $1.80{\pm}0.84mm$ in the experimental group I, $1.93{\pm}0.51mm$ in the experimental group II. The experimental group III, the experimental group I, the control group were in the order of cementum formation without statistically significant differences between control and all experimental groups. 2. The new bone formation was $1.00{\pm}0.53mm$ in the control group, $1.53{\pm}0.63mm$ in the experimental group I, $l.33{\pm}0.45mm$ in the experimental group II. The experimental group I, the experimental group II, the control group were in the order of bone formation without statistically significant differences between control and all experimental groups. 3. The root resorption was $1.12{\pm}0.64mm$ in the control group, $1.34{\pm}0.73mm$ in the experimental group I, $0.79{\pm}0.59mm$ in the experimental group II without statistically significant differences between control and all experimental groups. These results suggested that the use of PDGF-BB alone or PDGF-BB and IGF-I in the dehiscence defects might facilitate periodontal regeneration in some degree, but has not shown statistically significant results.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.99-105
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• 2000

Cell-Cell interaction and the extracellular matrix (ECM) are belisved to play essential roles during in vitro culturing of primary hepatocytes in the control of differentiation and in the maintenance of tissue spcific functions. The objective of this study was to examine the effects of degree of cell-cell contact (DCC) on liver sperific function of rat promary hepatocytes. Hepatocyte aggregates with various with various degrees of cell-cell contantact, I. e., dispersed cell, longish aggregate, rugged aggregate, and smooth spheroid were obtained at 1, 5-6, 15-20, and 36-48 hrs, respectively in suspension cultures grown in spinner flasks embedded in Caalginate bead and collagen gel in order. The may result from mass transfer limitation and shear damage caused by agitation during aggregation. The rugged aggregate showed a higer viability and albumin secretion rate than the dispersed cells or the other aggregates. This result indicates the possible enhancement of a bioartificial liver's (BAL) performance using primary hepatocytes and the reduction in time to prepare a BAL through optimization of the immobilization time.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.449-452
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• 2001

In this study. an efficacy study of Portulaca Extract (PE) and ${\beta}$ -glucan. candidates for cosmetic additives. was pedormed using artificial skin model (AS). The AS consists of collagen gel matrix populated by ATCC human skin fibroblast cell line that is overlaid with epidermal human skin keratinocyte cell line. Cytotoxicity and anti - inflammatory activity of PE and ${\beta}$ -glucan were assessed using monolayer and AS models by measuring cell viability and the secretion of interleukin -1 ${\alpha}$.

• Journal of Periodontal and Implant Science
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• 제41권5호
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• pp.227-233
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• 2011

Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.

• 한국임상수의학회지
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• 제32권3호
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• pp.224-230
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• 2015

본 연구는 키토산 알지네이트 수화겔을 사용하여 제작된 연골세포의 3차원 구조를 유지하며 생물학적, 생리학적인 기능을 유지하는데 적합한 poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) 지지체의 효과에 대한 연구이다. 체내에서 수화겔은 단독으로 지지체 역할을 하기에는 부하를 견디기에 약하다. 이에 본 연구에서는 연골세포와 유사한 세포, 세포외 기질의 3차원적 구성을 만들기 위해 PLCL 지지체와 수화겔을 사용하여 합성 지지체를 제작하였다. 염화나트륨을 사용한 입자 침출 기법으로 85%의 다공성, $300-500{\mu}m$ 크기의 구멍을 가진 탄성력 높은 지지체를 제작하였다. 소의 연골세포와 키토산 알지네이트 겔 혼합물이 PLCL 지지체에 적용되었고 대조군의 알지네이트와 비교 연구하였다. 키토산 알지네이트 수화겔과 연골세포가 혼합된 경우에 알지네이트 단독 사용에 비해 세포 성숙, 증식, 세포외 기질의 합성, sGAG 생성과 II 형 콜라겐의 발현 등의 효과가 좋은 것으로 확인되었다. 본 연구 결과를 통해 PLCL 지지체에 연골세포와 키토산 알지네이트 겔 혼합물을 적용할 경우 세포 증식과 기질의 합성에 적합한 환경을 만들 수 있으며 연골의 복구와 재생에 효과적으로 사용될 수 있을 것으로 기대된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제19권3호
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• pp.265-286
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• 1997

Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

• 대한화장품학회지
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• 제30권2호
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• pp.241-246
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• 2004

발아 과정을 통한 식물 종자의 변화를 확인하고, 발아된 식물종자로부터 새로운 펩타이드를 개발하여 항노화 화장품 소재로서의 가능성을 확인하였다. 발아과정을 통한 식물 종자의 변화는 단백질 양 측정, 겔 투과 크로마토그램(GPC)과 SDS-PAGE를 통한 단백질 분자량 분포 변화, 아미노산 조성분석을 통해 발아 전과 비교하였다. 발아된 검은 쌀로부터 펩타이드를 생산하기 위해 단백질 분해효소를 처리하였으며, 생산된 발아 검은 쌀 펩타이드의 효과를 발아 검은 쌀 단백질과 비교하여 확인하였다. In vitro matrix-metalloprotease (MMP) 활성 저해효과는 형광 정량법을 이용하였으며, 인간 섬유아세포 활성화 효과를 확인하였고, 콜라겐 생합성 촉진효과와 자외선 조사에 의한 MMP 발현 저해효과는 효소면역분석법(ELISA)을 이용하여 확인하였다. 발아에 의해 검은 쌀의 단백질량, 단백질 분자량 분포 및 아미노산의 조성이 변화함을 확인하였으며, 검은 쌀의 발아와 단백질 분해효소중 파파인 처리를 병행함으로서, 상대적으로 낮은 분자량을 갖는 신규 펩타이드의 제조가 가능하였다. GPC를 이용하여 제조된 발아 검은 쌀 펩타이드의 분자량 분포를 확인한 결과, 대부분이 900dalton 이하의 분자량을 가지는 것을 확인하였으며, 또한, 이러한 발아 검은 쌀 펩다이드는 약 40% 정도의 인간 섬유아세포 활성화 효과를 가지며, 또한 농도 의존적으로 콜라겐 분해효소 활성 저해효과를 가지는 것을 확인하였다. 또한, 자외선 조사에 의해 유도되는 콜라겐 분해효소 발현을 약 50% 억제하며, 콜라겐 생합성을 약 20% 촉진하는 것을 확인할 수 있었다.

• 대한약리학회지
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• 제26권2호
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$는 많은 세포의 증식, 분화 및 여러가지 세포기능에 대해 다양한 조절기능을 갖고 있는 multifunctional polypeptide로 알려져 있다. $TGF-{\beta}$는 골기질에 상당량 존재하며 골조직 대사에 대해 광범위한 영향을 나타낸다. 여러가지 연구결과에 의해 기질과 연관된 $TGF-{\beta}$는 골세포 자체의 산물로 여겨지고 있으나 골조직세포군중 어느종류의 세포가 $TGF-{\beta}$를 형성하는지에 대해서는 논란이 되고 있다. 본 연구는 $TGF-{\beta}$를 형성하는 골조직세포를 규명하고 $TGF-{\beta}$가 서로 다른 세포들에 미치는 영향을 관찰하고자 하였다. 본 실험에 필요한 특정골조직세포군을 얻기 위하여 백서태자두개관을 연속효소처리하여 수집한 골조직세포군의 생화학적 특성규명을 시행하였다. 효소 처리후 초기에 유리되는 세포는 섬유아세포의 특성을 보이며 후기에 유리되는 세포는 acid 및 alkaline phosphatase 활성과 부갑상선호르몬, calcitonin, prostaglandin $E_{2}$에 대한 c-AMP의 반응 및 교원 단백질합성등을 통해 볼때 조골세포유사세포로 보여진다. 골조직과 두개관세포 추출물의 Polyacrylamlde gel과 immunoblot analysis에 의해 골조직내에 $TGF-{\beta}$의 존재와 골세포에 의한 $TGF-{\beta}$의 생성을 확인하였다. 두개관 세포 추출물의 분석에서 모든 세포군에서 $TGF-{\beta}$가 합성되는 것을 관찰하였다. 외부에서 $TGF-{\beta}$를 가한 경우 무혈청 배지에서 골조직 증식에 대해 두가지 양상의 반응을 보였다. 조골세포 유사세포에서는 촉진하는 반응을 보였으나 섬유아세포군에서는 억제반응을 나타냈다. 반면에 교원 및 비교원 단백질 합성에 있어서는 모든 두개관세포군이 촉진반응을 보였다. 단백질 합성증가는 교원단백에 특이성이라기 보다는 일반적인 증가로 보여진다. 또한 단백질합성증가에 대한 $TGF-{\beta}$의 영향은 세포증식과의 관련성이 없는 것으로 사료된다. 결국 골세포에 의한 $TGF-{\beta}$의 생성과 여러 세포군에 대한 서로 다른 작용으로 보아 $TGF-{\beta}$는 autocrine과 paracrine 양상으로 세포기능을 조절함으로써 골조직 대사를 조절하는 중요한 기능을 발휘하는 것으로 사료된다.