• KSBB Journal
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• 제24권3호
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• pp.312-320
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• 2009

치료용 단백질을 생산하는 생물의약품 산업이나 세포치료제 및 이식용 세포를 다루는 재생의학 분야 등의 세포기반 산업에서 안정적인 세포의 보존은 필수적인 요소이다. 본 연구에서는 인간 피부 섬 유아세포의 $4^{\circ}C$ 저온보존에서 우수한 성능을 나타내는 개선된 저온보존액을 개발하고, 저온에 의한 세포 손상을 보호함으로써 보다 안정적인 세포 저온보존 기술을 제공하고자 하였다. 세포의 저온보존에서 우수한 효능을 나타내는 핵심 성분을 탐색한 결과, yeast hydrolysate 등의 단백질 가수분해물을 첨가한 보존액에서 월등히 뛰어난 보존효과가 나타남을 확인하였다. 단백질 가수분해 물은 미생물, 식물, 동물유래 단백질 가수분해 물에서 모두 우수한 효과를 나타냈으며, 특히 단백질 가수분해물 성분 중 분자량 10kDa 이하의 펩타이드를 첨가한 저온보존에서 우수한 보존효과가 나타났다. 저온에 의한 세포손상에 대해 단백질 가수분해물은 세포내 ATP level의 감소를 막아주고 ROS 생성을 억제하는 것으로 나타났으며, 항산화제 및 삼투압 조절물질을 단백질 가수분해 물과 함께 첨가하였을 때 더욱 우수한 세포 보존효과를 보였다. 최종적으로 본 연구에서 개발한 KUL261 저은보존액 (DMEM/F12 1 : 1 medium, yeastolate 1%, $\alpha$-tocopherol $100{\mu}M$, dextran 2.5%)은 기존의 저온 보존액에 비해 세포 생존을 및 성장률에서 월등히 우수한 성능을 나타내었다. 결론적으로, 핵심 유효성분으로 단백질 가수분해물을 포함하는 개선된 저온보존액은 기존의 보존액보다 월등히 우수한 보존효과를 제공하며, 세포치료제 및 재생의학 분야의 발전과 글로벌 상업화에 기여할 수 있을 것이다.

• Journal of Oral Medicine and Pain
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• 제38권2호
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• pp.215-219
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• 2013

하행성 억제계란 중뇌, 연수, 뇌교에 존재하는 해부학적 유해수용 조절성 기전을 일컫는 용어이다. 이들 부위를 전기적으로 자극을 하면 진통효과가 나타나며, 하행성 억제계의 실패시 지속적인 통증이 야기된다는 것을 알 수 있다. 또한 우울불안 같은 질환은 만성 신경병성 통증 상태로 쉽게 진행됨이 밝혀졌다. 이러한 요인들이 만성 신경병성 통증에 영향을 주는 경로는 아마도 하행성 억제계일 가능성이 있다. 흥미롭게도, 광범위하게 하행성 억제계가 작동하지 않을 경우 과민성 대장증상이 호발하는 것으로 보인다. 또한 이러한 환자들은 높은 불안, 우울 지수가 관찰되기도 한다. 다양한 연구에서, 하행성 억제계에 관여하는 ${\alpha}2$ 아드레날린성 약물, 아편유사약물들이 만성 통증에 사용될 수 있음을 동물에서 평가 중이다. 아직 신체내에서 얼마나 하행성 억제계가 일어나고 있는가에 대해서는 임상적으로 증명하기 힘든 감이 있지만, 여러 감각 신경기전의 수정에 중요한 구실을 하고 있는 것으로 믿어진다. 즉 중추신경계는 대상을 인식하기 위해 말초정보를 받아들이는 기능만 있는 것이 아니라 여러 방법으로 정보의 홍수를 조절하고 선택하는 기능을 동시에 갖추고 있는 것으로 생각된다.

• Journal of Periodontal and Implant Science
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• 제24권1호
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• pp.144-154
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• 1994

Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

• 구강회복응용과학지
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• 제28권2호
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• pp.201-212
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• 2012

임플란트 주위조직과 치아 주위조직은 조직학, 형태학적으로 유사한 면이 있다. 특히 functional ankylosis라 불리는 티타늄과 연조직의 직접적인 상피부착은 치아주위 조직의 접합상피처럼 염증세포의 치근단으로의 이행 및 그로 인한 골흡수를 막는 역할을 한다. 그러나 반복적인 지대주의 착탈은 임플란트 주위 연조직의 mucosal barrier를 파괴하여 골흡수를 야기할 수 있다. 만일 지대주의 반복적인 착탈 없이 수술 시에 보철 지대주를 채결한다면 골흡수의 양을 줄일 수 있을 것이다. 이는 기존의 수술방법으로는 한계가 있었으나 Cone Beam Computed Tomography(CBCT)를 이용한 guided surgery를 이용함으로써 임플란트 식립의 3차원적 정확성의 비약적인 향상으로 이런 술식이 가능하게 되었다. 본 증례는 정밀한 CBCT를 통한 분석 후 지르코니아 맞춤 지대주를 미리 제작하고 수술 시에 즉시 지대주와 임시보철물을 연결한 경우로 임상적으로 만족할만한 결과를 얻어 이를 보고하는 바이다.

• Tuberculosis and Respiratory Diseases
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• 제48권3호
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• pp.339-346
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• 2000

연구배경 : 혈액형 A 항원의 발현은 적혈구외에 비뇨기, 위장, 폐, 구강점막 동의 상피세포에도 존재한다. 조직 손상 치유 과정 중 인접 상피세포는 혈액형 A 항원이 소실되고, 상처가 치유되면 혈액형 A 항원은 다시 발현된다. 방광암등의 조직에서도 혈액형 A 항원이 소실될 경우가 발현유지 되는 경우보다 종양의 증식이 활발하다고 보고하였다. 또한 수술한 비소세포폐암의 조직에서 혈액형 A 항원이 소실될 경우 생존율은 불량하다고 보고하였다. 그러나, 이에 반하는 보고도 다수 있어 저자들은 근치적 절제술을 시행 받은 말초 혈액형이 A, AB형인 비소세포폐암에서 혈액형 A항원의 소실여부와 생존율과의 관계를 검색하였다. 대상 및 방법 : 원발성 비소세포폐암으로 진단 받은 후 근치 목적의 절제술을 받았던 환자 76명중 혈액형이 A형, 또는 AB형인 26명을 대상으로 paraffin에 보관된 조직을 면역조직화학염색법을 이용하여 혈액형 A항원의 소실유무을 확인한 후, 발현 및 소실에 따른 생존율은 Kaplan-Meier법, Log-rank로서 검색하였다. 결 과 : 대상군은 26례(A형 : 22례, AB형 : 4례)로 남 : 녀는 20 : 6이었고, 평균연령은 63세였으며, 조직학적으로 편평상피암 16례, 선암 6례, 대세포암 4례였고, TNM병기는 I 16례, II 5례, IIIA 5례였다. 종양조직에서 A항원 발현유지는 15례(58%), A 항원 소실은 11례(42%)였으며, A 항원 발현율과 병리조직형, 조직의 분화도와는 상관관계가 없었다. A항원 발현유지군과 소실군간의 중간 생존기간은 11개월, 18개월이며, 2년 생존률은 36%, 64%로서 A 항원 소실군의 예후가 양호하였으나, 통계적으로 유의한 차이는 없었다. 결 론 : 원발성 비소세포폐암으로 절제수술을 받았던 혈액형이 A형, 또는 AB형인 환자중 조직에서의 A 항원소실은 42%이었고, A항원 소실군이 발현유지군보다 생존율이 길었으나 예후인자로서의 유의성은 찾지 못했다.

• 대한기관식도과학회지
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• 제8권1호
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• pp.29-34
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• 2002

Background and Objectives : Sulfur dioxide gas is one of the major airborne Pollutants noxious to human in industrialized countries. The most vulnerable areas in the human respiratory system were the trachea and main bronchi and a gradient of decreasing damage was observed in the peripheral tracheobronchial tree. Induced functional alteration was increased mucosal permeability, and morphological changes were epithelial sloughing, intracellular edema, mitochondrial swelling, widened intercellular spaces, and ciliary cytoplamic extrusions. The laminins are a family of extracellular matrix glycoproteins localized in the basement membrane. Their primary role is cell-matrix attachment, but many additional biologic activities, including Promoting cell growth and migration, tumor growth and metastasis, wound repair, and graft survival, have been demonstrated. Materials and Methods : Histologic changes and expression of laminin in tracheal mucosa sacrificed at 1 day, 2 day, 3 day, 1 week, 2 weeks, and 3 weeks after continued SO2 exposure of 250 ppm for 30 minutes a day(to 7week) were studied in rats. In this study, mild immune reaction for laminin was noted at the apical cytoplasm of epithelial cells and basement membrane one day after a 7 week $SO_2$ exposure. The cilia and nucleoi of epithelial cells were normal and no immune reaction was noted in Goblet cells. The lamina propria of the tracheal tissue was infiltrated by monocytes and lymphocytes. Results : At 24 hours after exposure, all tracheal cells except Goblet cells revealed a mild immune reaction for laminin. No immune reactions were noted in the basement membrane. At 72 hours after exposure, mild or moderate immune reactions for laminin was seen in the tracheal cell cytoplasm. Irregular faint immune reaction for laminin was noted in the basement membrane. At 1 week after exposure, strong immune reaction for laminin was detected over all tracheal cells, and the basement membrane was seen clearly. At 2~3 weeks after exposure, strong immune reaction for laminin was seen in all tracheal epithelial cells except Goblet cells and a mild immune reaction was partly revealed in the basement membrane. Conclusion : Our study suggests that 502 produces histologic damage on the tracheal mucosa. Longer duration after exposure of $SO_2$ makes more progressive healing on the tracheal mucosa and increased immunoreactivity for laminin.

• Journal of Ginseng Research
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• 제38권3호
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• pp.167-172
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• 2014

Background: Roles of immune reaction and toll-like receptor-4 (TLR-4) have widely been established in the pathogenesis of alcoholic liver disease (ALD). Methods: We evaluated the biologic efficacy of Korean Red Ginseng (KRG), urushiol, and probiotics (Lactobacillus rhamnosus R0011 and Lactobacillus acidophilus R0052) in mouse models of ALD. Sixty C57BL/6 mice were equally divided into six feeding groups for 10 weeks: normal diet, alcohol, control, alcohol + KRG, alcohol + urushiol, and alcohol + probiotics. Alcohol was administered via a LiebereDeCarli liquid diet containing 10% alcohol. TLR-4 expression, proinflammatory cytokines, and histology, as well as the results of liver function tests were evaluated and compared. Results: No between-group differences were observed with regard to liver function. TLR-4 levels were significantly lower in the KRG, urushiol, and probiotics groups than in the alcohol group ($0.37{\pm}0.06ng/mL$, $0.39{\pm}0.12ng/mL$, and $0.33{\pm}0.07ng/mL$, respectively, vs. $0.88{\pm}0.31ng/mL$; p < 0.05). Interleukin-$1{\beta}$ levels in liver tissues were decreased among the probiotics and KRG groups. The tumor necrosis factor-${\alpha}$ level of liver tissue was decreased in the KRG group. Conclusion: The pathological findings showed that alcohol-induced steatosis was significantly reduced by KRG and urushiol. As these agents improve immunologic capacity, they may be considered in potential anti-ALD treatments.

• 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).

• 대한치과교정학회지
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• 제34권2호
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• pp.143-152
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• 2004

하악과두 연골이 발생하고 분화하는 과정에서 나타내는 특성을 규명하기 위하여, 발생 16, 18일과 출생 1일, 5일, 10일, 20일 및 30일 후의 ICR생쥐의 하악과두를 형태학적으로 분석하고, 생쥐 I형, II형, X형 교원질, Indian hedgehog (IHH) 및 BMP-4 등의 mRNA 발현을 in-situ hybridization 방법으로 연구하였다. 1. 생쥐 I형 및 II형 교원질 mRNA는 하악과두의 발생 및 성장과정에서 모두 발현되었다. I형 교원질 mRNA는 휴지층과 증식층의 상부에서 관찰된 반면 II형 교원질은 휴지층과 증식층 그리고 비대연골층의 상부에서 관찰되었다. 2. 하악과두 연골은 성장에 따라 비대연골층이 계속 증가하는 소견을 보였으며, 비대 연골층의 세포들은 특징적으로 X형 교원질 mRNA의 발현을 보였다. 3. BMP-4 mRNA는 하악과두 연골 원기와 골화중인 하악골체에서 모두 발현되었다. 4. IHH mRNA는 하악과두의 발생과정에서 증식 연골층의 하부와 비대연골층의 상부에서 선택적으로 관찰되었다.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.