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IGF-I이 배양된 연골세포의 콜라젠 합성에 미치는 영향
한상훈,조종한,이종환,홍혜남,김승후,이재담 大韓成形外科學會誌 2000 Archives of Plastic Surgery Vol.27 No.1
Cartilage is one of the most commonly manipulated tissue in esthetic and reconstructive surgery. Cartilage has an important role in longitudinal bone growth. Anabolic hormones and locally produced peptide growth factors are known to influence this process Matrix composition changes through proliferation, maturation, and differentiation of chondrocytes, and endochondral ossification thereafter. Defined cartilage matrix is synthesized during the maturation of chondrocytes where the major change is the increment of type Ⅱ collagen. Variable sulfated mucololysaccharides and hyaluronic acid are also synthesized during this maturation. IGF-I(insulin like growth factor-I), so called somatomedin C, is a prominent growth factor in serum. IGF-I is known to be involved in long growth. IGF-I is affected by pituitary growth hormone. There are few studies done on IGF-I effect in cartilage matrix formation and possible changes of collagen subtypes. This experiment was designed to see the IGF-I effect on the colagen synthesis of cultured chondrocytes. Optimal concentration of IGF-I for the experiment was determined using H3-thymidine incorporation into DNA. The IGF-I effect on collagen synthesis was studied using H3-proline. The IGF-I effect on the synthesis of subtypes of collagen was studied using SDS-PAGE and immunocytochemical staining. Chondrocytes were isolated from the ears of New Zealand white rabbit and cultured in 2 X 10??cells/300㎍ density. IGF-I increased DNA synthesis, and optimal concentration of IGF-I was determined by dose-relationship curve as 10ng/ml. Collagen synthesis was increased by IGF-I. Type Ⅱ collagen was increased on SDS-PAGE with IGF-I and this gel electrophoresis showed thpe X collagen, also. The increase in type Ⅱ collagen was confirmed with immunocytochemical staining, the reaction becoming stronger with the addition of IGF-I. Type Ⅰ collagen was not changed with IGF-I on immunocytochemistry. We conclude that IGE-I is an important modulator influencing not only proliferation and maturation but also terminal different-iation of chondrocytes.
조종한,윤근철,문 경,이재담 大韓成形外科學會 1996 Archives of Plastic Surgery Vol.23 No.3
A cartilage is the most commonly used tissue in reconstructive and esthetic surgery as an implant and it is closely related with ossification process. There were few reports of primary culture of chondrocytes in Korea because of complicated and time consuming procedures, and frequent fibroblast contamination. These previous reports were limited to monolayer cell culture, so no report exists about high density three dimensional primary culture of rat chondrocytes. We performed primary culture of rat chondrocytes using growth cartilage at the costochondral junction in young male Sprague-Dalwey rat of 80-120 gram body weight and culturing in 6.7×10⁴/360μl cell density with stainless steel Peni-cylinder in the center of the 35mm culture dish. Characterization with inverted microscopy, toluidine blue staining, and measuring alkaline -phosphatase activity was performed. To evaluate the effects of bovine pituitary extract (BPE) and fibroblast growth factor (FGF) on chondrocyte growth, the BPE or the FGF was added to the culture media. BPE is known to promote cell growth, and the FGF is the known active component of BPE. [³H]- thymidine incorporation was used as an index of DNA synthesis during cell proliferation. With adding the BPE or the FGF to the culture media, increased DNA synthesis was defected in comparison with the control. The effect of the BPE may be partially contributed by the FGF because the FGF is one of the active components of the BPE. To evaluate effect of the FGF,[³H]-proline incorporation was used as an index of collagen synthesis. At the level of 0.01ng/ml of the FGF concentration, the amount of collagen synthesis was almost same with the control, but it decreased with increasing the FGF concentration above 0.1ng/ml. From these data, we found that the FGF stimulates growth of chondrocytes and inhibits differentiation of chondrocytes. We consider that it can modulate ossification process with these actions.
토기의 귀 연골 세포 배양에서 여러 가지 성장 인자의 효과
한상훈,강정훈,성창기,이재담 大韓成形外科學會 1997 Archives of Plastic Surgery Vol.24 No.5
Cartilage is used frequently as a graft material in plastic & reconstructive surgery. Nowadays, many investigations such as allograft of cartilage, effects of various growth factors in cultured chondrocytes, and cartilaginous tissue replacement for bony defects are being performed in many countries. In this study, we isolated chondrocytes from ear cartilage of the New Zealand white rabbits weighing 200 to 250 g. We performed high density culture using penicylinder and observed the effect of various growth factors on cell proliferation(DNA synthesis), protein synthesis(leucine uptake), collagen synthesis(proline uptake) and matrix secretion(glycosaminoglycan synthesis). Optimal concentrations of the growth factors, were determined from dose-response studies. The transforming growth factor(TGF)-β and basic fibroblast growth factor(bFGF) administered on the 3 day of culture showed the effect to increase the DNA synthesis. TGF-βand bFGF showed additive action when administered together in high density culture and showed synergistic action in lower density culture. And bFGF showed inhibitory effect in matrix synthesis when administered alone or with TGF-β, which means bFGF has dedifferentiation effect. The chondrocytes culture technique using the rabbit ear cartilage was established in this study and administration of both TGF-βand bFGF was found to be very helpful in the chondrocyte culture.
정호균,한상훈,성창기,윤근철,이재담 大韓成形外科學會 1996 Archives of Plastic Surgery Vol.23 No.6
Cartilage is very useful as structural support tissue for cosmetic repair in plastic and reconstructive surgery. In vitro culture of chondrocytes is a basis for histological and biochemical studies of cartilage. Since the chondrocytes culture was reported, various culture methods have been developed. Thereafter gradually high-density, three-dimensional culture methods suitable for clinical purposes have been performed. Various methods of isolation and culture of the chondrocytes were reported and the biochemical characteristics of the cultured chondrocytes have been reported different according to the experimental animals or cartilage types. In this study we present isolation and culure method of the chondrocytes using the rabbit ear and rib cartilage, and the cells were characterized microscopically and biochemically. The results of this study are as follows. 1. The adequate weigh of New Zealand White rabbit for chondrocyte culture was 200 to 250 gm. 2. The cells were polygonal in morphology and were viable under the inverted microscope. 3. The cells were characterized as chondrocytes because they show alkaline phosphatase activity and metachromasia in toluidine blue staining. 4. The cells proliferation was identified in high density culture using penicylinder.
임상연구 : Remifentanil과 Lidocaine이 Sevoflurane 마취유도 시 기관내삽관에 필요한 흡입시간에 미치는 영향
이재담 ( Jae Dam Lee ),정창영 ( Chang Young Jeong ),최정일 ( Jeong Il Choi ),이형곤 ( Hyung Gon Lee ),정성태 ( Sung Tae Chung ),김웅모 ( Woong Mo Kim ) 대한마취과학회 2008 Korean Journal of Anesthesiology Vol.55 No.5
Background: This study was conducted to investigate the optimal time interval for tracheal intubation and the effect of adjuvant drugs such as remifentanil and lidocaine during induction and tracheal intubation using sevoflurane inhalation without muscle relaxant. Methods: This study enrolled patients with the age of 20-60 years old and ASA 1 or 2. Patients were randomly assigned into one of 4 groups (S, SR, SRL, SL), in which they were given remifentanil (R) i.v. at a rate of 0.25 μg/kg/min, or lidocaine (L) i.v. bolus of 1.5 mg/kg during sevoflurane inhalation (S). Anesthesia was performed as inhalation induction 2 minutes after pre-filling with sevoflurane 8 vol%. The time interval between induction and tracheal intubation was determined using up-and-down method. When satisfied all of the categories of response to tracheal intubation, the case was assigned to `success`, otherwise `fail`. In each groups, effective time for successful intubation in 50% (ET50) and 95% (ET95) were calculated by probit analysis. Results: ET50 was 3.90 minutes (95% confidence interval 3.32-4.38) in group S, 3.18 minutes (2.92-3.48) in group SL, 2.83 minutes (2.47-3.07) in group SR, and 2.68 minutes (2.37-2.95) in group SRL. In group S, SL, SR, and SRL, ET95 was 4.52 minutes (4.17-7.95), 3.63 minutes (3.37-4.97), 3.30 minutes (3.06-4.64), and 3.12 minutes (2.89-4.42), respectively. Conclusions: The optimal time to intubate successfully using sevoflurane without muscle relaxant in 95% patients was 4.5 minutes. The optimal time is reduced to 3.1 minutes by coadministration of remifentanil and lidocaine. (Korean J Anesthesiol 2008;55:565~9)