간헐적 정수압이 세포의 부착에 미치는 영향성 평가

김영직(Young Jick Kim),이명곤(Myung-Kon Lee),박수아(Su A Park),신호준(Ho Joon Shin),김인애(In Ae Kim),이용재(Yong Jae Lee),신지원(Ji Won Shin),신정욱(Jung-Woog Shin) 한국정밀공학회 2005 한국정밀공학회 학술발표대회 논문집 Vol.2005 No.10월

In this study, effects of IHPs with various resting times to cell adhesion were investigated through the measurements of cell adhesive force, number and area of focal contacts (stained vinculin spots), and projected cell area, perimeter and circularity. In addition correlation tests and curve estimations using the experimental results were performed for the finding an essential factor for increment of cell adhesive force. In the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). Average projected area and perimeter of cells at Group 5 were increased significantly (p<0.05), while average circularity of cells at Group 5 incubated for 210 minutes was decreased significantly (p<0.05). In the digital image analysis of focal contacts containing vinculins, area and numbers of focal contacts per cell at Group 5 were higher than those of the other groups. This study indicated that IHP with appropriate resting time could contribute in improving cell adhesive force, cell spreading, development of cytoskeleton and formation of focal contacts. And cell adhesive force was correlated to the morphological aspects of cell and development of focal contacts. Particularly, area of focal contacts was closely related to cell adhesive force.

간헐적 정수압의 다양한 패턴에 따른 세포 부착력과 F-actin 및 vinculin 의 분포 변화 연구

김영직(Young Jick Kim),박수아(Su-A Park),신호준(Ho Joon Shin),김인애(In Ae Kim),이용재(Yong Jae Lee),허수진(Su-Jin Heo),황영미(Young-Mi Hwang),신정욱(Jung-Woog Shin) 한국정밀공학회 2004 한국정밀공학회 학술발표대회 논문집 Vol.2004 No.10월

Cell adhesion to any material surface is considered to be fundamental and important phenomenon in the fields of tissue engineering. Cell adhesion molecules, mechanism, and attachment force have been studied and described a lot. However, the effects of mechanical stimuli on the adhesive forces still have been left much to be investigated. In this study, to investigate the changes in cell adhesive force due to resting time period during the intermittent hydrostatic pressurizing (IHP), cells were cultured under the IHP with various resting times. Then the cell adhesive forces were measured quantitatively utilizing a cell detachment test system and immunofluorescent staining was performed using fluorescent microscopy. In the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). The results indicated that IHP can contribute in improving cell adhesive force and some of time intervals were required for the expression of cell response.

세포 내 칼슘 농도의 변화에 따른 간헐적 정수압이 세포 부착력에 미치는 영향

김동화(Dong Hwa Kim),김영직(Young Jick Kim),신지원(Ji Won Shin),신정욱(Jung-Woog Shin) 대한기계학회 2008 대한기계학회 춘추학술대회 Vol.2008 No.11

We investigated the effects of intermittent hydrostatic pressure with various duration of resting period on changes in calcium (Ca²?) concentration and adhesive forces of cells on substrates. The quantitive adhesive forces of cells were measured under various resting periods. When the pressure applied to the cells, the concentration of Ca²? increased. Under intermittent hydrostatic pressure, the concentration of Ca²? was maintained under a resting period of 15 min, while it was not decreased with other resting periods of less than 15 min, With a resting period of 15 min, the magnitudes of adhesive forces were significantly increase. In addition, the adhesive forces were measured with and without Ca²? chelating agents to evaluate the effect of Ca²? on cell adhesiveness. When Ca²? ions were chelated, the adhesive forces dramatically decreased, even under intermittent hydrostatic pressure. We conclude that Ca²? plays an crucial role in modulating the adhesive forces of cells, and that the concentration of Ca²? can be increased by intermittent hydrostatic stimuli

기계적 자극이 다층 구조의 나노파이버 지지체의 평활근 세포에 미치는 영향

신지원,김동화,허수진,김수향,김영직,신정욱,Shin, Ji-Won,Kim, Dong-Hwa,Heo, Su-Jin,Kim, Su-Hyang,Kim, Young-Jick,Shin, Jung-Woog 대한의용생체공학회 2008 의공학회지 Vol.29 No.1

The object of this study is to investigate the effects of intermittent cyclic stretching on the smooth muscle cells (SMCs) seeded onto aligned multi-layered fibrous scaffold. To make multi-layered fibrous scaffold, polyurethane (PU) and poly(ethylene oxide) (PEO) were electrospun alternatively, then were immersed into distilled water to extract PEO. Various types of scaffolds were fabricated depending on fiber directions, i.e., aligned or randomly oriented. The direction of stretching was either parallel or vertical to the fiber direction for the aligned scaffolds. The stretching was also applied to the randomly aligned scaffolds. The duration of stretching was 2 min with 15 min resting period. During the stretching, the maximum and minimum strain was adjusted to be 10 and 7%, respectively with the frequency of 1 Hz. The bioactivities of cells on the scaffolds were assessed by quantifying DNA, collagen, and glycosaminoglycan (GAG) levels. And the cell morphology was observed by staining F-actin. SMCs under parallel stretching to the fiber direction responded more positively than those in other conditions. From the results, we could explain the morphological effect of a substrate on cellular activities. In addition the synergistic effects of substrate and mechanical stimuli effects were confirmed.

기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증

이용재,신지원,신호준,김찬환,박기동,배진우,서형연,김영직,신정욱,Lee, Yong-Jae,Shin, Ji-Won,Shin, Ho-Jun,Kim, Chan-Hwan,Park, Ki-Dong,Bae, Jin-Woo,Seo, Hyoung-Yeon,Kim, Young-Jick,Shin, Jung-Woog 대한의용생체공학회 2007 의공학회지 Vol.28 No.4

The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

감마선을 이용한 돼지 무세포 연골세포외기질이 함유된 CMC 수화젤의 제조 및 특성 연구

정성린(Sung In Jeong),박종석(Jong-Seok Park),권희정(Hui-Jeong Gwon),안성준(Sung-Jun An),임윤묵(Youn-Mook Lim),송보람(Bo Ram Song),김영직(Young Jick Kim),민병현(Byoung-Hyun Min),김문석(Moon Suk Kim) 한국고분자학회 2017 폴리머 Vol.41 No.1

본 연구는 감마선을 이용하여 가교된 유착방지용 carboxymethyl cellulose sodium salt(CMC-Na)와 porcine cartilage acellular matrix(PCAM) 수화젤을 제조하였다. CMC는 높은 농도의 수용액에서 감마선에 의해 가교가 되는 천연고분자 중에 하나이다. 방사선 가교된 CMC/PCAM 수화젤의 물리/화학적 특성을 알기 위해 SEM, 젤화율, 젤강도, 팽윤도, FTIR-ATR 등 분석을 통해 특성을 확인하였다. 감마선으로 가교된 CMC/PCAM 수화젤에서 PCAM 비율이 증가할수록 젤화율과 젤강도는 감소하였고 팽윤도는 증가하였다. 또한 CMC/PCAM 수화젤에서 세포의 부착도와 증식도가 CMC 수화젤에 비해 낮음을 확인하였다. 결론적으로, 본 연구에서는 방사선으로 가교된 CMC/PCAM 수화젤은 유착방지를 위한 유착방지제로서 응용이 기대된다. In this study, a novel crosslinked carboxymethyl cellulose sodium salt (CMC) and porcine cartilage acellular matrix (PCAM) hydrogel for adhesion prevention were prepared by gamma-irradiation. CMC is known as a kind of cross-linked natural polymer at highly concentrated aqueous solution under action of gamma-irradiation. The radiationcrosslinked CMC/PCAM hydrogels were characterized by scanning electron microscope (SEM), gel-fraction, gel strength, swelling behavior, and FTIR-ATR spectroscopy. The results showed that the PCAM addition increased the swelling behavior and decreased the gel-fraction and gel strength of CMC/PCAM hydrogels. The degree of attachment and proliferation of human fibroblast on CMC/PCAM hydrogel was lower than the CMC hydrogels. In conclusion, this study suggests that the radiation cross-linked CMC/PCAM hydrogels may be useful as an anti-adhesion barrier.

방사선 가교된 유착방지용 Carboxymethyl Cellulose/Porcine Cartilage Acellular Matrix 수화젤 필름의 물리적 특성 및 부착 방지 평가

정성린 ( Sung In Jeong ),박종석 ( Jong-seok Park ),권희정 ( Hui-jeong Gwon ),안성준 ( Sung-jun An ),송보람 ( Bo Ram Song ),김영직 ( Young Jick Kim ),민병현 ( Byoung Hyun Min ),김문석 ( Moon Suk Kim ),임윤묵 ( Youn-mook Lim ) 한국화학공학회 2017 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.55 No.1

본 실험은 방사선을 이용하여 유착방지용 Carboxymethyl cellulose sodium salt (CMC)/Porcine Cartilage Acellular Matrix (PCAM) 수화젤 필름에 대해 연구하였다. 방사선 조사시 CMC/PCAM의 농도 및 혼합비율에 따른 필름의 형태학적 구조, 젤화율, 젤강도, 팽윤도 등을 분석하였다. 방사선에 의해 가교된 CMC/PCAM 필름은 CMC 필름보다 물리적인 특성인 젤화율이 낮게 측정되었다. 또한, 가교된 CMC 필름보다는 CMC/PCAM 필름에서 인간 혈관내피세포의 부착 및 증식율이 감소하였다. 우리는 PCAM에 함유한 세포의 항부착 성분이 도입된 CMC/PCAM 필름과 CMC 필름은 세포의 접착 및 증식율을 낮추는 것을 확인하였다. 결론적으로, 본 연구에서는 방사선 가교된 CMC/PCAM 수화젤 필름은 유착방지 향상을 위한 유착방지제로서 응용이 기대된다. In this study, intermolecular crosslinked carboxymethyl cellulose sodium salt (CMC) and porcine Cartilage Acellular Matrix (PCAM) blended hydrogel films for anti-adhesive barriers were prepared by gamma-ray radiation. The effects of the CMC/PCAM concentration and blending ratio on the morphology, gel fraction, gel strength, and degree of swelling were determined. The results indicated that crosslinked CMC/PCAM films show significantly lower the gel-fraction than CMC films. The degree of attachment and proliferation of human vascular endothelial cells on CMC/PCAM films was lower than the CMC films. We show the capacity of the CMC and PCAM to be hydrogel films, and the ability to reduce cell adhesion and proliferation on these films by modification with cell anti-adhesion molecules of PCAM. In conclusion, this study suggests that radiation cross-linked CMC/PCAM hydrogel films endowed with anti-adhesion ligands may allow for improved regulation of cell anti-adhesion behavior for prevent peritoneal adhesions.

신경세포의 Outgrowth 향상을 위한 마이크로 파이버 지지체와 전단응력의 영향

김인애,박수아,김영직,김수향,신호준,이용재,신지원,신정욱 한국공작기계학회 2005 한국공작기계학회 춘계학술대회논문집 Vol.2005 No.-

We introduced mechanical stimuli and micro patterned substrate with micro fibers to investigate the effects of those on neurite outgrowth along with nerve growth factor (NGF) in vitro. Micro fiber substrates were fabricated using an electros pinning process. And PC-12 cells cultured on substrates were simulated with newer growth factor and laminar flow shear stress in a fluid flow system. The results suggest that micro fiber substrates and fluid-induced shear stress are promising for simulating neuronal regeneration in a desired direction.

정중운동신경과 척골운동신경의 전기생리학적 연구

김종순,이현옥,안소윤,구봉오,남건우,김영직,김호봉,류재관,류재문 대한정형도수치료학회 2005 대한정형도수물리치료학회지 Vol.11 No.2

The determination of peripheral nerve conduction velocity is and important part to electrodiagnosis. Its value as neurophysiologic investigative procedure has been known for many years but normal value of median and ulnar motor nerve was poorly reported in Korea. To evaluate of median and ulnar motor nerve terminal latency, amplitude of CMAP(compound muscle action potential), conduction velocity and F-wave latency for obtain clinically useful reference value. 71 normal volunteers(age,19-65 years;142 hands) examined who has no history of peripheral neuropathy, diabetic mellitus, chronic renal failure, endocrine disorders, anti-cancer medicine, anti-tubercle medicine, alcoholism, trauma, radiculopathy. Nicolet Viking Ⅱ was use for detected terminal latency, amplitude of CMAP, conduction velocity and F-wave latency of median and ulnar motor nerve. Data analysis was performed using SPSS. Descriptive analysis was used for obtain mean and standard deviation, independent t-test was used to compare between Rt and Lt side also compare between different in genders. The results are summarized as follows: 1. Median motor nerve terminal latency was right 3.00ms, left 2.99ms and there was no significantly difference between right and left side and genders. 2. Median motor nerve amplitude of CMAP was right 17.26mV, left 17.50mV and there was no significantly differences between right and left side and genders. 3. Median motor nerve conduction velocity was right 57.89m/sec, left 58.03m/sec and there was no significantly difference between right and left side and genders. 4. Median motor nerve F-wave latency was right 25.74ms, left 25.59ms and there was significantly differences between genders. 5. Ulnar motor nerve terminal latency was right 2.38ms, left 2.45ms and there was significantly differences between right and left side. 6. Ulnar motor nerve amplitude of CMAP was right 15.99mV, left 16.02mV and there was no significantly differences between right and left side and genders. 7. Ulnar motor nerve conduction velocity was right 60.35m/sec, left 59.73/sec and there was no significantly differences between right and left side and genders. 8. Ulnar motor nerve F-wave latency was right 25.53ms, left 25.57ms and there was significantly differences between genders.