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김선경 ( Sun Kyung Kim ),홍금덕 ( Keum Deok Hong ),장지욱 ( Ji Wook Jang ),이상진 ( Sang Jin Lee ),김문석 ( Moon Suk Kim ),강길선 ( Gil Son Khang ),이일우 ( Il Woo Lee ),이해방 ( Hai Bang Lee ) 한국조직공학과 재생의학회 2004 조직공학과 재생의학 Vol.1 No.2
Bone marrow stromal stem cells (BMSCs) were harvested from the femurs and tibias of adult female Fischer rat. BMSCs were suspended at 2 × 107 cell/mL in a 25% Pluronic F127 (PF127) solution and then polyglycolide (PGA) nonwoven was saturated with 50 μl PF127 solution. The ability of the PGA/PF127 with BMSCs to be integrated and to promote nerve regeneration in the transected rat spinal cord was investigated. Fischer rat received an implant consisting of the BMSCs suspended in the PGA/PF127 in the gap (T8~T9;~5 mm) created by the spinal cord resection. For histological evaluation, the implants were removed after 4 and 8 weeks. Thin sections were cut from paraffin embedded tissue and histological sections were stained hematoxylin and eosin staining. Motor functional outcome measurements using the Basso-Beattie-Bresnehan (BBB) score performed weekly to 8 weeks postinjury. It was observed that the effects of the PGA/PF127 with BMSCs on neuroinduction (Group III, tissue engineered) are stronger that PGA/PF127 without BMSCs (Group II, matrix control) and blank control (Group I). In conclusion, these results suggest that BMSCs may have an important role for spinal cord regeneration of tissue engineering area.
이시우 ( Shi Woo Lee ),허수진 ( Su Jin Heo ),장지연 ( Ji Yeon Jang ),정재영 ( Jae Young Jeong ),김수향 ( Su Hyang Kim ),박수아 ( Su A Park ),전은수 ( Eun Su Jeon ),신정욱 ( Jung Woog Shin ) 한국조직공학과 재생의학회 2010 조직공학과 재생의학 Vol.7 No.1
The objective of this study was to investigate the efficacy of a newly designed perfusion bioreactor which provides flow-induced mechanical stimulation on the cells residing in the intra-morphology controllable scaffolds. For this, we fabricated scaffolds composed of poly e-caprolactone (PCL) and micro-sized hydroxyapatite (HA) particles using rapid-prototyping process. Also a new bioreactor system for perfusion culture was designed and developed. For the analyses of cellular responses of bone-tissue related cells cultured in the perfusion bioreactor system, various biological assays were performed such as MTT test, DNA content measurement, FE-SEM and live/dead staining. The cells used in this study were MG-63 (human osteoblast-like cell line) and mesenchymal stem cells from New Zealand white rabbits. Our results showed that the cells cultured by the perfusion bioreactor resulted in higher proliferation rate and mineralization of extracellular matrices than those cultured in static culture. From this study, we could confirm the potentials of 1) a newly developed perfusion bioreactor, 2) intra-morphology controllable scaffolds composed of PCL and HA particle and 3) the combination of the suggested scaffolds and perfusion culturing system in relation to bone tissue engineering.
정수현 ( Su Hyun Jung ),김순희 ( Soon Hee Kim ),양재찬 ( Jae Chan Yang ),홍현혜 ( Hyun Hye Hong ),김혜린 ( Hye Lin Kim ),김원 ( Won Kim ),손영숙 ( Young Sook Son ),( Sang Jin Lee ),( Mark Van Dyke ),( James J Yoo ),이종문 ( 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.1
Articular cartilage that is difficult to recovery when damaged needs to tissue engineering. keratin are the intermediate filament proteins that form a dense meshwork of filaments throughout the of cells and generally expressed in particular pairs of type I and type II keratin proteins in a-specific and cellular differentiation-specific manner. In this study, we are developing an alternative approach that consists of generating chondrocytes anchored to poly(L-lactide-co-glycolide)(PLGA) scaffolds impregnated keratin(keratin/PLGA) using tissue-engineering principles. We prepared PLGA and keratin/PLGA scaffolds using solvent casting/salt leaching method. Chondrocytes were isolated from the articular cartilage of New Zealand white rabbit and cultured With DMEM/Ham`s F-12 supplemented with 10 % FBS, 1 % penicillin streptomycin, 200 mM L-glutamin, 50 ?g/ml of ascorbic acid and 15 mM HEPES buffer 1M. After 2weeks of cell seeding, we implanted keratin /PLGA scaffolds on the back of nude mice. Morphological observation, histology, biological assay for collagen and sGAG, and PCR were performed at each time point 1, 2, 3 and 6 weeks. The cell viability and the quantity of collagen and sGAG were better keratin/PLGA scaffolds than PLGA scaffolds. Specific mRNA, type II and type I collagen, for chondrocyte expressed significantly highly in keratin/PLGA scaffold. keratin/PLGA scaffold promotes in vivo chondrocyte of rabbit articular chondrocytes. This study suggests that keratin/PLGA scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered articular cartilage.
정수현 ( Su Hyun Jung ),김순희 ( Soon Hee Kim ),김초민 ( Cho Min Kim ),홍현혜 ( Hyun Hye Hong ),전나리 ( Na Ri Jeon ),김원 ( Won Kim ),이동원 ( Dong Won Lee ),이종문 ( John M Rhee ),강길선 ( Gil Son Khang ) 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.4
Nervous tissue engineering in combination with other therapeutic strategies is an emerging trend for the treatment of different CNS disorders and injuries. We proposed to use poly(L-lactide-co-glycolide)(PLGA) nerve channel impregnated demineralized bone particle(DBP) using tissue-engineering principles for the repair of spinal cord injury. We prepared DBP/PLGA nerve channel using ice particle-freeze drying method. Schwann cells(SCs), olfactory ensheathing cells(OECs) and bone marrow stromal cells(BMSCs) were seeded on DBP/PLGA nerve channel. The spinal cord was completely transected horizontally at two levels(T7 and T8), and DBP/PLGA nerve channel seeded cells was implanted in the lesion. For histological evaluation, the implants were removed after 2, 4 and 8 weeks and stained hematoxylin and eosin staining. Motor functional outcome measurements using the BBB scoring, sensory test and motor functional recovery test performed every week for 8 weeks post injury. It was observed that the effects of the DBP/PLGA nerve channel with cells(SCs, OECs and BMSCs), specially seeded SC on neuroinduction are stronger that DBP/PLGA nerve channel without cells and Blank control. In conclusion, these results suggest that SCs and DBP/PLGA nerve channel may have an important role for spinal cord regeneration of tissue engineering area.
Porcine small intestine submucosa(SIS), mainly composed of collagen and glycosaminoglycan, has been widely used as a material for organ and tissue reconstruction without immuno-rejection responses. Chemicallycrosslinked SIS sponges were prepared and characterized for bio-interactive wound dressings and tissue engineered scaffolds. SIS powder were reacted in aqueous solution of 3% acetic acid and 0.1% pepsin for 48 hrs and the prepared SIS solution was poured into mold and fabricated by freeze-drying method. The crosslinking reaction was performed using 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride(EDC) solution(distilled water(D.W.) : ethanol = 5 : 95) of 50, 100 and 200 mM concentration for 24 hrs. Then 1 wt% alginate solution was pass through the SIS sponge and lyophilzed. 102 mM calcium chloride(CaCl2) was used as crosslinking agent. The prepared sponges were characterized by scanning electron microscopy(SEM). The biocompatibility of NIH/3T3 fibroblast with these SIS sponges was found to be acceptable at a cell density of 2×104 cell/cm2. We investigated biodegradation of SIS sponge, MTT assay and H&E stain. From the MTT assay results, the biocompatibility of the crosslinked SIS sponge with NIH/3T3 fibroblast was confirmed. It was confirmed that SIS consists of organic components like tissue and showed inter-connective pores for adhesion and growth of cell. As a result of this study, we suggest that crosslinked alginate-SIS sponges fulfil many critical elements desirable in the application of wound dressing and tissue regeneration material.
장지욱 ( Ji Wook Jang ),박기숙 ( Ki Suk Park ),김순희 ( Soon Hee Kim ),박종수 ( Chong Soo Park ),김문석 ( Moon Suk Kim ),한창환 ( Chang Whan Han ),이종문 ( John M. Rhee ),강길선 ( Gil Son Khang ),이해방 ( Hai Bang Lee ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.1
Demineralized bone particle (DBP), one of the significant natural bioactive materials, has a powerful inducer of new bone growth. It has been recognized that DBP contains many kinds of osteogenic and chondrogenic cytokines as bone morphogenic protein (BMP). BMP acts as local mitogen to stimulate proliferation of mesenchymal stem cell. Bone marrow stromal cell (BMSCs) can be differentiated in culture into osteoblasts, chondrocytes, and myoblasts with controlling of the environment of cell growth. In this study, we developed the DBP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. PLGA/DBP scaffolds were prepared by solvent casting/salt leaching method and characterized by porosimeter, scanning electron microscopy, and differential scanning calorimetry. BMSCs were isolated from femur of New Zealand white rabbit and cultured with osteo-medium for osteogenic differentiation. DBP impregnated PLGA scaffolds with BMSCs were implanted into the head and femur of New Zealand white rabbit to observe the effect of DBP on the osteoinduction compared with control scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were stained hematoxylin & eosin and safranin-O. It can be observed that the porosity was above 94.6% and the pore size was above 69.8 ?m. In vivo study, we could observe that similar to bone tissue region in PLGA/BMSCs and PLGA/BMSCs/DBP groups, but bone tissue region did not occur almost in control scaffolds. From these results, it seems that DBP play an important role for bone induction on BMSCs.