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토끼 모델에서 탈미네랄화된 골분을 함유한 PLGA 지지체를 이용한 조직공학적 골 재생
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.
DBP스폰지에서 BMP-2의 효과확인 및 섬유륜 조직재생
최진희 ( Jin Hee Choi ),장지욱 ( Ji Wook Jang ),김순희 ( Soon Hee Kim ),홍희경 ( Hee Kyung Hong ),민병현 ( Byung Hyun Min ),손영숙 ( Youngsook Son ),이종문 ( John M Rhee ),강길선 ( Gilson Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4
Demineralized bone particle(DBP) that affects to cell proliferation and differentiation has been used as biomaterials. In this study, we evaluated 3-dimensional DBP sponge and Collagen sponge on proliferation and phenotype maintenance of annulus fibrosus cells. DBP sponge were prepared by freeze-drying method after addition 2 wt% DBP solution. sponge was crosslinked with 1-ethyl-(3-3-dimethyl aminopropyl) carbodiimide hydrochloride(EDC) solution with 50 mM concentration for 24 hrs and lyophilized. We seeded cells in DBP sponge and Collagen sponge. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. DBP sponge and Collagen sponge were characterized by scanning electron microscopy(SEM). Reverse transcription polymerase chain reaction(RT-PCR) was assessed to measure mRNA expression for type I collagen, type II collagen, aggrecan and osteonectin and glycosaminoglycan(GAG) quantity from annulus fibrosus cells in sponges. In MTT assay result, DBP sponges were higher cell viability. In SEM observation, we observed that DBP sponge has uniform porosity. In addition, annulus fibrosus stronly expressed their specific mRNA. and produced well sGAG in DBP sponge. This result indicates that DBP sponge is useful for intervertebral disc regeneration.
유화동결건조법으로 제조된 HA/PLGA 지지체를 이용한 연골 재생
정수현 ( Su Hyun Jung ),장지욱 ( Ji Wook Jang ),김순희 ( Soon Hee Kim ),홍현혜 ( Hyun Hye Hong ),오아영 ( A Young Oh ),이종문 ( John M. Rhee ),강영선 ( Young Sun Kang ),강길선 ( Gil Son Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4
Adult articular cartilage tissue has poor capability of self-repair. Therefore, a variety of tissue engineering approaches are motivated by the clinical need for articular repair. PLGA and hyaluronic acid(HA) has been widely used as biocompatible scaffolds materials to regenerate tissue. HA loaded PLGA scaffolds were prepared by a emulsion freeze-drying method. The chondrocytes were seeded on the HA-PLGA scaffolds and measured by MTT assay. Morphological observation, histology, biological assay for collagen and sGAG, and PCR were performed. In MTT assay result, scaffolds containing HA were higher cell viability then only PLGA scaffolds. Although collagen, sGAG, mRNA and collagen type II were greater than HA-PLGA scaffolds and collagen type I was less than HA/ PLGA scaffolds. When we cultured cartilage cell of rabbit in vitro, we observed better to keep the characteristic of cartilage cell in the HA-PLGA scaffolds than that PLGA scaffolds. This study suggests that HA/PLGA scaffold may serve as a potential cell delivery vehicle and a structural basis for in vitro tissue engineered articular cartilage.