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인간 디스크세포와 PLGA/DBP 지지체를 이용한 생체조직공학적 바이오 디스크
고연경 ( Youn Kyung Ko ),김순희 ( Soon Hee Kim ),하현정 ( Hyun Jung Ha ),김문석 ( Moon Suk Kim ),한창환 ( Chang Whan Han ),손영숙 ( John M. Rhee ),이종문 ( Youngsuk Son ),이해방 ( Hai Bang Lee ),강길선 ( Gilson Khang ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.1
We fabricated poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle( DBP)(PLGA/DBP) to investigate the effect of cell viability, proliferation and characteristic maintenance of intervertebral disc(IVD) cells. DBP-loaded PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in PLGA/DBP scaffold, and then cell viability and proliferation according to DBP content were analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide(MTT) assay. In MTT assay results, cell viability in scaffolds impregnated 20 and 80 wt% of DBP were higher than other scaffolds. Reverse transcript polymerase chain reaction(RT-PCR) was assessed to measure mRNA expression of the disc target genes(aggrecan and type II collagen) of human IVD cells about various content of DBP. After implantation, thin sections were cut from paraffin embedded tissues and histological sections were stained H&E staining for observation of disc cell distribution. We concluded that the using of DBP(especially 20 wt% of DBP) in terms of scaffold fabrication for bio-disc with human IVD cells helps growth of disc cells maintained their phenotypes.
하현정 ( Hyun Jung Ha ),김순희 ( Soon Hee Kim ),윤선중 ( Sun Jung Yoon ),고연경 ( Youn Kyung Ko ),이은경 ( Eun Kyung Lee ),손영숙 ( Young Sook Son ),김문석 ( Moon Suk Kim ),이종문 ( John M. Rhee ),강길선 ( Gil Son Khang ),이해방 ( 한국조직공학과 재생의학회 2006 조직공학과 재생의학 Vol.3 No.4
In order to fabricate tissue engineered biodisc, cells isolated from nucleus pulposus(NP) and annulus fibrosus(AF) have been characterized with sequential passages. NP and AF cells were separately by enzymatical digestion with 0.25wt% collagenase, respectively. Morphological changes were observed by phase contrast microscope and cell proliferation was counted by hemacytometer. To analyze the biosynthesis of glycosaminoglycan and gene expression of Type I and Type II collagen, safranin-O staining and RT-PCR have been carried out, respectively. Proliferation of NP and AF cells increased up to passage 3 and passage 6. Morphology of NP cells was maintained to passage 5 and it of AF cells was maintained to passage 4. In the result of safranin-O staining, NP and AF cells was stained positively. Type II collagen gene of NP cells was not expressed after passage 6 and expression of Type I collagen gene of AF cells was maintained up to passage 9. It can be expected that these results provide the important information for the application of tissue engineered biodisc.