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PLGA미립구의 크기에 따른 인간디스크세포의 증식 및 표현형 분석
소정원 ( Jeong Won So ),김순희 ( Soon Hee Kim ),백미옥 ( Mi Ock Baek ),임지예 ( Ji Ye Lim ),노혜원 ( Hye Won Roh ),이나리 ( Na Ri Lee ),김문석 ( Moon Suk Kim ),유규하 ( Gyu Ha Ryu ),조양하 ( Yang Ha Cho ),이승진 ( Seung Jin Lee ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
The PLGA microspheres can create an environment conductive to promising candidate as an injectable system for tissue engineered intervertebral disc(IVD). The final goal of this study was to investigate the condition of PLGA microspheres as microcarriers for IVD cells. We prepared PLGA microspheres using oil-in-water(O/W) solvent evaporation method and investigated the size variation of the microspheres(180~425 μm). Human disc cells were used in a prospective study of attachment, proliferation, and gene expression within various size of PLGA microspheres. Mophology of cell attached on the microspheres was studied using scanning electron microscopy (SEM). Cellular behavior in response to various size of PLGA microspheres was analyzed using MTT assay and reverse transcriptase - polymerase chain reaction(RT-PCR). SEM showed that cells could be bridge between microspheres with elongated shapes. MTT assay revealed that the cells could be attach, proliferate and spread on PLGA microspheres, in particular on the ones having size below 250 μm. In RT-PCR results, we observed strongly specific gene expression of type collagen at more 250 μm microsphere. In conclusion, this result will be used to injectable system for disc regeneration.
PLGA 담체의 제조방법에 따른 골수간엽줄기세포의 골분화 유도
소정원 ( Jeong Won So ),김순희 ( Soon Hee Kim ),백미옥 ( Mi Ock Baek ),임지예 ( Ji Ye Lim ),노혜원 ( Hye Won Roh ),이나리 ( Na Ri Lee ),유규하 ( Gyu Ha Ryu ),조양하 ( Yang Ha Cho ),이승진 ( Seung Jin Lee ),강길선 ( Gilson Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.1
A key component in tissue engineering for bone regeneration is the scaffold that serves as a template for cell interaction and the formation of bone-extracellular matrix to provide structural support to the newly formed tissue. In this study, poly(L-Lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/salt-leaching, ice particle-leaching and gas-forming/salt-leaching. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole- 2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed ALP and osteocalcin m RNA expression. Scaffold with BMSCs were implanted into the back of athymic nude mouse to observe in vivo behavior. Thin section were cut from paraffin embedded tissues and histological sections were stained by trichrome, immunohistochemical, von kossa staining. The cell proliferation and ALP activity of BMSCs in the scafoolds was the highest in the scaffolds using solvent casting/salt-leaching method. In RT-PCR results, in the all type of scaffolds observed specific gene expression of ALP, osteocalcin. By histological staining, we could observed collagen and calcification region in scaffolds by solvent casting/salt leaching method. Thus Solvent casting/salt-leaching method provided highly porous structure with good interconnections between each pores, which can support the surface of cell proliferation, differentiation. This result indicates that solvent casting/salt-leaching is useful for osteogenesis of BMSCs.
탈미네랄화된 골분/히알루론산을 이용한 조직공학적 지지체의 제조 및 특성분석
임지예 ( Ji Ye Lim ),김순희 ( Soon Hee Kim ),소정원 ( Jeong Won So ),이종문 ( John M. Rhee ),강길선 ( Gil Son Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.3
Demineralized bone particle(DBP) that affects to cell proliferation and differentiation has been used to clinical trial as natural material. hyalunonic acid(HA) called natural moisturizer can be used as biomaterials. In this study, we tried to design and characterize novel DBP-HA sponge. DBP-HA sponge were prepared by freeze-drying method after addition of 1 wt% HA solution into fabricated DBP sponge. 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-HA sponge. Cellular attachment was assayed by 3-(4,5-dimethylthiazole-2-yl)- 2,5-diphenyltertazolium-bromide(MTT) test. DBP-HA sponges were characterized by scanning electron microscopy, fourier transeform infrared spectrometer and water ubsorption ability. We demonstrated presence of HA in DBP-HA sponge from C-O functional group observed by the FT-IR analysis. Moreover, we confirmed high water uptake ability and high cell viability in DBP-HA sponge. Therefore, we could expect that DBP-HA scaffolds are applicable for the tissue regeneration.
SIS/PLGA 필름에서 SIS 함량이 디스크세포의 증식에 미치는 영향
임지예 ( Ji Ye Lim ),김순희 ( Soon Hee Kim ),소정원 ( Jeong Won So ),최진희 ( Jin Hee Choi ),이종문 ( John M. Rhee ),신형식 ( Hyung Shik Shin ),강길선 ( Gil Son Khang ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.3
Porcine small intestine submucosa is accellular tissue and widely used as a biomaterials without immunorejection responses. Poly(lactide-co-glycilide) (PLGA) has controllable biodegradability and good biocompatibility. In this study, we evaluated 2-dimensional porcine small intestine submucosa (SIS)/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. We prepared natural/synthetic hybrid films using 0, 10, 20, 40 and 80 wt% of SIS by the solvent evaporation method. SIS/PLGA films were characterized by scanning election microscopy (SEM). Disc cells were seeded on PLGA and SIS/PLGA film and we confirmed the effects of adhesion and proliferation on disc cells according to content of SIS by cell count and SEM observation. Contents of 10 wt% SIS positively affected to proliferate and maintain of disc phenotype in SIS/PLGA Film.
PLGA/DBP 하이브리드지지체와 섬유륜세포 및 수핵세포를 이용한 바이오디스크의 재생
임지예 ( Ji Ye Lim ),김순희 ( Soon Hee Kim ),박상욱 ( Sang Wook Park ),소정원 ( Jeong Won So ),백미옥 ( Mi Ock Back ),김문석 ( Moon Suk Kim ),강길선 ( Gilson Khang ),이종문 ( John M. Rhee ),이해방 ( Hai Bang Lee ) 한국조직공학과 재생의학회 2008 조직공학과 재생의학 Vol.5 No.1
We fabricated poly(lactide-co-glycolide)(PLGA) scaffolds impregnated 0, 10, 20, 40, and 80 wt% demieneralized bone particle(DBP)(PLGA/DBP) to investigate the effect of cell viability and proliferation of intervertebral disc(IVD) cells. PLGA is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. DBP contains bioactive molecules. Annulus fibrosus(AF) and nucleus pulposus (NP) cells were seeded in PLGA and PLGA/DBP scaffold to evaluate mRNA expression and a proliferation of AF and NP cells according to content of DBP. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole- 2-l)-2,5-diphenyltetrazolium-bromide(MTT) test. Reverse transcription polymerase chain reaction(RT-PCR) was assessed to measure mRNA expression for Type I collagen and Type II collagen of IVD cells in scaffolds. Scaffolds were implanted in subcutaneous of athymic nude mouse for histology in vivo. The formation of disc-like tissue appeared in PLGA/DBP scaffold than control. Especially, 40 and 80 wt% of DBP positively affected to proliferation and maintenance of phenotype in scaffold. These information would contribute to design of PLGA/DBP scaffold for tissue engineered IVD regeneration.
소정원,이광희,라지영,안소연,김윤희,반재혁 大韓小兒齒科學會 2010 大韓小兒齒科學會誌 Vol.37 No.1
Ectopic eruption is caused by an abnormal direction of eruptive path, most common in maxillary first molar, mandibular lateral incisor, and maxillary canine, and sometimes mandibular first molar. Ectopic eruption of first molar leads to abnormal root resorption of second deciduous molar, which, if left untreated, could cause premature loss of second deciduous molar; mesial tilting and rotation of first permanent molar; lack of space for eruption of second premolar; and occlusal problems. Therefore early treatment is advised when diagnosed as ectopic eruption. Treatment of ectopic eruption in the first permanent molar involves providing proper guidance for the direction of eruption using interproximal wedging and distal tipping methods while preserving second deciduous molar. This case report shows satisfactory results of the ectopic eruption of mandibular first molars in young patients who were treated with Humphrey appliance and Halterman appliance. 이소맹출은 치아가 비정상적인 위치로 맹출하는 경우를 말하며, 주로 상악 제1대구치, 하악 측절치, 상악 견치에서 발생하며 하악 제1대구치에서는 드물게 발생한다. 제1대구치의 이소맹출은 흔히 제2유구치의 비정상적인 치근흡수를 야기하며, 이를 방치하게 되면 제2유구치의 조기상실, 제1대구치의 근심경사 및 회전, 제2소구치의 맹출 공간 부족 및 교합문제 등을 야기하게 되므로 이소맹출로 진단될 경우 조기 치료가 추천된다. 이소맹출은 대개 통상적인 방사선검사를 통해 발견되나, 간혹 제2유구치의 치근흡수가 심할 경우 치수가 감염되어 동통을 야기하는 경우도 있다. 이소맹출의 치료는 제2유구치를 보존하면서 제1대구치의 맹출 방향을 바로 잡아 주는 것으로 크게 치간이개(interproximal wedging)와 원심경사이동(distal tipping)을 이용한 방법을 사용한다. 본 증례들은 하악 제1대구치의 이소맹출을 보이는 환아들로, Humphrey appliance와 Halterman appliance를 이용하여 양호한 치료 결과를 얻었기에 보고하는 바이다.