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비타민 C를 함유한 락타이드 글리콜라이드 공중합체 지지체에서의 항염증, 항산화 효과
장나금(Na Keum Jang),이선의(Seon Eui Lee),차세롬(Se Rom Cha),김창현(Chang Hyun Kim),송정은(Jeong Eun Song),권순용(Soon Yong Kwon),강길선(Gilson Khang) 한국고분자학회 2016 폴리머 Vol.40 No.1
Poly(lactide-co-glycolide) (PLGA)는 대표적인 생분해성, 생체적합성을 가진 고분자로 조직공학 및 약물전달체로 널리 적용되고 있다. 그러나 이식 후에 PLGA는 에스테르결합으로 인해 가수분해가 일어나 분해산물의 영향으로 pH가 낮아져 조직에 염증반응을 유발한다. 비타민 C는 보통 식품, 약품에서 사용되고 있고, 강력한 항산화와 항염증 효과를 나타내는 것으로 알려져 있다. 그래서 조직공학 전달체로서의 PLGA와 비타민 C를 연구하였다. 본 연구에서는, 비타민 C가 포함된 PLGA 지지체를 제작하여 비타민 C의 생체 외 염증반응을 평가하였다. 지지체는 PLGA 무게당 3, 5, 10 wt%의 비타민 C를 첨가하여 캐스팅/염 추출법으로 3차원 비타민 C/PLGA 지지체로 제작하였다. 지지체의 물리적 특성은 압축강도와 SEM을 통해 조사하였다. 세포의 부착과 증식률은 MTT와 SEM 분석을 통해 실험하였다. 또한 RT-PCR과 FACS를 통해 항염증, 항산화 효과를 확인하였다. 결과는 공통적으로 5 wt% 비타민 C/PLGA 지지체가 항산화, 항염증 효과, 세포의 증식률의 면에서 다른 지지체보다 우수한 것으로 나타났다. 따라서 항염증 효과를 기초로하여, 5 wt% 비타민 C/PLGA 지지체는 미래의 조직공학에 쓰일 생체재료로 적용될 수 있을 것으로 사료된다. Typical biodegradable polymer poly(lactide-co-glycolide) (PLGA) has been widely applied to tissue engineering scaffolds and drug delivery systems. However, after the implantation, the ester bond of PLGA gets hydrolyzed and the resulting degradation products decrease the pH in the surrounding tissue causing local inflammatory reaction. Vitamin C is commonly used in food, pharmaceutical products and is known to exert potent antioxidant and anti-inflammatory activities. Thus, vitamin C is used with PLGA as carrier in tissue engineering. In this study, vitamin C incorporated PLGA scaffolds were fabricated and further the effects of vitamin C were evaluated on the inflammatory responses in vitro. The three-dimensional PLGA scaffolds were fabricated by adding of 3, 5, and 10 wt% vitamin C via a casting extraction method. Then the samples were examined; the mechanical properties of compressive strength and SEM. The cell attachment and proliferation were also studied the MTT, SEM, anti-inflammatory and antioxidant effects by RT-PCR and FACS. Results showed that 5 wt% vitamin C/PLGA scaffold was better than other scaffolds in terms of anti-inflammatory, antioxidant, cell attachment and proliferation. Based on the low level of inflammatory responses, the 5 wt% vitamin C/PLGA scaffold can be envisioned as a promising candidate for future biomedical applications.
헤스페리딘을 함유한 락타이드 글리콜라이드 공중합체 지지체에서 섬유륜 세포의 증식과 거동 평가
장나금(Na Keum Jang),고현아(Hyun Ah Ko),차세롬(Se Rom Cha),이선의(Seon Eui Lee),정현기(Hyun Ki Jeong),김수영(Su Young Kim),신재훈(Jae Hun Shin),김은영(Eun Young Kim),송정은(Jeong Eun Song),강길선(Gilson Khang) 한국고분자학회 2015 폴리머 Vol.39 No.5
항산화와 항염증으로 잘 알려진 헤스페리딘은 디스크 재생을 위해 효과적인 지지체로 제작하기 위해 사용하였다. 지지체는 poly(lactide-co-glycolic acid) (PLGA)에 헤스페리딘(0, 3, 5, 10%)을 첨가하여 염 침출법으로 제작하였고, 섬유륜 세포(AF)를 파종한 후 세포의 변화를 연구하였다. SEM, WST 그리고 RT-PCR 분석을 통해 세포의 부착과 증식, 세포의 표현형 분석을 실시하였다. 또한 면역조직화학염색을 통하여 AF세포의 생체 내 거동을 확인하였다. 그 결과, 5% 헤스페리딘이 함유된 PLGA 지지체가 가장 좋은 세포의 형태와 생체 적합성을 보여, 디스크 재생을 위한 지지체로서의 활용 가능성을 확인하였다. Hesperidin, a well-known antioxidant and anti-inflammatory agent, was smartly used for designing efficient scaffolds for disc regeneration. The scaffolds were synthesized via a salt-leaching method by adding hesperidin (0, 3, 5, & 10%) to poly(lactide-co-glycolic acid) (PLGA) followed by a cellular behaviors study by seeding annulus fibrosus (AF) cells. The cellular adhesion, proliferation and phenotypes were further evaluated by SEM, WST and RT-PCR, respectively. Additionally, AF cell behavior was studied by immunohisto-chemical staining in vivo. Results showed that PLGA scaffold with 5% hesperidin are best suited for disc regeneration, showing good cellular morphology and biocompatibility.
차세롬,조선아,이선의,장나금,조성준,송정은,강길선 한국고분자학회 2016 Macromolecular Research Vol.24 No.5
In this study, porcine small intestinal submucosa (SIS) without immune response were used to make SIS/ PLGA scaffolds with different SIS content i.e. 0, 10, 20, 40, and 80 wt%. Further, the attachment and proliferation of costal cartilage cells (CCs) in SIS/PLGA scaffolds were characterized. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to verify the effects of SIS on extracellular matrix (ECM) formulation. The CCs specific gene expression was confirmed by polymerase chain reaction (PCR). In conclusion, the sGAG production was formed to be higher in the 20 wt% SIS/PLGA scaffolds, which showed enhanced CCs cell growth and proliferation compared with other scaffolds.
한갑수,강길선,고현아,장나금,송정은 한국고분자학회 2014 Macromolecular Research Vol.22 No.12
The aim of the study was to investigate the effects of small intestinal submucosa (SIS) sponges, a naturalbiodegradable polymer scaffold, on the adhesion behavior and the proliferation of Schwann cells (SCs). SIS spongeswere prepared by varying the contents of SIS powders (1%, 2%, and 3% SIS concentration) and SCs were seededon the sponges and incubated to investigate the adhesion and proliferation of SCs. A series of analytical process wasconducted to observe the water absorption rate of sponges and confirmed the proliferation and morphology of SCsby scanning electron microscopy (SEM), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT)assay, reverse transcription polymerase chain reaction (RT-PCR), and histological evaluation. The best adhesionbehavior and the proliferation of SCs were observed on the sponge with 2% powder content. Also, analysis of messengerribonucleic acid (mRNA) expression using the neuron-specific enolase (NSE) and neurofilament protein (NF)nerve markers showed that SC gene expression was excellent in the 2% SIS sponge. In vivo hematoxylin and eosin(H&E) staining and immunofluorescence results showed that cells and tissue formed well in the pores of spongeswith 2% and 3% SIS powder contents, compared with formation in the 1% SIS sponge. The SIS sponges provided anunsuitable habitat for the adhesion and proliferation of cells in that the pore size was small. In this study, the adhesionbehavior and the proliferation could be changed depending on the pore size and SIS powder content in the sponges. Thus, an appropriate powder content of SIS should be used to prepare SIS sponge scaffolds.
탈미네랄화된 골분 스펀지에서 쥐지방줄기세포의 골분화 유도
김창현(Chang Hyun Kim),장나금(Na Keum Jang),김도경(Do Kyung Kim),송야성(Yah Seong Song),송정은(Jeong Eun Song),강길선(Gilson Khang) 한국고분자학회 2017 폴리머 Vol.41 No.1
탈미네랄화된 골분(DBP)은 콜라겐, 칼슘 그리고 프로테오글리칸을 포함한다. DBP는 조직공학 분야에서 널리 사용되는 생체재료로, 새로운 뼈나 연골 성장의 기초를 다지는데 도움을 주는 생체 활성 분자를 포함한다. 본 연구에서는 천연 생체재료인 DBP를 1, 2, 3% 비율의 스펀지형태로 제작하였다. 또한 체내 지방에서 쉽게 얻을 수 있는 쥐지방줄기세포(rASCs)를 DBP 스펀지에 파종한 후, 골분화를 확인하기 위해 다음과 같은 연구를 실시하였다. 먼저, 세포의 부착과 증식의 효과를 확인하고자 SEM과 MTT분석을 실시하였으며, 골분화의 발현 정도와 잠재력 확인을 위해 ALP 활성과 RT-PCR을 실시하였다. 조직화학적 염색은 쥐에 DBP 스펀지를 이식한 후 1주, 4주 및 8주 후에 적출하여, Alizarine Red S 염색과 Von Kossa 염색을 통해, 생체 내에서 골분화의 효과를 확인하였다. 그 결과, 1% DBP 스펀지가 rASCs의 골분화능을 증가시키는데 적합한 환경을 제공한다는 것을 확인할 수 있었다. Demineralized bone particle (DBP) is a well-studied biomaterial, composed of collagen, calcium and proteoglycan. It is widely used in tissue engineering and regenerative medicine, because it contains powerful bioactive molecules that facilitate new bone or cartilage growth. Herein, we fabricated 1, 2 and 3% sponges using natural biomaterial DBP and studied its efficiency in rat adipose stem cells (rASCs) as models. After rASCs were seeded on DBP sponges, we evaluated rASCs adhesion, proliferation, cell attachment, proliferation and osteogenesis using SEM, MTT, and bone differentiation expose using ALP assay, RT-PCR in vitro. Histological examinations were performed at 1, 4 and 8 weeks after implantation DBP sponge to assess the effect of bone differentiation in vivo using Alizarin Red S, Von Kossa staining. We found that 1% DBP sponge can provide suitable environment for bone differentiation of rASCs.