• 폴리머
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• 제32권5호
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• pp.403-408
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• 2008

케라틴은 울, 머리카락, 손톱 등을 형성하는 섬유단백질의 주요성분으로 유용한 생체재료이다. 골수간염 줄기세포를 이용한 조직공학 적용을 위해 Poly(4-lactide-co-glycolide) (PLGA)에 함량별로 케라틴을 함유한 지지체를 용매 캐스팅/염 추출법을 이용하여 제조하였다. 제조된 지지체의 표면과 단면의 형태를 전자현미경(SEM)으로 관찰하고 특성분석을 위해 다공도, 표면 적심성, 물 흡수성, 그리고 열적성질을 분석하였다. 이 후 쥐에서 분리한 골수간엽줄기세포를 지지체에 파종하여 세포의 증식율을 (4,5-dimethylthiazol-2-yl)-2.5-diphenyl-tetrazolium bromide(MTT) 분석방법을 이용하여 측정하였다. 천연/합성 하이브리드 담체인 케라틴/PLGA 지지체는 PLGA 단독으로 제조된 지지체와 비교 시 골수간엽줄기세포의 생장에 유익한 환경을 제공함을 확인하였다.

• 폴리머
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• 제33권4호
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• pp.353-357
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• 2009

분무건조는 약물의 입자 제조에 널리 사용되고 있는 효과적이고 안정적인 공정 중 하나이다. 과거 분무건조의 방법에서는 입자의 응집을 조절하기 쉽지 않았으며, 크기 조절에도 많은 어려움을 겪어 왔다. 특히 고분자 입자의 제조는 저분자 유기물질인 식품, 의약원료 등과 비교하여 상대적으로 제조하기 어려운 면이 있었다. 본 연구에서는 기존의 분무건조기를 개조하여 노즐에 전기장을 외부에서 가해 줌과 동시에 동축 이중 노즐을 사용하여 고분자 입자의 제조를 시도하였다. 내부 고분자 물질로는 폴리에틸렌글리콜과 폴리락타이드코글리코라이드를, 외부 물질로는 락토오스를 사용하였다. 그 결과 전기장을 사용하지 않는 분무건조에 비해 비교적 일정한 크기와 모양을 제조할 수 있었으며, 입자 간의 응집을 줄일 수 있음을 확인하였다. 전기분무건조된 PLGA 분말은 주로 $2{\sim}5{\mu}m$ 기인 둥근 모양의 입자로 구성되었다.

• Macromolecular Research
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• 제10권3호
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• pp.158-167
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• 2002

In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

• Journal of Pharmaceutical Investigation
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• 제34권5호
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• pp.369-377
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• 2004

The objective of this study was to investigate the patterns of protein leaching to an external phase during an ethyl acetate-based, double emulsion microencapsulation process. An aqueous protein solution (lactoglobulin, lysozyme, or ribonuclease; $W_1$) was emulsified in ethyl acetate containing poly-d,l-lactide-co-glycolide 75:25. The $W_1/O$ emulsion was transferred to a 0.5% polyvinyl alcohol solution saturated with ethyl acetate $(W_2)$. After the double emulsion was stirred for 5, 15, 30, or 45 min, additional 0.5% polyvinyl alcohol $(W_3)$ was quickly added into the emulsion. This so-called quenching step helped convert emulsion microdroplets into microspheres. After 2-hr stirring, microspheres were collected and dried. The degree of protein leaching to $W_2$ and/or $W_3$ phase was monitored during the microencapsulation process. In a separate, comparative experiment, the profile of protein leaching to an external phase was investigated during the conventional methylene chloride-based microencapsulation process. When ethyl acetate was used as a dispersed solvent, proteins continued diffusing to the $W_2$ phase, as stirring went on. Therefore, the timing of ethyl acetate quenching played an important role in determining the degree of protein microencapsulation efficiency. For example, when quenching was peformed after 5-min stirring of the primary $W_1/O$ emulsion, the encapsulation efficiencies of lactoglobulin and ribonuclease were $55.1{\pm}4.2\;and\;45.3{\pm}7.6%$, respectively. In contrast, when quenching was carried out in 45 min, their respective encapsulation efficiencies were $39.6{\pm}3.2\;and\;29.9{\pm}11.2%$. By sharp contrast, different results were attained with the methylene-chloride based process: up to 2 hr-stirring of the primary and double emulsions, less than 5% of a protein appeared in $W_2$. Afterwards, it started to partition from $W_1\;to\;W_2/W_3$, and such a tendency was affected by the amount of PLGA75:25 used to make microspheres. Different solvent properties (e.g., water miscibility) and their effect on microsphere hardening were to be held answerable for such marked differences observed with the two microencapsulation processes.

• Bulletin of the Korean Chemical Society
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• 제35권8호
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• pp.2342-2348
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• 2014

Poly(lactic-co-glycolic acid) (PLGA) were grafted to both ends of Pluronic$^{(R)}$ F68 ($(EO)_{75}(PO)_{30}(EO)_{75}$) triblock copolymer to produce poly{(lactic acid)$_m$-co-(glycolic acid)$_n$}-b-poly(ethylene oxide)$_{75}$-b-poly(propylene oxide)$_{30}$-b-poly(ethylene oxide)$_{75}$-b-poly{(lactic acid)$_m$-co-(glycolic acid)$_n$} (PLGA-F68-PLGA) pentablock copolymers. Molecular weights of PLGA blocks were controlled and five kinds of pentablock copolymers with different PLGA block lengths were synthesized using in-situ ring-opening polymerization of D,L-lactide and glycolide with tin(II) 2-ethylhexanoate ($Sn(Oct)_2$) catalyst. PLGA-F68-PLGA pentablock copolymers were characterized by $^1H$- and $^{13}C$-NMR, GPC, and TGA. The numbers (2m, 2n) of repeating units for lactic acid and glycolic acid inside PLGA segments were obtained as (48, 17), (90, 23), (125, 40), (180, 59), and (246, 64), with $^1H$-NMR measurement. From NMR data, the resultant molecular weights were determined in the range of 12,700-29,700, which were similar to those obtained from GPC. Polydispersity index was increased in the range of 1.32-1.91 as the content of PLGA blocks increased. TG and DTG thermograms showed discrete degradation traces for PLGA and F68 blocks, which indicate the weight fractions of PLGA blocks in pentablock copolymers can be calculated by TG profile and it is possible to remove PLGA block selectively. Hydrodynamic radius and radius of gyration of pentablock copolymer micelle were obtained in the range of 46-68 nm and 31-49 nm, respectively, in very dilute (i.e. 0.005 wt %) aqueous solution of THF:$H_2O$ = 10:90 by volume at $25^{\circ}C$.

• Molecules and Cells
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• 제38권6호
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• pp.518-527
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• 2015

Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and c ell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as $p21^{WAF}$, cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.141-144
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• 2000

Highly open porous polymer matrices are required for high density cell seeding, efficient nutrient, and oxygen supply to the cells cultured in the three dimensional matrices. However, there are severe problems of mass transfer limitations within the cell/scaffolds culture system. Thus we hypothesize that continuos-flow culture conditioning of cells with the scaffolds may improve the cell viability and the differentiated function. In this study, we fabricated porous PLGA scaffolds by using gas-foaming/salt-leaching method as previous described. Viscous PLGA gel paste contains ammonium bicarbonate particulates, acting as a gas-foaming agent as well as a salt-leaching porogen, were cast into Teflon mold and dried. Ammonium bicarbonate salt upon contact to an acidic aqueous solution evloves gaseous ammonia and carbon dioxide by itself. And we conjugated galactose moiety [AGA; $N-(aminobuty1)-O-{\beta}-D-galactopyranosyl-(1{\rightarrow}4)-D-glucoamide]$ to the terminal end group of a PLGA to increase the cell adhesion and matain the differentiated function of hepatocytes. Cell-seeded scaffolds were secured in a flow bioreactor chamber and exposed to continuous flow at 5 ml/min. As a result of our study, the high yield of hepatocytes attachment was accomplished by increasing the concentration of PLGA-AGA conjugate in polymer scaffolds and cells in the scaffolds under continuos flow condition maintained a high level of viability and albumin secretion rate of cultured hepatocytes showed a higher level that of control groups.

• KSBB Journal
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• 제23권5호
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• pp.408-412
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• 2008

본 연구에서는 당뇨병치료제인 로시글리타존약물를 생분해성 PLGA 나노입자에 봉입시킴으로서 위장흡수율과 물에 대한 용해도를 증가시키기 위한 나노제제 개발에 기반을 두고 있다. 특히 제조조건에 따라 형태 및 크기가 조절가능한 나노입자를 제조하고자 하였고, 실험결과 Emulsion-evaporation방법을 사용하여 100-150 nm 크기의 고른 입자분포를 가진 나노입자를 제조하였다. 다양한 농도의 약물 존재하에서 나노입자를 제조함으로서 1%까지의 약물이 80% 이상의 봉입율로 제조될 수 있음을 확인하였다. 또한 나노입자의 크기는 PVA양을 조절하면서 크기분포를 제어하였다. 36시간 동안의 용출실험 결과 초기 약간의 Burst effect가 있었으나 36시간동안 일정하게 약물이 용출되어 나옴을 확인하였다. 앞으로의 연구를 통해, 고효율의 경구제제용 당뇨병치료제 운반체 개발에 중점을 둘 예정이다.

• Journal of Pharmaceutical Investigation
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• 제34권1호
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• pp.1-14
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• 2004

A promising approach to the development of a new single-step vaccine, which would eliminate the requirement for multiple injections, involves the encapsulation of antigens into microspheres. Biodegradable poly(lactide-co-glycolide) (PLGA) microspheres gave us a bright insight for controling antigen release in a pulsatile fashion, thereby mimicking two or tree boosting injections. However, in spite of the above merits, the level of immunization induced by a single-shot vaccination is often lower tan two doses of alum-adsorbed antigen. Therefore, optima modification of the microsphere is essential for the development of single-step vaccines. In the review, we discuss the stability of antigen in microsphere, safety and non-toxic in human and encapsulation technology. Also, we attempted to outline relevant physicochemical properties on the immunogenicity of microsphere vaccine and attainment of pulsatile release pater by combination of different microsphere, as well as to analyze immunological data associated with antigen delivery by microsphere. Although a lot of variables are related to the optimized microsphere formulation, we could conclude that judicious choice of proper polymer type, adjustment of particles size, and appropriate immunization protocol along with a suitable adjuvant might be a crucial factor for the generation of long-lasting immune response from a single-step vaccine formulation employing PLGA microsphere.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.421-424
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• 2000

Cartilage injuries are frequent nowadays. The previous surgical treatment of cartilage defect was limited. Another approach in the treatment of cartilage injuries is the use of reconstitute cartilage consisting of chondrocytes cultured in suitable biodegradable scaffolds. Current studies have demonstrated the compatibility of chondrocytes with different biomaterials and the chondrogenesis in various types of porous scaffolds. The cell ingrowth into the porous scaffolds is modulated by initial cell loading efficiency. Therefore, well-interconnected pore structure and even pore distribution of the scaffolds are essential for efficient cell seeding. According to our previous work, well-interconnected macroporous scaffolds can be prepared by gas-foaming/salt-leaching method using ammonium bicarbonate salt as porogen additives. In this work, primary chondrocytes were cultured in PLGA 65/35 scaffolds fabricated by using our method. Cells seeded in the scaffolds showed well distribution by agitated seeding method. Histochemical staining of proteoglycans present in the scaffolds was used to visualize the chondrocyte ingrowth in the scaffolds. At 3 weeks, the population of chondrocytes was increased for the most part of the scaffolds, and extra cellular matrix (ECM) secretion was increased as culture periods progressed.