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Osteogenic Differentiation of Bone Marrow Stem Cells Using Thermo-Sensitive Hydrogels (온도감응성 수화젤을 이용한 골수간엽줄기세포의 골분화 유도)

  • Kim, Sun-Kyung;Hyun, Hoon;Kim, Soon-Hee;Yoon, Sun-Jung;Kim, Moon-Suk;Rhee, John-M.;Khang, Gil-Son;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.30 no.3
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    • pp.196-201
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    • 2006
  • Poly (ethylene glycol)-based diblock and triblock thermo- sensitive polyester copolymers were investigated for application on tissue engineering and injectable biomaterials in drug delivery system due to their nontoxicity, biocompatibility and biodegradability. We synthesized the diblock copolymers consisting of methoxy poly (ethylene glycol) (MPEG) (Mn=750 g/mole) and poly $(\varepsilon-caprolactone)$ (PCL) by ring opening polymerization of $\varepsilon-CL$ with MPEG as an initiator in the presence of HCl $Et_2O$. The effect of diblock copolymers on in vivo osteogenic differentiation of rat bone marrow stromal cells (BMSCS) with and without the presence of osteogenic supplements (dexamethasone) was investigated. Thin sections were cut from paraffin embedded tissues and histological sections were stained by H&E, von Kossa, and immunohistochemical staining for osteocalcin. In conclusion, dexamethasone containing thermo- sensitive hydrogel might be improved osteogenic differentiation of BMSCs. We expect the osteoinduction effect to be excellent when it uses stem cell or other osteogenic materials.

Preparation and Characterization of Hyaluronic Acid Loaded PLGA Scaffold by Emulsion Freeze-Drying Method (히알루론산을 함유한 PLGA 지지체의 제조 및 특성결정)

  • Ko, Youn-Kyung;Kim, Soon-Hee;Jeong, Jae-Soo;Park, Jung-Soo;Lim, Ji-Ye;Kim, Moon-Suk;Lee, Hae-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.31 no.6
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    • pp.505-511
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    • 2007
  • Poly(lactide-co-glycolide)(PLGA) and hyaluronic acid (HA) has been widely used as biocompatible scaffold materials to regenerate tissue. In this present study, we fabricated microporous PLGA and HA loaded PLGA scaffolds by a emusion freeze-drying method. In order to confirm that the release profile of cytokine or water-soluble drugs, we manufactured the granulocyte macrophage colony stimulating factor(GM-CSF) loaded PLGA and HA-PLGA scaffold. All scaffolds were characterized using scanning electron microscope(SEM), mercury porosimeter and wettability measurement. Cell proliferation and viability were assessed by a 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) test. The porosity of HA-PLGA scaffold was greater than 95% with the total pore area of $261\;m^2/g$. The HA-FLGA scaffold exhibited well interconnected pores to allow greater cell adhesion and prolixferation. It was proven by higher cell viability in the HA-PLGA scaffold than PLGA alone. This may be due to the enhanced natural properties and higher water retention capacity of HA.

Effects of Laminated Cylindrical Scaffolds of Keratin/Poly(lactic-co-glycolic acid) Hybrid Film on Annulus Fibrous Tissue Regeneration (케라틴/PLGA 복합체 필름의 적층 원통형 지지체가 섬유륜 재생에 미치는 영향)

  • Lee, Seon-Kyoung;Hong, Hee-Kyung;Kim, Su-Jin;Kim, Yong-Ki;Lee, Dong-Won;Khang, Gil-Son
    • Polymer(Korea)
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    • v.34 no.5
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    • pp.474-479
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    • 2010
  • We developed laminated cylindrical scaffolds composed of poly(lactide-co-glycolide)(PLGA) and keratin, and investigated their potential for tissue engineering and disk regeneration. The scaffold was designed to have two parts, i.e. inner cylinder and outer disk, to mimic a natural disk. The outer disk was composed of PLGA and the inner cylinder was prepared using PLGA film or PLGA/keratin hybrid film. In this study, we investigated the effects of keratin on the growth and proliferation of annulus fibrous(AF) cells in the cylindrical scaffolds. Scaffolds containing PLGA/keratin films showed a significantly higher cell proliferation and expression of collagen I and II than the counterpart with PLGA films. Keratin containing scaffolds also exhibited an excellent mechanical strength, demonstrating that keratin influences the proliferation of annulus fibrous cells. The results provide valuable information on PLGA/keratin films for tissue engineered disk regeneration.

Effect of PLGA/Silk Fibroin Hybrid Film on Attachment and Proliferation of Schwann Cells (실크피브로인을 함유한 PLGA 하이브리드 필름이 슈반세포의 부착과 증식에 미치는 영향)

  • Kim, Hye-Lin;Yoo, Han-Na;Park, Hyun-Jin;Kim, Yong-Gi;Lee, Dong-Won;Kang, Young-Sun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.1
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    • pp.7-12
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    • 2011
  • Poly(lactic-co-glycolic acid) (PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. Also, it can be easily fabricated into many shapes. Silk fibroin contains powerful bioactive molecules. We fabricated natural/synthetic hybrid films using 0, 10, 20, 40 and 80 wt% of silk fibroin. Schwann cells (SCs) were seeded on PLGA/silk fibroin hybrid films and confirmed the effects of adhesion and proliferation on SCs according to the content of silk fibroin. In this study, we confirmed PLGA/silk fibroin hybrid film containing 40% and 80% of silk fibroin interrupted adhesion and proliferation of SCs. Films containing 10% and 20% of silk, however, provided suitable environment for growth and proliferation of SCs. These results suggest that silk fibroin provides suitables surface to neural cells and its proper content provides proper culture conditions to improve cell adhesion and proliferation.

The Effect of Transplantation of Schwann Cell and SIS Sponge on the Injured Peripheral Nerve Regeneration (슈반세포와 SIS 스폰지의 이식이 손상된 말초 신경 재생에 미치는 영향)

  • Kim, Cho-Min;Kim, Soon-Hee;Kim, Su-Mi;Park, Sang-Wook;Lee, Il-Woo;Kim, Moon-Suk;Rhee, John-M.;Khang, Gil-Son;Lee, Hai-Bang
    • Polymer(Korea)
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    • v.32 no.1
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    • pp.49-55
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    • 2008
  • It is recognized that Schwann cells (SC) are essential for peripheral nerve development and regeneration. SIS (small intestinal submucosa) consists of some growth factors which can stimulate cell activity without immune rejection responges. SCs were harvested from the femurs and tibias of female Fischer rat and then suspended with $2{\times}10^6$ cell/sponge in SIS sponge. Fischer rat received an implant consisting of the SCs and the SIS sponge at the place of a 5 mm gap created by the sciatic nerve resection. Thin sections were stained with H &E staining and immunostaining of S-100, GFAP and NF after 1, 2, and 4 weeks. It was observed that the effects of the SIS sponge with SCs on neuroinduction(Group II, with scaffold & cell) are strong as much as uninjured model(Control I), and significantly stronger than SIS sponge model (Group 1, with scaffold only) and blank model (Control II). In conclusion, these results suggest that SIS sponge filled with SCs may have an important role for peripheral nerve regeneration of tissue engineering.

Effects of Demineralized Bone Particle Loaded Poly(lactic-co-glycolic acid) Scaffolds on the Attachment and Proliferation of Costal Cartilage Cells (탈미네랄화된 골분/PLGA 지지체에서 늑연골 세포의 부착과 성장에 미치는 영향)

  • Cho, Sun Ah;Song, Jeong Eun;Kim, Kyoung Hee;Ko, Hyun Ah;Lee, Dongwon;Kwon, Soon Yong;Chung, Jin Wha;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.5
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    • pp.632-637
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    • 2013
  • It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

Sustained Release of Proteins Using Small Intestinal Submucosa Modified PLGA Scaffold (SIS로 개질된 PLGA 담체에서의 단백질의 서방화)

  • Ko, Youn-Kyung;Choi, Myung-Kyu;Kim, Soon-Hee;Kim, Geun-Ah;Lee, Hai-Bang;Rhee, John-M.;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.3
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    • pp.199-205
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    • 2008
  • In this study, we fabricated poly (lactide-co-glycolide) (PLGA) scaffold modified with small intestinal submucosa (SIS) as a drug delivery matrix of bioactive molecules. SIS derived from the submucosa layer of porcine intestine has been widely used as biomaterial because of low immune response. PLGA scaffold was prepared by the method of solvent casting/salt leaching. Novel composite scaffolds of SIS/PLGA were manufactured by simple immersion method of PLGA scaffold in SIS solution under vacuum. SEM observation shows that PLGA and SIS/PLGA scaffolds have interconnective and open pores. Especially, SIS/PLGA scaffold showed that micro-sponge of SIS with interconnected pore structures were formed in the pores of PLGA scaffold. In order to assay release profile of proteins, we manufactured FITC conjugated BSA loaded PLGA and SIS/PLGA scaffold. And the release amount was identified by fluorescence intensity using the fluorescence spectrophotometer. The initial burst of BSA containing SIS/PLGA scaffolds was lower than that of PLGA scaffolds resulting in constant release. And release of BSA in SIS/PLGA scaffold was fast and incremental because of the increased content of BSA. In conclusion, we confirmed that penetrated SIS solution prevented the initial burst of BSA and PLGA modified with SIS scaffold is useful as protein carriers with controlled release pattern.

Composting Effectiveness of A Sundry System with A Bin-type Composter for Recyle of Animal Wastes (축분뇨처리를 위한 Bin형 부숙조- Sundry 시스템의 퇴비화효율 평가)

  • 최홍림;김현태;정영윤
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.35 no.2
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    • pp.92-103
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    • 1993
  • A sunday system with a horizontal bin-type composter was constructed and operated to evaluate its composting performance for four days for each test in October, 1992. A sundry system is one of popular systems for composting livestock manure, of which main benefit is to utilize unlimited, clean, and free solar radiation. A rectangular concrete bin(composter) with dimension of 300cm(length) X90cm(width) X60cm(height) was bedded alternatively with four lanes of aeration pipes and heating pipes, and was insulated at three walls with 50mm styrofoam. Each aeration pipe of a diameter of 25mm had 4mm perforated holes at every 15cm longitudinally, and supplied air of about 2m$^3$/min to the composter to maintain aerobic condition . A stirrer rotating at 1 rpm made one round trip every 20 minutes on the conveying chain along the the length of the composter. Five tests (Test 1~Test 5) were implemented to evaluate the composting effectiveness of a sundry system with a horizontal bin-type composter. Treatments of two levels of the mixture ratio of swine manure and paper sludge cakes(manure : paper sludge cakes= 1 : 4 and 1 : 2) and two levels of the water content(W/C ; 70% and 50%) were made to test the significance of the physicochemical properties for decomposition of the mixture materials. Temperature, C/N ratio, water content, microbial activity of the composting materials were taken measurements to evaluate its performance with the lapse of composting time for tests. A small-scale sundry system with a bin-type composter did not appear to be an appropriate system for composting livestock manure. Since heat generation by the composting materials could not overcome heat loss due to areation in a small-scale composter, a proper thermal enviroment could not be maintained to propagate massively thermopilic microorganism relatively in a short period of time. Different from the result of Chol et al.(1992) 6), a temperature variation of the composting materials did not show the peak clearly and C/N ratio didn't lower with time as expected. Mesophilic microoragnism seemed to play an important role for decomposition of the mixture materials. A sundry system with a bin-type composter may be good for a large-scale livestock farm household which may produce enough animal manure. Therefore a decision should be made very carefully to choose a system for composting livestock waste.

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pH Solubility Properties and Improved Dissolution of Pranlukast as an Poorly Water-soluble Model Drug Prepared by Spray-drying with Plasdone S-630 (플라스돈 S-630과 함께 분무건조된 모델 난용성 약물로서 프란루카스트의 pH 용해도 특성 및 용출률 개선)

  • Cho, Won-Hyung;Lee, Young-Hyun;Song, Byung-Joo;Yoo, Seok-Cheol;Lim, Dong-Kyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.4
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    • pp.277-283
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    • 2011
  • Solid dispersion is mainly used for improved dissolution of poorly water-soluble drugs. Solid dispersion of pranlukast was prepared by spray-drying with plasdone S-630. When pH of water was high, pranlukast was highly soluble in the solubility experiment of solid dispersions with varying pH. The particle size of pranlukast particles in solid dispersions was measured to be in nanometers scale based on particle size analysis. Zeta-potential analysis confirmed the negative charge of solid dispersion. SEM was used to observe the surface of solid dispersion, which confirmed spherical morphology, DSC and XRD confirmed the amorphous nature of solid dispersions. The in vitro test was carried out to find improved dissolution rate of pranlukast solid dispersion in simulated juice gastric and a controlled experiment was carried out to compare pranlukast solid dispersions with a conventional drug (Onon$^{(R)}$), These results showed the dissolution properties of pranlukast solid dispersions prepared by spray drying proper for the oral pharmaceutical formulation.

The Effect of Pore Sizes on Poly(L-lactide-co-glycolide) Scaffolds for Annulus Fibrosus Tissue Regeneration (조직공학적 섬유륜재생을 위한 PLGA 지지체 제조시 다공크기의 영향에 관한 연구)

  • So, Jeong-Won;Jang, Ji-Wook;Kim, Soon-Hee;Choi, Jin-Hee;Rhee, John-M.;Min, Byung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.6
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    • pp.516-522
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    • 2008
  • Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues and the ingrowth of tissue have been reported to be dependent directly of the porosity, pore diameter, pore shape, and porous structure of the scaffold. In this study, porous poly (L-lactide-co-glycolide) (PLGA) scaffolds with five different pore sizes were fabricated to investigate the effect of pore sizes for AF tissue regeneration. Cellular viability and proliferation were assayed by MTT test. Hydroxyproline/DNA content of AF cells on each scaffold was measured. sGAG analyses were performed at each time point of 2 and 6 weeks. Scaffold seeded AF cells were implanted into the back of athymic nude mouse to observe the difference of formation of disc-like tissue depending on pore size in vivo. We confirmed that scaffold with $180{\sim}250{\mu}m$ pores displayed high cell viability in vitro and produced higher ECM than scaffold with other pore sizes in vivo.