• IMMUNE NETWORK
• /
• 제11권3호
• /
• pp.163-168
• /
• 2011

Background: Nanoparticles (NPs) prepared from biodegradable polymers, such as poly (D,L-lactic acid-co-glycolic acid) (PLGA), have been studied as vehicles for the delivery of antigens to phagocytes. This paper describes the preparation of antigen-loaded PLGA-NPs for efficient cross-priming. Methods: NPs containing a similar amount of ovalbumin (OVA) but different sizes were produced using a micromixer-based W/O/W solvent evaporation procedure, and the efficiency of the NPs to induce the cross-presentation of OVA peptides were examined in dendritic cells (DCs). Cellular uptake and biodistribution studies were performed using fluorescein isothiocyanate (FITC)-loaded NPs in mice. Results: The NPs in the range of $1.1{\sim}1.4{\mu}m$ in size were the most and almost equally efficient in inducing the cross-presentation of OVA peptides via $H-2K^b$ molecules. Cellular uptake and biodistribution studies showed that opsonization of the NPs with mouse IgG greatly increased the percentage of FITC-positive cells in the spleen and lymph nodes. The major cell type of FITC-positive cells in the spleen was macrophages, whereas that of lymph nodes was DCs. Conclusion: These results show that IgG-opsonized PLGA-NPs with a mean size of $1.1{\mu}m$ would be the choice of biodegradable carriers for the targeted-delivery of protein antigens for cross-priming in vivo.

• Macromolecular Research
• /
• 제13권5호
• /
• pp.446-452
• /
• 2005

The interactions between the surface of scaffolds and specific cells play an important role in tissue engineering applications. Some cell adhesive ligand peptides including Arg-Gly-Asp (RGD) have been grafted into polymeric scaffolds to improve specific cell attachment. In order to make cell adhesive scaffolds for tissue regeneration, biodegradable nonporous poly(L-lactic acid) (PLLA) films were prepared by using a solvent casting technique with chloroform. The hydrophobic PLLA films were surface-modified by Argon plasma treatment and in situ direct acrylic acid (AA) grafting to get hydrophilic PLLA-g-PAA. The obtained carboxylic groups of PLLA-g-PAA were coupled with the amine groups of Gly-Arg-Asp-Gly (GRDG, control) and GRGD as a ligand peptide to get PLLA-g-GRDG and PLLA-g-GRGD, respectively. The surface properties of the modified PLLA films were examined by various surface analyses. The surface structures of the PLLA films were confirmed by ATR-FTIR and ESCA, whereas the immobilized amounts of the ligand peptides were 138-145 pmol/$cm^2$. The PLLA surfaces were more hydrophilic after AA and/or RGD grafting but their surface morphologies showed still relatively smoothness. Fibroblast adhesion to the PLLA surfaces was improved in the order of PLLA control

• Journal of Periodontal and Implant Science
• /
• 제29권3호
• /
• pp.507-519
• /
• 1999

The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

• Macromolecular Research
• /
• 제16권8호
• /
• pp.695-703
• /
• 2008

Nanoparticles have tremendous potential in cancer prevention, detection and augmenting existing treatments. They can target tumors, carry imaging capability to document the presence of tumors, sense pathophysiological defects in tumor cells, deliver therapeutic genes or drugs based on the tumor characteristics, respond to external triggers to release an appropriate agent, document the tumor response, and identify the residual tumor cells. Nanoparticles < 30 nanometers in diameter show unexpected and unique properties. Furthermore, particles < 5 nanometers in size can easily penetrate cells as well as living tissues and organs. This study evaluated the safety of nano materials in a living body and the relationship between the living tissue and synthetic nano materials by examining the in-vitro cytotoxicity of poly(lactic-co-glycolic) acid (PLGA) nano-spheres and fluorescein isothiocynate(FITC)-labeled dendrimers as polymer nanoparticles. PLGA was chosen because it has been used extensively for biodegradable nanoparticles on account of its outstanding bio-compatibility and its acceptance as an FDA approved material. The dendrimer was chosen because it can carry a molecule that recognizes cancer cells, a therapeutic agent that can kill those cells, and a molecule that recognizes the signals of cell death. Cytotoxicity in L929 mouse fibroblasts was monitored using MTT assay. Microscopic observations were also carried out to observe cell growth. All assays yielded meaningful results and the PLGA nanoparticles showed less cytotoxicity than the dendrimer. These nano-particles ranged in size from 10 to 100 nm according to microscopy and spectroscopic methods.

• IMMUNE NETWORK
• /
• 제21권6호
• /
• pp.44.1-44.15
• /
• 2021

Tumor peptides associated with MHC class I molecules or their synthetic variants have attracted great attention for their potential use as vaccines to induce tumor-specific CTLs. However, the outcome of clinical trials of peptide-based tumor vaccines has been disappointing. There are various reasons for this lack of success, such as difficulties in delivering the peptides specifically to professional Ag-presenting cells, short peptide half-life in vivo, and limited peptide immunogenicity. We report here a novel peptide vaccination strategy that efficiently induces peptide-specific CTLs. Nanoparticles (NPs) were fabricated from a biodegradable polymer, poly(D,L-lactic-co-glycolic acid), attached to H-2K^b molecules, and then the natural peptide epitopes associated with the H-2K^b molecules were exchanged with a model tumor peptide, SIINFEKL (OVA_257-268). These NPs were efficiently phagocytosed by immature dendritic cells (DCs), inducing DC maturation and activation. In addition, the DCs that phagocytosed SIINFEKL-pulsed NPs potently activated SIINFEKL-H2K^b complex-specific CD8⁺ T cells via cross-presentation of SIINFEKL. In vivo studies showed that intravenous administration of SIINFEKL-pulsed NPs effectively generated SIINFEKL-specific CD8⁺ T cells in both normal and tumor-bearing mice. Furthermore, intravenous administration of SIINFEKL-pulsed NPs into EG7.OVA tumor-bearing mice almost completely inhibited the tumor growth. These results demonstrate that vaccination with polymeric NPs coated with tumor peptide-MHC-I complexes is a novel strategy for efficient induction of tumor-specific CTLs.

• 폴리머
• /
• 제24권4호
• /
• pp.505-512
• /
• 2000

본 연구는 초임계 용매인 $CO_2$, CHF$_3$ 및 CHClF$_2$내에서 poly(lactide-co-glycolide) [PLGAI 용액과의 흔합물 밀도를 측정하였다. 초임계 용매와 Poly(lactic acid) [PLA] 및 PLGA간의 흔합물 밀도는 온도 27-10$0^{\circ}C$와 압력 3000 bar까지 실험하여 나타내었다 [PLGA$_{x}$의 X는 0~50 mo1% 범위에 대한 glycolide의 몰농도이다]. PLA-$CO_2$ 흔합물은 약 1430 bar 이내에서, PLA-CHF$_3$계는 700 bar 이하에서, PLA-CHClF$_2$계는 100bar 이하에서 각각 용해되었다. 이때 온도범위는 27~93$^{\circ}C$이며, 흔합물 밀도는 1.084~l.334g/$cm^3$ 범위에서 나타났다. PLGA$_{15}$ 공중합체-$CO_2$ 흔합물은 약 1900 bar 이하에서 용해되었으며, 이때 혼합물 밀도는 37~92$^{\circ}C$에서 1.158~l.247g/$cm^3$으로 나타났다. PLGA$_{25}$공중합체-$CO_2$계는 약 2390 bar이하에서, PLGA$_{25}$-CHF$_3$계에 대해서는 1470 bar이하에서, PLGA$_{25}$-CHClF$_2$계에 대해서는 118 bar 이하에서 각각 용해되었으며, 흔합물 밀도는 29~81$^{\circ}C$사이에서 1.154~1.535g/$cm^3$로 나타났다. PLGA$_{50}$-$CO_2$계는 24$0^{\circ}C$, 3000 bar내fl서는 용해되지 않았으며, 반면 PLGA$_{50}$과 CHClF$_2$의 흔합물은 오히려 5$0^{\circ}C$와 100 bar내에서 쉽게 용해되었다. 또한 PLGA와 CHClF$_2$계는 glycolide 농도가 증가함에 따라 흔합물 밀도가 증가하였다.다..다..다..

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권20호
• /
• pp.8693-8698
• /
• 2014

Background: Up-regulation of hsp90 gene expression occurs in numerous cancers such as lung cancer. D,L-lactic-co-glycolic acid-poly ethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG may inhibit the expression. The purpose of this study was to examine whether nanocapsulating 17DMAG improves the anti cancer effect over free 17DMAG in the A549 lung cancer cell line. Materials and Methods: Cells were grown in RPMI 1640 supplemented with 10% FBS. Capsulation of 17DMAG is conducted through double emulsion, then the amount of loaded drug was calculated. Other properties of this copolymer were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity on the grown of lung cancer cell line was carried out through MTT assay. After treatment, RNA was extracted and cDNA was synthesized. In order to assess the amount of hsp90 gene expression, real-time PCR was performed. Results: In regard to the amount of the drug load, IC50 was significant decreased in nanocapsulated(NC) 17DMAG in comparison with free 17DMAG. This was confirmed through decrease of HSP90 gene expression by real-time PCR. Conclusions: The results demonstrated that PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of hsp90 expression by enhancing uptake by cells. Therefore, PLGA-PEG could be a superior carrier for this kind of hydrophobic agent.

• Journal of Periodontal and Implant Science
• /
• 제35권3호
• /
• pp.549-561
• /
• 2005

이 연구의 목적은 새로이 제작된 chitosan-poly(L-lactic acid)(PLLA) 다층 다공성 차폐막의 생체적합성 및 골세포활성도 및 골재생능을 평가하는 것이다. 제작된 차폐막을 24 well에 넣고 clonal osteoblast-like cell line(MC3T3-E1)을 접종한 군을 실험군으로, 차폐막을 사용하지 않은 대조군으로 하였다. 배양 1일, 7일 및 14일째에 각 well에서 세포수를 측정하였다. 주사전자현미경을 이용하여 차폐막에 부착된 세포의 형태관찰을 시행하였다. RNA 추출 및 RT-PCR을 실시한 후, agarose gel상에서 전기영동하여 조골세포 표식자인 collagen type I(COL), osteopontin(OP) 및 osteocalcin(OC) mRNA의 발현을 관찰하였다. 제작된 매트릭스의 생체적합성 및 골재생능을 관찰하기 위하여 백서의 두개골에 직경 8mm의 원형 결손부를 형성한 후 차폐막을 이식한 군을 실험군으로, 아무 것도 넣지 않은 군을 대조군으로 하여 4주 경과 후 실험동물을 희생시킨 후 조직학적관찰을 시행하였다. 시간경과에 따른 부착세포수 관찰결과, 배양 14일까지 조골세포의 수가 지속적으로 증가하였고, 주사전자현미경으로 세포의 형태 관찰결과, 배양된 세포들은 중층의 형태로 성장하면서 시간경과에 따라 세포가 응집되는 양상을 나타내었다. 관찰 기간동안 COL, OP, 및 OC mRNA의 발현이 관찰되어 배양 전 기간동안 조골세포의 형질이 잘 유지되고 있음을 알 수 있었다. 백서 두개골 결손부에 이식된 차폐막은 염증반응 없이 주위 조직과 우수한 생체적합성을 나타내었으며, 차폐막을 이식하지 하지 않은 대조군에 비해 높은 신생골 형성을 나타내었다. 이상의 관찰결과로 새로이 제작된 chitosan-PLLA 차폐막은 우수한 생체적합성 및 골재생능을 나타냄을 알 수 있었으며, 향후 이를 골조직 재생 및 치주조직유도재생 분야에 응용될 수 있을 것으로 생각된다.