• Archives of Plastic Surgery
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• v.34 no.2
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• pp.141-148
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• 2007

Purpose: The object of this study was to evaluate the development of continuous osteogenic differentiation and bone formation after the subcutaneous implantation of the tissue-engineered bone, in vitro. Methods: Human adipose-derived stem cells were obtained by proteolytic digestion of liposuction aspirates. Adipose-derived stem cells were seeded in PLGA scaffolds after being labeled with PKH26 and cultured in osteogenic differentiation media for 1 month. The PLGA scaffolds with osteogenic stimulated adipose-derived stem cells were implanted in subcutaneous layer of four nude mice. Osteogenesis was assessed by RT-PCR for mRNA of osteopontin and bone sialoprotein(BSP), and immunohistochemistry for osteocalcin, and von Kossa staining for calcification of extracellular matrix at 1 and 2 months. Results: Implanted PLGA scaffold with adipose-derived stem cells were well vascularized, and PLGA scaffolds degraded and were substituted by host tissues. The mRNA of osteopontin and BSP was detected by RT-PCR in both osteogenic stimulation group and also osteocalcin was detected by immunohistochemistry at osteogenic stimulation 1 and 2 months, but no calcified extracellular deposit in von Kossa stain was found in all groups. Conclusion: In vivo, it could also maintain the characteristics of osteogenic differentiation that adipose-derived stem cells within PLGA scaffold after stimulation of osteogenic differentiation in vitro, but there were not normal bone formation in subcutaneous area. Another important factor to consider is in vivo, heterologous environment would have negative effect on bone formation as.[p1]

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.735-745
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• 2024

Avian influenza is a serious threat to both public health and the poultry industry worldwide. This respiratory virus can be combated by eliciting robust immune responses at the site of infection through mucosal immunization. Recombinant probiotics, specifically lactic acid bacteria, are safe and effective carriers for mucosal vaccines. In this study, we engineered recombinant fusion protein by fusing the hemagglutinin 1 (HA1) subunit of the A/Aquatic bird/Korea/W81/2005 (H5N2) with the Bacillus subtilis poly γ-glutamic acid synthetase A (pgsA) at the surface of Lactobacillus casei (pgsA-HA1/L. casei). Using subcellular fractionation and flow cytometry we confirmed the surface localization of this fusion protein. Mucosal administration of pgsA-HA1/L. casei in mice resulted in significant levels of HA1-specific serum IgG, mucosal IgA and neutralizing antibodies against the H5N2 virus. Additionally, pgsA-HA1/L. casei-induced systemic and local cell-mediated immune responses specific to HA1, as evidenced by an increased number of IFN-γ and IL-4 secreting cells in the spleens and higher levels of IL-4 in the local lymphocyte supernatants. Finally, mice inoculated with pgsA-HA1/L. casei were protected against a 10LD50 dose of the homologous mouse-adapted H5N2 virus. These results suggest that mucosal immunization with L. casei displaying HA1 on its surface could be a potential strategy for developing a mucosal vaccine against other H5 subtype viruses.

• Macromolecular Research
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• v.14 no.5
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• pp.510-516
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• 2006

Lactide was produced from oligomeric PLA by back-biting reaction of the OH end groups. For optimization of the reaction conditions, the effects of temperature, pressure, PLA molecular weight, and catalyst type on the lactide synthesis were examined. The fraction of D,L-lactide decreased with increasing temperature. Among the various Sn-based catalysts, the D,L-lactide fraction was maximized when SnO was used. A higher yield with lower racemization was observed at lower pressure. The conversion of PLA was maximized at an oligomeric PLA molecular weight of ca. 1380. The yield of lactide increased but the fraction of D,L-lactide decreased with increasing molecular weight. The highest conversion with the lowest racemization degree was obtained at a catalyst concentration of 0.1 wt%. The lactide was more sensitive to racemization because of the entropic effect.

• Journal of Pharmaceutical Investigation
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• v.40 no.4
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• pp.245-250
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• 2010

Porous poly(lactic-co-glycolic acid) microspheres (PLGA MS) have been utilized as an inhalation delivery system and a matrix scaffold system for tissue engineering. Here, gelatin (type A) is introduced as an extractable pH-responsive porogen, which is capable of controlling the porosity and pore size of PLGA microspheres. Porous PLGA microspheres were prepared by a water-in-oil-in-water ($w_1/o/w_2$) double emulsification/solvent evaporation method. The surface morphology of these microspheres was examined by varying pH (2.0~11.0) of water phases, using scanning electron microscopy (SEM). Also, their porosity and pore size were monitored by altering acidification time (1~5 h) using a phosphoric acid solution. Results showed that the pore-forming capability of gelatin was optimized at pH 5.0, and that the surface pore-formation was not significantly observed at pHs of < 4.0 or > 8.0. This was attributable to the balance between gel-formation by electrostatic repulsion and dissolution of gelatin. The appropriate time-selection between PLGA hardening and gelatin-washing out was considered as a second significant factor to control the porosity. Delaying the acidification time to ~5 h after emulsification was clearly effective to make pores in the microspheres. This finding suggests that the porosity and pore size of porous microspheres using gelatin can be significantly controlled depending on water phase pH and gelatin-removal time. The results obtained in this study would provide valuable pharmaceutical information to prepare porous PLGA MS, which is required to control the porosity.

• Journal of the Korean Recycled Construction Resources Institute
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• v.8 no.2
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• pp.204-211
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• 2020

Poly-Lactic Acid(PLA) fiber is an eco-friendly material and is biodegradable, so it can be utilized for manufacturing porous construction materia ls with interna l pore connection. In this study, domestic PLA fiber products(0.5mm india meter, 1.0mm in length, 10mm in length) were tested for melting at high temperatures and high alkality, and they were incorporated with FA-based geopolymer. Compressive strength was obtained through high temperature curing and alkali activator, however the complete melting of the PLA fiber was not ensured. The previous study handling PLA fiber with 0.003mm in diameter was completely dissolved, but 0.5mm and 1.0mm in diameter showed 42.5% and 33.3% of dissolution ratio, respectively. In addition, the increasing fiber volume led floating fibers during curing, which had a negative effect on its workability and solubility. Although the properties of PLA fiber may vary depending on the raw materials and production conditions, PLA fiber with 0.1mm or less diameter is recommended for porous construction material.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.182-191
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• 2007

Bioabsorbable devices have been utilized and experimented in many aspects of orthopaedic surgery. Depending upon their constituent polymers, these materials can be tailored to provide sufficient rigidity to allow bone healing, retain mechanical strength for certain period of time, and then eventually begin to undergo degradation. The objective of this study was to estimate extent in which Poly-L-latic acid (PLLA) implants had bioabsorbability and biocompatibility with bone and soft tissue in dogs and also to develop bioabsorbable, biocompatible materials with the appropriate strength and degradation characteristics to allow for regular clinical use for treating orthopedic problems in humans as well as animals. Eighteen dogs were used as experimental animals and were inserted two types of PLLA implants. PLLA rods were inserted into subcutaneous tissue of back or the abdomen wall. And the rods were tested for material properties including viscosity, molecular weight, melting point, melting temperature, crystallinity, flexural strength, and flexural modulus over time. PLLA screws were inserted through cortical bone into bone marrow in the femur of the dogs and stainless steel screw was inserted in the same femur. Radiographs were taken after surgery to observe locations of screw. Histological variations including cortical bone response, muscular response, bone marrow response were analyzed over the time for 62weeks. The physical properties of PLLA rods had delicate balances between mechanical, thermal and viscoelastic factors. PLLA screws did not induce any harmful effects and clinical complications on bone and soft tissue for degradation period. These results suggest that PLLA implants could be suitable for clinical use.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1638-1642
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• 2012

A series of poly (lactic acid) (PLA) and poly (ethylene glycol) (PEG) tri and multiblock copolymers with relatively high molecular weights were synthesized through the coupling reaction between the bis(acyl chloride) of carboxylated PLA and mono or dihydroxy PEG. The coupling reaction and the copolymer structures were monitored by nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC). The melting temperature (Tm) of PEG blocks decreased with the presence of PLA sequences attaching to PEG blocks. The CMC values were determined to be 10-145 mg/L depending on the length of PLA and PEG blocks and the structure of the block copolymers.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.92-99
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• 2012

The optimal physical factors affecting enzyme production in an airlift fermenter have not been studied so far. Therefore, the physical parameters such as aeration rate, pH, and temperature affecting PLA-degrading enzyme production by Actinomadura keratinilytica strain T16-1 in a 3 l airlift fermenter were investigated. The response surface methodology (RSM) was used to optimize PLA-degrading enzyme production by implementing the central composite design. The optimal conditions for higher production of PLA-degrading enzyme were aeration rate of 0.43 vvm, pH of 6.85, and temperature at $46^{\circ}C$. Under these conditions, the model predicted a PLA-degrading activity of 254 U/ml. Verification of the optimization showed that PLA-degrading enzyme production of 257 U/ml was observed after 3 days cultivation under the optimal conditions in a 3 l airlift fermenter. The production under the optimized condition in the airlift fermenter was higher than un-optimized condition by 1.7 folds and 12 folds with un-optimized medium or condition in shake flasks. This is the first report on the optimization of environmental conditions for improvement of PLA-degrading enzyme production in a 3 l airlift fermenter by using a statistical analysis method. Moreover, the crude PLA-degrading enzyme could be adsorbed to the substrate and degraded PLA powder to produce lactic acid as degradation products. Therefore, this incident indicates that PLA-degrading enzyme produced by Actinomadura keratinilytica NBRC 104111 strain T16-1 has a potential to degrade PLA to lactic acid as a monomer and can be used for the recycle of PLA polymer.

• Polymer(Korea)
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• v.30 no.2
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• pp.168-174
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• 2006

To utilize as highly functional scaffolds for tissue engineering by improving hydrophobicity and cell compatibility of the exist polymer scaffolds, the biodegradable poly(L-lactic acid) (PLLA) films and scaffolds having the optimal hydrophilicity were prepared by in situ plasma treatment and grafting of a carboxyl acid-containing monomer, acrylic acid (AA) in the chamber. From the results of surface analyses, surface-modified nonporous PLLA film and dual pore scaffold surfaces showed high hydrophilicity due to the decrease in contact angle and the increase in carboxylic groups as compared with untreated PLLA control. In particular, among various surface modification methods, Ar(argon)+AA+AA sample prepared by Ar plasma and then acrylic acid treatments displayed lower contact angle and more carboxylic groups thar Ar/AA and Ar+TP(thermal polymerization) samples, indicating that Ar+AA+AA sample was optimally treated for improving its hydrophilicity. In the cases of surface modified nonporous PLLA films and dual pore scaffolds, the adhesion and proliferation of chondrocytes increased with increasing their hydrophilicity.

• Development and Reproduction
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• v.15 no.2
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• pp.87-98
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• 2011

The present experiment was performed to evaluate the osteogenic differentiation of human adipose tissue derived mesenchymal stem cells (ATMSCs) seeded in bioceramic-poly D,L-latic-co-glycolic acid (PLGA) scaffold. Osteogenic differentiation of ATMSCs were induced using the osteogenic induction (OI) medium. ATMSCs were cultured with OI medium during 28 days in well plate. The proliferation of ATMSCs in OI medium group was significantly increased for 14 days of plate culture but slowed after 21 days. On the other hand, proliferation in the control group showed constant increase for 28 days of culturing. The alkaline phosphatase (ALP) activity of ATMSCs in OI medium group increased during the 21 days of culture but decreased on 28 days. However, in control group ALP activity of ATMSCs was continuously decreased as time goes. Nodule was observed at 21 days of culture in OI medium group and confirmed accumulation of calcium in cell by alizarin red staining. ATMSCs were seeded in PLGA scaffold or in Bioceramic-PLGA scaffold, and cultured with OI medium. ALP activity of ATMSCs by osteoblast differentiation in each scaffold increased on 21 days of culture and decreased rapidly on 28 days. ALP activity of ATMSCs was increased highly in Bioceramic-PLGA scaffold compared to PLGA scaffold on 21 days of culturing. SEM-EDS analysis demonstrated that calcium and phosphate content and Ca/P ratio in Bioceramic-PLGA scaffold increased higher than in PLGA scaffold. Biodegradability of scaffold at 56 days after implantation showed that Bioceramic-PLGA scaffold was more biodegradable than PLGA scaffold. The results demonstrated that the differentiation of ATMSCs to osteoblast were more effective in scaffold culture than well plate culture. Bioceramic increased cell adhesion rate on scaffold and ALP activity by osteoblast differentiation. Also, bioceramic was considered to increase the calcium and phosphate in scaffold when ATMSCs was mineralized by osteogenic differentiation. Bioceramic-PLGA scaffold enhanced the osteogenesis of seeded ATMSCs compared to PLGA scaffold.