Earticle

The use of L-carbohydrates and their corresponding nucleosides in medicinal application has greatly increased. For example L-ribose has been much in demand as the starting material for curing hepatitis B. However, L-ribose dose not occur in nature. L-ribose was reported to be produced by epimerization. Bilik published a simple synthesis of L-ribose by molybdate-catalyzed epimerization of the readily available L-arabinose in 1973. The reaction now is known as the Bilik reaction. In this study, the effects of the temperature, solvent, reaction time and molybdenum(Ⅵ) oxide amount on epimerization were examined by measuring the product concentration in sugar HPLC column. In addition, conversion mixture of L-ribose, L-arabinose, and MoO3 was separated by using ion exchange cloumn packed by Dowex ca resin.

Melissa officinalis is perennial herb occurring throughout the east Asia, which have been used for various traditional medicinal treatment. This plant is known to mainly possess rosmarinic acid and protocatechuic acid as polyphnolic acids. To investigate the biological activity of the flavonoids in these plants, it is necessary to develop an effective system for extracting and purifying the flavonoids with possible biological functions. Therefore, in this study, the extraction process with operating variable such as temperature, solvent type and concentration, and solid to solvent ratio was optimized by response surface analysis methodology. Also, the chromatographic purification of rosmarinic acid in the extract obtained from the solvent extraction was carried out by using adsorption chromatography resins such as XADs and Sephadex LH-20 at a variety of conditions. In conclusion, this work is used as basic information for developing an effective purification strategy of rosmarinic acid from plant resources.

Herein, we propose a novel methodology for efficient recovery of cell-free synthesized recombinant proteins. Through the expression of His-tagged proteins in the presence of Ni-NTA resin, the nascent polypeptides were instantly immobilized on the solid surface, thereby avoiding the inter-molecular aggregation. Upon the elution with imidazole, compared to the reactions conducted without the resin, it was found that substantially higher amounts of soluble proteins were recovered most likely through improved folding the nascent polypeptides on the surface of resin. We expect that the proposed method will significantly contribute to rapid generation of properly folded proteins for numerous applications.

Sun protecting substances are capable of protecting humans from harmful effects solar radiation such as aging and skin cancer (1, 2). Bisethylhexyloxyphenolmethoxyphenyltrizine(BEMT) is one of the most widely used oil-soluble UVA+UVB absorber in sunscreen cosmetic product. Stratum corneum is the major barrier in percutaneous absorption process (3, 4). The purpose of this study is to confirm the skin delivery property and synergistic UV-blocking effect of solid lipid nano-particles containing the BEMT with a sun screen agent. The study of skin permeability is important, and performed by various methods. In this study, target active ingredient is BEMT. I compared in vitro release and penetration of this active ingredient in O/W emulsion and SLN formulation. According to Wissing, The release rate of oxybenzone is decreased when using SLN formulations compared emulsions (5). As a result, in vitro penetration and release of BEMT is generally higher O/W emulsion than the SLN formulation. After having a 24 hours, cumulative transdermal absorption amount of BEMT is higher emulsion(16.04㎍/㎠) than SLN formulation(4.21㎍/ ㎠). The other hand, in vivo study, penetration of BEMT into the skin was investigated by the stripping method. It was shown that the rate of release could be decreased by 80% in the SLN formulation. SLN matrix sustained release carrier system, therefore sunscreen agent remain longer on the skin. And the occlusive effect of SLN was physical sunscreen with increased synergistic UV-blocking effect (6). In vitro UV protection result, the sun protection factor (SPF) could be increased by 100% in the SLN formulation. Therefore, BEMT-SLN led synergistic UV-blocking behavior. The BEMT residual content gradually decreased to 80.9% over 12 hours the result of UV radiation, but the residual content of BEMT with an encapsulated solid lipid decreased to only 92.3%. Also the BEMT-SLN led to improve of photochemical stability effect on ultra violet. Ultimately it appeared that Solid lipid nanoparticle was the most effective to adopt BEMT and may be used for a useful drug delivery system in the sun care product.

In ion exchange chromatography for the recovery of potassium clavulanate from fermentation broth, a large amount of potassium chloride existed in the resulting solutions. In this study, diafiltration with nanofiltration membrane was employed to retain potassium clavulanate while removing chloride ions from the solutions. In the diafiltration of a solution containing 2.5 mM of potassium clavulanate and 1,000 mM of potassium chloride, almost complete removal of chloride was possible with no significant loss of clavulanate. However, clavulanate degradation was observed, although not very serious. Based on the experimental results of diafiltration, three different mathematical models were developed. The effect of temperature on clavulanate degradation was also modeled and was incorporated into those diafiltration models. Model predictions of clavulanate concentration and chloride concentration were in good agreement with the experimental data.

To make more functional scoria powder into cosmetic material, studies on the supercritical impregnation of α-tocopherol into Jeju scoria were made. Supercritical carbon dioxide is non-toxic and more penetrative than organic solvent and easy to remove. Experiment was composed of three steps, sterilization of scoria powder, supercritical impregnation of α-tocopherol into scoria and HPLC analysis of impregnated scoria. The supercritical impregnation was performed at pressure 400bar and temperature 60℃. The amount of equilibrium impregnation increased with solubility of α-tocopherol in supercritical carbon dioxide. Analysis of impregnated α-tocopherol was HPLC. analysis condition was acetonitrile:methanol (75:25) 1.5ml/min at UV 220nm.

Protein은 Acetic Acid가 0%일 때를 제외하고 그 이상의 농도에서는 거의 영향을 받지 않았고, Collagen은 Acetic Acid의 증가에 따라 난막으로부터 추출되는 양이 많아졌다. 그러나 Protein과 Collagen모두 Cysteine의 증가에 따라 추출양이 많아지다가 적정농도 이상에서는 다시 추출양이 감소하는 경향을 볼 수 있었다. 이는 Cysteine이 과량으로 첨가될 경우 sulfide결합을 끊는데 쓰이고 남은 Cysteine이 한정된 샘플 안에 존재하여 상대적으로 Collagen의 양이 적기 때문인 것으로 생각된다. 또 낮은 온도나 짧은 시간 조건에서는 반응성 저하로 이들 요인이 거의 영향을 미치지 않았다.

The objective of this study is to investigate the optimum conditions of extracting collagens from eggshell membranes. The effect of time, temperature, and the concentration of cysteine and acetic acid content on the collagen extraction has been studied. UV-Visible spectrophotometer was used to quantify the protein and collagen extraction. Collagen extraction is considerably affected by both of the concentration acetic acid and cysteine. However, protein extraction is affected by the concentration of cysteine rather than that of acetic acid.

Pipefish, Syngnathus schlegeli (PF) a marine teleost fish, is well known not only for its special medicinal composition and used as one of the most famous and expensive materials of traditional Chinese medicine. It was extracted with water (PFW), methanol (PFM) and ethanol (PFE), respectively and evaluated by various antioxidant assays. The including total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, and alkyl radical scavenging. Further, the ROS level was detected using a fluorescence probe, 2’,7’-dichlorofluorescin diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on human fibrosarcoma cell (RAW264.7). Those various antioxidant activities were compared tostandard eantioxidants such as α -tocopherol. Among PFM exhibited the highest antioxidant activity in linoleic acid system, effective reducing power, DPPH radical scavenging, hydroxyl radical scavenging, superoxide radical scavenging, alkyl radical scavenging, inhibitory intracellular ROS. Furthermore, MTT assay showed no cytotoxicity on human embryonic lung fibroblasts cell line (MRC-5) and Murine Leukemia Macrophage cell line (RAW264.7). This antioxidant property depends on concentration and increasing with increased amount of sample.

In this study, biochemical characterization including hemagglutination of erythrocytes, molecular weight, optimum temperature, thermal stability, optimum pH, carbohydrate specificity, and inhibitory effect of metal ion were studied in lectin of eggplant (Solanum melongena L.) fruit prepared by ammonium sulfate fractionation and affinity chromatography. This lectin was agglutinated by trypsin-treated rat blood erythrocyte. The molecular weight of this lectin by SDS-PAGE was estimated to be approximately 19.3 kDa of a single band. This lectin have no activity by cabohydrates such as D-glucose etc. The optimum range of temperature and pH were 10-20℃ and pH 6.2-7.2, respectively. This lectin was relatively stable at 20-70℃. And the activity of this lectin was not inhibited by Ca2+, Co2+, Cu2+, Fe2+, Mg2+, and Mn2+.

Bioactive sterol metabolites, fucosterol (1) and ergosterol (2), were obtained for the first time from the marine brown alga Ecklonia cava. Their structures were clearly elucidated by the comprehensive analyses of the NMR spectral data. Compound (1) showed the various potential bioactivities, such as anti-fungal, anti-bacteria and anti-oxidant activities.

Because of the visible nature of dermatologic diseases such as melasma, lentigines, they have a considerable psychological effect on affected patients. To treat these diseases, a number of skin whitening compounds have been screened for their effectiveness in reducing melanogenesis but none are satisfactory. To developing new and safe skin whitening agents from natural sources, Chinese plants have been evaluated. From screening results, Lasa B29 inhibited 43.7% melanin synthesis of murine B16F10 melanoma cells and exhibited low cytotoxicity (8.1%) at a concentration of 100 ug/ml. To find the mechanism of melanin inhibition, inhibitory activity on tyrosinase, the key enzyme of melanogenesis, was further evaluated. The results showed inhibitory effects on the activity of intracellular tyrosinase (34.7% at 100 ug/ml) but not the mushroom tyrosinase. To isolate an effective compound, silica open column chromatography, and preparative high performance liquid chromatography were used.

Flavors are complex mixtures of comparatively volatile substances and labile components of which the sensory perception can be changed as a result oxidation, chemical interactions or volatilization. Cinnamomonn zeylanicum essential oil is a widely used spicy and have many applications in perfumery, flavoring and pharmaceutical industries. Antioxidant activity of reconstituted liposomal incorporation of Cinnamomonn zeylanicum essential oil was studied for application to cosmetic industry. Liposomal formulations were characterized by transmission electron microscopy(TEM) and optical and light polarized microscopy for vesicle formation and morphology, and by dynamic laser light scattering for size distribution. Free radical-scavenging capacities were also evaluated with the test of 2,2-diphenyl-1-picryhydrazylradical (DPPH*), spectrophotometrically at 517nm. Results showed that Cinnamomonn zeylanicum essential oil can be incorporated in good amounts in the prepared vesicular dispersions. Liposomal incorporation of Cinnamomonn zeylanicum essential oil has spherical structure and 20~80nm size. Cinnamomonn zeylanicum (EDA; 92.15% sc50; 2.5 ㎕/ml) showed good antioxidative activity. we expect the potential activities of the Liposomal incorporation of Cinnamomonn zeylanicum essential oil as a material of functional cosmetics.

Soluble N-ethylmaleimide sensitive factor attachment receptor (SNARE) proteins play crucial role in membrane fusion and exocytosis. SNARE proteins are believed to generate fundamental force that is needed to facilitate the merging of two opposing membranes by forming so called “SNARE complex.” Inhibition of neuronal SNARE complex formation impairs neurotransmitter release. In the present study, we carried out to examine the effects of synthetic peptides patterned after the α–helical region of the SNARE motifs on SNARE complex formation and neuronal exocytosis. Thirteen 17-mer synthetic peptides were assayed for their abilities to prevent SNARE-driven membrane fusion. Among them, a few synthetic peptides inhibited the membrane fusion by ~70%. Notably, peptides patterned after the N-terminal region of the SNARE motifs had a stronger effect on membrane fusion than those of middle and C-terminal domains. The found peptide might function as anti-wrinkle agent by blocking SNARE complex formation while Botox cleaves SNARE proteins. Further, the found peptide might be developed to potential pain-killer.

There is a growing interest in finding skin depigmenting agents from natural sources in the cosmeceutical and pharmaceutical markets. In this study, methanol extracts of 57 traditional medicinal plants growing in Vietnam and 45 plants growing in China were evaluated. Cell-based and non-cell-based assays were performed to measure (i) melanin contents, (ii) cell viability, (iii) antioxidant activity, and (iv) tyrosinase inhibitory activity. The screening results revealed that 14 Vietnamese plants and eight Chinese plants had high potentials as skin whitening agents. For further isolated, S1 sample was chosen. Its methanol extract exhibited a 32.91% and 64.93% inhibition of melanin synthesis on B16F10 cells and Melan-a cell without cytotoxicity at a concentration of 200µg/ml. Moreover, this extract inhibited mushroom tyrosinase as well as intracellular tyrosinase and has high antioxidant activity. To elucidate the mechanism of S1–induced depigmenting activity, the protein level of tyrosinase was also determined by Western blotting. These results suggested that S1 sample could be a high potential candidate as a skin depigmenting agent in the cosmeceutic and pharmaceutical industries.

Angelica Gigas extracts were prepared by water boiling, supercritical fluid, and ethanol extraction. Supercritical fluid extract was performed at pressure of 200-400 bar and at temperature of 40-70℃. Inhibitory effect and mechanism of skin aging by Angelica Gigas extracts were investigated using proteome analysis. The human fibroblast were cultured in Dulbecco's Modified Eagle's Medium(DMEM) with or without extracts. The proteome of cultured human fibroblast were profiled using 2D-PAGE. Isoelectric focusing was carried out using commercial immobiline dry strips 3-10L 13cm. SDS-PAGE was performed using 18x24cm, 11% acrylamide gel1). Gels were stained with silver nitrate, analyzed by image master 2D software program. In conclusion, we could identify that many factors such as collagen, integrin beta-1, which affect the skin aging, were regulated by Angelica Gigas extracts.

Nails are continuously damaged and injured during our lifetime. Recently artificial nail art procedures can give severe damage to the natural nail. Thus, nail guard (also called nail strengthener/hardener) should be used to protect the nail from any artificial nail treatments. Experiments were intended to study the effect of nail guards products on the surface and cut section of the damaged nail.1) Three cosmetical nail guards products were selected considering the price, ease of operation and popularity. The group members applied the nail guard everyday to the damaged nail for two weeks. The morphological changes of nail surface and a cutting plane were monitored and analyzed using scanning electron microscope (SEM). Many side effects of nail art treatment have been confirmed by the morphological change and questionnaire survey. The damaged portion of the upper part is more severe than that of the lower part. The lamella structure of the damaged nail showed the serious crack. The nail guards products gave a temporal strengthening effect to the damaged nail. Among the ingredients of the nail guards products, the protein and moisturizing components are considered to be participated in the hardening of the nail.

Camellia oil, citron oil, and seaweed extract were used to develop functionally active natural materials for permanent wave. Scanning electron microscopy (SEM), tensile strength, and the rate of hair expansion were monitored to determine the permanent efficiency and hair damage. Hair was damaged by permanent wave treatment with thioglycolate and L-cystein. Hair damage with thioglycolate was more severe than that with L-cystein and it could be minimized after treatment with natural materials. With the increase of heat treatment, the hair damage was also increased. After treatment using camellia oil, the hair damage could be minimized with heat permanent.

Hyaluronic acid (HA) is a linear polysaccharide formed from disaccharide units containing Nacetylglucosamine and glucuronic acid. It is generally known that digestion of hyaluronic acid with the enzyme hyaluronidase introduce a double bond is formed in terminal group of the hydrolyzed HA.1) In this study hyaluronic acid was hydrolyzed into very low molecular weight hyaluronic acid (oligo HA) by ion exchange resin whithout forming a double bond or occurring ring opening. In order to investigate the potential of oligo HA as a cosmetic ingredient, we measured its cytotoxicity in fibroblast and SIRC cell line, moisturizing effect, and the penetration effect using artificial skin and Caco-2 cell line.2) Oligo HA did not showed any cytotoxicity at a concentration of 100 Ag/ml in fibroblast cell but it showed weak proliferation. In vitro ocular test of oligo HA, it showed negligible cytotoxicity up to the 2000 Ag/ml concentration in SIRC cell. In the test of the single and repeated cutaneous applications, oligo HA under occlusive patch did not provoke any cumulative irritation and sensitization.3) Oligo HA at a concentration of 0.01% exhibited a better moisturizing effect than hyaluronic acid at a concentration of 0.01%. In the permeability test using artificial skin and Caco-2 cell line, hyaluronic acid (M.W. 1,100,000) was hardly observed in the down medium of the inserts. On the other hand, Oligo HA (M.W. 5,000) was detected in the down medium up to 14% at 6 hr using Caco-2 cell and the down medium up to 90% at 6hr using artificial skin.4) This result suggests that Oligo HA could be useful as an active ingredient for cosmetics.

In the cascade of neurotransmission, neuronal SNARE proteins play a central role. Neuronal SNARE proteins mediate membrane fusion between presynaptic membrane and synaptic vesicle. By the result of synaptic membrane fusion, a fusion pore is made through which neurotransmitters are released. SNARE complex is thought to generate fundamental force that is required for the membrane fusion. As is well-known, a few types of botulinum neurotoxin have been developed and commercialized for the purpose of cosmetic and therapeutic uses. For example, Botox, a popular cosmetic agent, is to cleave SNARE proteins disabling neurotransmission. So, it is that SNARE folding modulator may act like Botox by inhibiting the folding of SNARE complex not by cleaving the SNARE proteins. In the present study, we could draw some candidate SNARE folding modulators from phenolic compounds. A few compounds inhibited SNARE complex formation. As expected, the phenolic compounds inhibited membrane fusion and neurotransmitter release from PC12 cells, too. Probably, the phenolic compounds may play a role as an effective anti-wrinkle agent.

Phellinus linteus(PL) has been used as a traditional herb medicine for several hundreds years in Asia. Most cancer patients in Korea have used Sang Hwang mushroom as medication.1,2) As a result of the used polymethylmethacrylate (PMMA) with PL, and produced microcapsule, it was able to confirm PL microcapsule through optical microscope, SEM and TEM. The emulsion containing PL microcapsule was stable at temperature conditions (0℃, 25℃, 40℃), artificial sun lamp test and cycle chamber, pH, viscosity during 28 days. Dynamic rheological property was not change on emulsion containing PL microcapsule. To test for the releasing of PL in the skin lotion during 14 days. The present study suggested that PL microcapsule have a potential as a new slow release cosmetic material. From the result of human patch test to assess the safety showed no stimulus of negative reaction on skin. Based on results, we determine that leaves of PL microcapsule can be used as a pharmaceutical and cosmeceutical material in the future.

천연산물들을 UV 영역에서 scanning하여 280-320 nm, 320-400 nmdud역에서 흡수대가 높은 천연산물을 스크리닝했을 때 녹차와 HWG, DAD, BRAS, DOL, NRM, JCW, KEW등이 자외선 영역에서 차단효과가 큰 것으로 나타났으AU, NIH/3T3 세포를 사용하여 UV를 조사후 회복율을 보면, 녹차, WGS, WG, HWG, DCHC 및 AGA 등이 좋은 효과를 나타내어 스킨케어의 중요한 재료로 사용가능함을 보였다. 그리보 보습효과는 DAD, HWG, SUK, AGA, JCW 및 CHIL등의 성분이 보습을 유지하는 효과가 높은 것으로 나타났으며, 아토피를 억제하는 천연산물로서는 BAKJLE, GSE, HWG, DAD, AGA, CHIL, SUK등이 좋은 효과를 보였다. Nude mouse를 사용하여 UV 처리후 피부가 회복되는데는 사용한 천연산물 모두 피부가 원상대로 회복되는 것을 볼 수 있었다. DAD, HWG, AGA, GSE및 DCHC에서 좋은 효과를 보였다.

When UV blocking effect was scanned with spectrophotometer from 320-400nm, greentea, HWG, DAD, BRAS, DOL, NRM, JCW, KEW are effective for protecting UV blocking, and recovery effect after UV irradiation to NIH/3T3 cell, greentea, WGS, WG, HWG, DCHC and AGA were showed higher recovering effect for skin care materials. And animal test of UV irradiation, nude mouse was completely recovered originally by DAD, HWG, AGA, GSE and DCHC. Moisturing effect of natural product with in vitro was showed higher function by DAD, HWG, SUK, AGA, JCW and CHIL. For anti-atophic effect against compound 48/80 is showed a good effect by BAKJLE, GSE, HWG, DAD, AGA, CHIL, and SUK.

To investigate the effect of water extract of Saxifraga stolonifera MEERBURGH on skin care, we measured anti-oxidant and anti-aging activity. Saxifraga stolonifera MEERBURGH water extract itself had anti-oxidant activity in a dose-dependent manner in DPPH radical scavenging. Silica dose-dependently increased the intracellular ROS generation in RAW 264.7 cells. Saxifraga stolonifera MEERBURGH water extract inhibited silica-induced intracellular superoxide anion generation and H2O2 and hydroperoxide generation in RAW 264.7 cells. Saxifraga stolonifera MEERBURGH water extract significantly inhibited both hyaluronidase and elastase activity, also significantly inhibited MMP-1(collagenase) activity as well. In NIH 3T3 fibroblast cells, Saxifraga stolonifera MEERBURGH water extract significantly increased collagen-like polymer synthesis, which suggest that the Saxifraga stolonifera MEERBURGH water extract might be used as hydration and anti-wrinkle agents. From the above results, it is referred that Saxifraga stolonifera MEERBURGH water extract appears to have potent anti-oxidant and anti-aging activity. It is suggested that the main ingredients of stolonifera MEERBURGH water extract play an important role in anti-oxidant and anti-aging activity.

오일 함량에 따른 microemulsion과 nanoemulsion의 size를 비교한 결과 oil의 함량이 높을수록 microemulsion은 입도가 증가한 반면 nanoemulsion에서는 입도가 감소하였다. 오일함량 약25%에서 nanoemulsion의 입도는 약 60nm이었으며, 에멀젼 입자에 의한 광산란이 현격히 감소하여, 마이크로에멀젼과 달리 매우 높은 투명도를 보였다. 또한 nanoemulsion은 4℃-25℃ 범위에서 90일 동안 입도가 거의 변하지 않아 높은 장기 안정성을 나타내었다.

We have investigated the droplet size of nanoemulsion as a function of oil content in o/w emulsion system. On the contrary to microemulsion, the droplet size of nanoemulsion decreased as the content of oil increased. At the oil content of 25 wt% the droplet size of nanoemulsion was about 60nm, highly transparent and very stable until 90 days at the temperature range 4℃ - 45℃.

Defects of articular cartilage can be resolved using mesenchymal stem cells (MSCs) isolated from umbilical cord blood (UCB), which have differentiation capability into chondrocytes.1) In this study, chondrogenesis of MSCs in bioabsorbable membrane was investigated to apply in repairing damaged articular cartilage. MSCs isolated from UCB and characterized by fluorescence-activated cell sorter (FACS) were cultured in bioabsorbable membrane so that they could be expanded in three dimensional environment. Chondrogenesis of MSCs was performed in chondrogenic medium with transforming growth factor-β3 (TGF-β3). The location of MSCs in membrane could be observed by scanning electron microscope (SEM). Moreover, the MSCs in the membrane could also be visualized by their intense green fluorescence using confocal microscopy since enhanced green fluorescent protein (EGFP) was transformed into the MSCs. Chondrogenesis of MSCs was verified by Alcian blue, Safranin O staining and RT-PCR. These results show that MSCs can successfully differentiate into chondrocytes in bioabsorbable membrane. In addition, chondrogenic potential of MSCs can be utilized for the repair of injured articular cartilage by cell-based therapy.

Melanogenesis is a physiological process resulting in the synthesis of melanin pigments, which play a crucial protective role against skin photocarcinogenesis. EtOH extract and various fractions from M. micromalus were analysed using 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO) radicals, superoxide radical, Xanthine oxidase, respectively. And it’s results, EtAOc fraction inhibited the DPPH (IC50 : 1.2g/ml), NO radicals (IC50 : 584.9g/ml), superoxide radical (IC50 : 0.6g/ml), Xanthine oxidase (IC50 : 4.9g/ml) on low concentration. The study was designed to investigate the inhibitory effect of extract from M. micromalus on melanogenesis in Melan-a cells. We also estimated total melanin content as a final product and activity of tyrosinase, a key enzyme, on melanogenesis in Melan-a cells. The melanin content and tyrosinase activity were decreased in extract-treated cells in a dose dependent manner compared to control group. EtAOc fractions of M. micromalus significantly inhibited cell tyrosinase activity and melanin content and this inhibitory action was not due to the cytotoxicity. Taken together, this study suggests that EtAOc fractions of M. micromalus could contribute to the melanogenesis inhibition.

To accomplish the large enhancement in cell number required for the repair of articular cartilage defects, serum is normally supplemented into the expansion medium. However, use of serum has disadvantages such as the risk of disease, lot to lot variations, and the fact that it is difficult to determine what types of substances make up serum makes it unpreferable for use in culture. A chemically-defined medium will alleviate such dilemmas. Maintenance of the redifferentiation potential of the cells is also important to take into account since chondrocytes tend to lose their phenotype when expanded, thereby producing less glycosaminoglycans and changing their collagen synthesis.1) Following a literature search on serum-free media for chondrocytes, information on growth factors and other components required for supplementation was organized. Using these various supplements, a basal serum-free medium is under development which will later undergo screening and optimization via statistical methods such as the Plackett Burman approach. Effects on growth and chondrogenic potential will be observed for chondrocytes cultured in the serum-free medium, commercial chondrocyte medium and serum-containing medium.

Bio capsule molecule, Liposome was prepared by film dispersion method. Skin whitening agent, ‘Arbutin’ was dissolved in water before liposome formation and capsulated by liposome. Sonication and extrusion method were applied for the homogenization of the size and shape of liposome. Changes of liposome size and entrapment efficiency were investigated to confirm liposome stability.1) Arbutin encapsulation was also induced by size extrusion. Extrusion of liposome suspension was serially executed 400, 200, 100nm pore size membrane filter. Arbutin capsulated liposome was separated by superdex-100 column chromatography.2) Then, HPLC analysis was performed to report Entrapment efficiency of arbutin. Entrapment efficiency of pre-dissolved and extursion method were compared.

In order to evaluate the usefulness of its extract as a functional biomaterial, we investigated several biological activities using the ethanol extract and its fractions from Pimpinella komarovii leaves. The inhibitor effect of ethanol extract on tyrosinase activity was higher than water fraction. When 50㎍/㎖of ethyl acetate (EtOAc) fraction was applied, the inhibition ratio of tyrosinase activity was much higher (78%) than that of melasolv. The inhibition of melanogenesis using Melan-A cell, the EtOAc fraction also showed higher inhibitory effect. The ethanol extract showed strong antioxidant activities, such as DPPH scavenging activity (IC50= 231.8 ㎍/㎖), NO scavenging activity (IC50= >1000㎍/㎖), superoxide scavenging activity (IC50= 23.6㎍/㎖), and xanthine oxidase inhibitory activity (IC50= 587.8㎍/㎖). Its EtOAc fraction showed the strongest antioxidant activities among several fractions. Also, the hexane and EtOAc fractions dose dependently inhibited the NO production in a RAW 264.7 cells. These results suggest that extract of Pimpinella komarovii could be used as functional biomaterial in developing a skin whitening agent having the antioxidant activity.

Most bioartificial liver (BAL) systems utilize porcine primary hepatocytes as a biological component of the systems. A packed-bed bioreactor with gel beads has higher surface area to volume ratio and cell capacity than those of a hollow fiber bioreactor. Therefore, the type of bioreactor may be a better alternative for hollow fiber-based BAL systems. In this study, we developed a simplified aseptic encapsulation process for hepatocyte spheroids using single vessel drop generator. All the processes of encapsulation, i.e. dropping, cross-linking, washing and size screening, were carried in the single vessel in which dropping multi-nozzle, washing media inlet and outlet port, double layered-meshes for size screening, and transfer port are installed with unique configuration. Large amount of hepatocyte spheroids were simply and safely immobilized with minimal damages by the single vessel drop generator and the micro-beads were transferred to packed-bed bioreactor system for a BAL system. This technology will offer a strict aseptic encapsulation process for various bio-processes that need micro-capsules.