The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro−grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in “BaeYun No. 3” (55.7%) and “Whanggeum” (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro−grown plantlets of pear germplasm.

Background: Purple potato contain sufficient phenolic compound and flavonoid which has high antioxidant capacity. Due to poor water solubility of phenolic compounds and quick oxidation of anthocyanin, we could not get maximum health benefits from purple potato. Therefore, we developed surfactant based aqueous nano suspension to enhance the solubility of phenolic compounds and protect the oxidation of anthocyanin from purple potato. Methods and Results: Two types of surfactant were used in this experiment based on hydrophilic-lycophilic balance viz. Brij and Span. In our study, lycophilic surfactant showed highest efficiency in TP extraction compared to water and lipophilic surfactant below 10 mM concentration. On the other hand, lipophilic surfactant showed highest efficiency in extracting flavonoid content. Conclusion: It is concluded that hydrophilic surfactant was significantly increased phenolic compounds five times, and lipophilic surfactant increased flavonoid two times, and anthocyanin three times compared to control. Therefore, total antioxidant capacity was increases two times compared to control.

Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulationvitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at 25°C. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

This experiment was carried out to enhance plantlet conversion and ex vitro survival of encapsulated somatic embryos of Siberian ginseng. Cotyledonary somatic embryos were encapsulated with 3.0% sodium alginate and 96% of conversion rate in terms of plantlet with well-developed epicotyl marked when the encapsulated embryos were placed on perlite soils wetted with sucrose solution as for carbon source. However, post-germinative growth of encapsulated embryos was suppressed in case of sucrose did not added. Instead of sucrose alone, the addition of both sucrose and starch to the sodium alginate enhanced the post-germinative growth of the embryos. In sodium alginate matrix with 2% sucrose, the survival rate of the encapsulated embryos was more than twice (23.5%) that of ones without sucrose (10.0%). Embryos encapsulated with both 2% sucrose and 1% starch showed the highest percentage (42.1%) of survival rate was shown. In analysis of Iodine staining and starch content in the sodium alginate matrix, the starch component was decomposed when the embryos started to germinte. This result indicated that the carbohydrate treatments (starch and sucrose) in the encapsulation matrix enhanced the survival rate of post-germinative growth of encapsulated embryos in Siberian ginseng.

새로운 베지클인 glyceryl citrate/ lactate/ linoleate/ oleate를 이용한 수중유형 형태의 아스타잔틴 나노에멀젼에 대해 항산화 효과, 세포 생존력, 단백질과 관련한 효소의 영향, 피부 침투도 그리고 피부에 대한 보습 및 탄력 등의 약용화장품적인 측면에서의 전반적 연구를 실시하였다. 항산화력 및 세포 생존력에 대해선 각각 DPPH법과 MMT assay를 이용하여 측정하였다. 아스타잔틴 나노에멀젼에 대한 또 다른 성질은 2D-Page를 이용한 단백질 분석 및 컨포칼, in-vivo 테스트를 통해 측정하였다. 본 연구를 통해, 아스타잔틴을 포함하는 나노에멀젼은 MMP발현에 관련한 단백질 억제 및 세포외 기질의 분해를 막고 라디칼의 소거에 매우 우수한 결과를 보였다. 종전의 레시친을 이용한 나노에멀젼 보다는 새로운 베지클을 이용한 아스타잔틴 나노에멀젼의 피부 침투가 매우 효과적임을 CLSM을 통해 측정하였다. 또한 28일 동안의 한국 성인 여성 11명을 통한 보습 및 탄력 인비보 테스트에서 우수한 효과를 확인할 수 있었다.

아스타잔틴을 포함하는 수중유형 형태의 나노에멀젼을 고압균질기를 이용하여 제조하였다. 이에 유화조건, 유화제 종류, 유화제 농도 그리고 아스타잔틴 농도에 따라 최적화를 되었다. 나노에멀젼의 안정성은 제타포텐셜, FF-SEM, 입도분석기, 색차계를 이용하여 측정하였다. 제조된 아스타잔틴 나노에멀젼의 입도는 160 ~190 nm로 균일하였으며 레시친에 의한 나노에멀젼 보다 glyceryl citrate/lactate/linoleate/oleate에 의한 나노에멀젼이 더욱 안정하고 균일한 입도분포를 가졌다. 아스타잔틴의 봉입도는 HPLC, FF-SEM을 이용하여 확인 하였으며, 제형 구성 후 보관조건에서의 안정도 및 제타 포텐셜 값도 -41의 우수한 결과를 나타내었다.

본 연구는 레시틴으로 나노입자화 시킨 티아민 디라우릴 설페이트의 향장활성 증진에 관한 것이다. TDS를 포집시킨 나노입자는 150 ~ 200 nm의 크기를 나타내는 구형이며, 또한 제타포텐셜을 측정하여 여러 pH범위에서 안정한 것을 확인하였다. TDS 나노입자는 인간 섬유아세포(CCD-986sk)에 높은 농도를 처리하여도85%의 세포생존률을 보였다. 자유라디칼소거활성 실험을 진행한 결과 나노입자화하지 않은 TDS 희석액(1.0mg/mL)은 81.6%의 활성을 나타내었고, 나노입자화한 TDS 용액은 이보다 더 높은 88.1%의 높은 라디칼 소거활성을 보였다. TDS 나노입자는 자외선을 조사시킨 CCD-986sk에서 MMP-1의 발현을 41.4% 감소시켰다.TDS 용액과 TDS 나노입자를 가지고 salmonella typhimurium, listeria monocytogenes에 대하여 항균활성을 측정하였다. TDS 나노입자의 경우 양성대조군의 항균활성과 비슷한 결과를 나타내었다. 이러한 결과들로TDS 나노입자가 항산화, 미백, 주름개선 효능같은 향장 소재로서의 적용이 가능할 것이라 생각된다.

In this study, we investigated antioxidant activities and whitening effects of Acer mono sap by encapsulation of nanoparticles. Acer mono sap was through ultra high pressure process and then encapsulated by lecithin. Nano-encapsulated The nanoparticles of Acer mono sap showed highest free radical scavengering effect as 89.7% in adding sample (1.0 mg/ml), compared to sap of non-encapsulation. It was showed strong inhibition effect on melanin production test by Clone M-3 cells as 47.8%. High inhibitory of tyrosinase was also measured as 85.8% by adding lecithin nano-particle of 1.0 mg/ml. The nano-particles also showed 14.8% of low cytotoxicity against human normal fibroblast cells in adding 1.0 mg/ml of the highest concentration. These results indicate that Acer mono sap may be a source of cosmetic agents capable of improving whitening effect and antioxidant activites.

This study showed the increase of antitumor activities of water soluble E. sinica extract by nano-encapsulation process with lecithin. Five groups of lecithin only group (LO), lecithin nano-encapsulated E. sinica group (LE), E. sinica only group (EO), one negative control group (NCO) and positive control group (PCO) were set for several anticancer experiment and fed into Sarcoma-180 injected mice. The cytotoxicity of LE on the human normal kidney cell (HEK293) showed 14.8% lower than 19.2% of EO and 18.4% of LO. Growth of human liver carcinoma cell and human stomach carcinoma cell as representative of digestive system in vitro was inhibited up to about 85.1% and 87.3%, in adding 1.0 mg/ml of LE, which values 15% higher than that from conventional EO. The survival rates of each mice group were 40%, 63%, 48%, 33% and 100%, respectively after 40 days of injecting Sarcoma-180. The increment of their body weights of the extract feeding groups was suppressed down to 10~15%, compared to the negative control. The nano-particles also reduced the hypertrophy of the internal organs such as spleen and liver down to 15~20%, compared to those as the other groups. Among them, LE effectively reduced the size of tumor form to 20%. From these results, in vitro and in vivo antitumor activities of E. sinica could be enhanced by using nano-encapsulation process with lecithin because of better permeation into the cancer cells by confocal observations.

Phosphatidylcholine was used to encapsulate aqueous extracts of Centella asiatica, and its biological activity was compared with another aqueous extracts. Nanoparticle of C. asiatica was made by encapsulation to w/o type spherical liposome which of aqueous extracts seized with oil phase as 78.2 nm average diameter. Cytotoxicity of the nanoparticle was measured on human skin fibroblast cells, CCD-986sk, and showed lower cytotoxicity on 1.0 mg/ml of highest concentration as 28% than that of another extracts. The nanoparticle showed the highest promotion of human B and T cell growth up to 138% and 135%, respectively, compared to the control. and the NK cell growth was promoted up to 8% higher than the control in proportion to secretion of IL-6 and TNF-α from immune cell growth. Also nanoparticle showed highest inhibition activity of hyaluronidase on 1.0 mg/ml of highest concentration as 60.5%. It seems that because of enhanced biological application of aqueous extracts on cell through nano-encapsulation process.

This study was performed to improve immune activities of Rubus coreanus Miquel by encapsulation of nanoparticles. Immuno-activities of R. coreanus were investigated through aqueous extracts associated with process of water at 60℃. It showed high promotion of human B and T cells growth about 50%, compared to the case of other conditions. The secretion of IL-6 and TNF-α was also enhanced as 2.44×10-4pg/cell and 1.94×10-4pg/cell, results by adding nano samples. NK cell activation was improved up to 29% higher than the conventional extraction process. The secretion of NO from macrophage showed 14.9 μm on the nano-encapsulation process extracts, which was higher than others. The size of nanoparticles was in the range of 50~300 nm, which can effect the penetration into the cells. It was clearly observed by real time confocal microscope.

This study was performed to investigate improving immune activities of natural water-soluble sulforaphane extracted from Brassica oleracea var. italica by nano encapsulation process. The nanoparticles of the sulforaphane extracted with ultrasonification process at 60℃ promoted human B and T cell growth, about 7~35% compared to the control. The secretion of IL-6 and TNF-α from T cells were also enhanced as 2.6×10-4pg/cell and 2.1×10-4 pg/cell, respectively, by the adding nano samples. NK cell activation was improved about 8%, compare to the control in adding cultured medium of T cell added nano samples. It was also found that sulforaphane extracted from B. oleracea var. italica had highly inhibitory activity on hyaluronidase as IC50 about 200 μg/ml. It can be concluded that natural water-soluble sulforaphane samples by nano-encapsulation, each size is 200 nm, extracted from B. oleracea var. italica has high immune activities through higher efficiency of bio-activation than conventional extracts.

본 연구는 유용성 물질 포집에 효과적인 solid lipid nanoparticle (SLN)을 이용하여 수용성물질인 LA-PEG을 새로운 제조방법에 응용하여 실험하였다. 지질로 사용된 오일은 coconut oil, macadamia oil, 그리고 jojoba oil 3가지로 이들의 특징은 생분해성이 강하다. 외부유화제로는 Tween 20, Tween 60을 이용하여 T-SLN을 제조하였으며, SLN의 입자 분포를 비교 분석한 결과 coconut oil을 지질로 하여 사용한 것이 크기가 가장 작았으며 사용한 계면활성제의 양에 따라 입자크기와 분포형태가 달라졌다. 1 %의 Tween 60과 macadamia oil을 이용한 베이스가 입자크기가 가장 컸다. 방출관찰결과 coconut oil을 지질로 한 2 %의 Tween 20의 베이스가 가장 늦게 방출되었고, 가장 빠른 방출한 것은 Tween 60 2 % 베이스였다.

A simplified technique which cryoprotects zygotic embryos by encapsulation-dehydration was developed for the germplasm conservation of herbaceous peony (Paeonia lactiflora Pall.). The highest survival rate (85~%) was obtained from embryos treated by encapsulation-dehydration. The zygotic embryos were precultured on MS medium containing 0.3mg/L GA3 for 1 day. The precultured embryos were encapsulated in 3~% (w/v) alginate beads and immersed for 1 h in MS medium containing 2 M glycerol and 0.5 M sucrose. The encapsulated embryos were dehydrated for 5h by air drying prior to direct immersion in liquid nitrogen. This encapsulation-dehydration method appears to be a promising technique for germplasm cryopreservation of a herbaceous peony.

The rol e of osm otol erance and dehydr ation, in terms of both medium com posi tion and exposur e duration, on sur vi val ofem bryos of Citrus m adurensi s after cryopr eser vation usi ng the encapsul ation-dehydr ation and modified encapsul ation-dehydr

현탁배양으로 체세포배를 대량 생산하는데 관여하는 요인 들을 조사분석하고. 생산된 체세포배를 encapsulation시켜 인공종자화가 가능한지를 검토한 결과를 요약하면 아래와 같다. 1. 현탁배양에서 LS(Linsmeier-Skoog) 배지가 어뢰형과 자엽기의 체세포배 형성율이 높았다. 2. 현탁배양시 체세포배 형성에 영향하는 탄소원은 sucrose가 가장 효과적이었다. Gtucose에서도 자엽 기의 체세포배가 형성되었으나 sucrose에서보다는 적었다. Fructose, lactose 및 maltose는 효과가 없었다. 3. 현탁 LS 배지에서 sucrose 농도는 3%가 체세포배 형성에 적합한 농도로 나타났다. 4. 현탁 LS 배지에서 질소원을 달리하여 체세포배 형성율을 조사한 결과 암모니아태 질소의 영향은 거의 없는 것으로 나타났다. 또한 암모니아태 질소와 질산태 질소의 비율을 413(mg/l) : 1900(mg/l)로 하는 것이 바람직하다고 판단되었다. 5. 현탁배양에서 계대배양 3회까지는 체세포배형성에 큰 차이가 없는 것으로 나타났다. 그러나 불필요하게 계대배양 회수를 증가시킬 필요는 없다고 본다. 6. 고형배지에서 체세포배 형성을과 형성 수에는 BA단독보다는 NAA를 혼용하는 것이 효과적이었으며 이 효과는 BA 농도가 높은 경우에 크게 나타났다. 7. 고형배지에서 체세포배 형성율과 형성 수에는 kinetin 단독보다는 NAA와 혼용하는 것이 바람직하며 이 효과는 kinetin의 농도가 높은 경우가 좋았다. 8. 고형배지에서 체세포배 형성율과 형성 수에는polyamines 중에서 spermidine과 spermine이 효과적이었으며 spermine 10mg/l처리에서 가장 많은 체세포배가 형성되었다. 9. 기내묘에서 발생하는 vitrification을 억제시키기 위하여 질산은을 처리하더라도 체세포배 형성이나 형성율에 별다른 영향이 없었다. 10. 체세포배를 encapsulation 할 때 인공종자의 구형 형성은 sodium alginate 농도를 3%로 인한 경우 가장 좋았으나 인공종자의 발아율은 2.5%에서 100% 발아가 되었고 3%에서는 88%가 발아하였다. 11. 인공종자는 발아하여 정상적으로 생장하면서 shoot와 root를 형성하였다.