Matrix Metalloproteinases (MMP)-2 and -9 are participated in embryo development, implantation, remodeling of epithelial cell and ovulation. The objective of this study is to evaluate an impact of MMP2 and MMP9 on embryonic developmental competence as well as gene expression profiles of in vitro-produced bovine embryos. After in vitro fertilization, embryos of all groups were transferred into IVC-2 medium treated with MMP2 and MMP9 to check the optimum concentration on the basis of embryo development competence and cell numbers. The optimum concentrations for MMP2 and 9 were 1,200 ng/ml and 300 ng/ml. The blastocyst development competence was not different among 1,200 ng/ml of MMP2 vs. 300 ng/ml of MMP9 vs. combined MMP2 + 9 vs. control groups (41.46 ± 10.66 vs. 37.73 ± 8.92 vs. 45.11 ± 11.41% vs. 41.59 ± 11.88, respectively). Furthermore, the developmental competences to hatching and hatched blastocysts were not also different among the same groups (79.84 ± 12.63 vs. 83.3 ± 17.46 vs. 78.55 ± 14.48% vs. 72.02 ± 14.09). In addition, total cell number was significantly (p<0.05) greater in blastocyst treated with MMP9 300 ng/ml among all treatment groups. On the other hand, there was no significant difference of ICM vs. TE ratio in all groups. The expression of five out of six genes (i.e., MMP2, MMP9, IFNt, SSLP1 and HNRNPA2B1) was different among the groups. The expression of IFNt and HNRNPA2B1 genes was significantly greater in MMP9 (p<0.05), but there was no difference of MMP9 expression between MMP2 and MMP9 group (p>0.05). The normalized expression of MMP2 and SSLP1 was greater in MMP2 than other groups (p<0.05). In conclusion, MMPs treatment during IVC-2 medium was remarkably effected on blastocyst developmental competence and gene expression profiles that are related to embryo quality and implantation.

Sargassum fusiforme has traditionally been widely consumed in Asia as a food, and it has gained much attention due to its high nutritional, pharmaceutical, and industrial value. This study aimed to examine the promotional effects of ethanol extract (ET) and fraction obtained from ethyl acetate (FR) of S. fusiforme on hair growth in C57BL/6 mice and HaCaT cells. Five-week-old mice were used to compare hair regrowth during application of ET and FR for 21 days. Hair regrowth was evaluated by macroscopic observation and verified by hematoxylin-eosin tissue staining. Levels of mRNA expression of factors relevant to the hair growth cycle such as keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), and transforming growth factor-beta1 (TGF-β1) were examined by quantitative polymerase chain reaction (qPCR). Our results showed that ET and FR successfully promoted hair regrowth in shaved C57BL/6 mice at a dose >20 mg/kg. Moreover, ET and FR were effective in stimulating expression of KGF and VEGF mRNAs in a dose-dependent manner, whereas TGF-β1 was not activated. These results indicate that ET and FR of S. fusiforme effectively promoted hair growth and gene expression relevant to hair growth cycles in both in vitro and in vivo models.

본 연구는 포화지방산과 불포화 지방산 첨가가 반추위 발효성상과 효소활력에 미치는 영향을 알아보기 위하여 Piromyces communis 곰팡이를 이용하여 in vitro 발효 시험을 실시하였다. 시험 Ⅰ과 Ⅱ의 대조구에는 지방산을 첨가하지 않았고, 처리구는 시험Ⅰ에서 palmitic acid, stearic acid, oleic acid를 첨가하였고, 실험 Ⅱ에서는 linoleic acid, linolenic acid, arachidonic acid를 첨가하였다. 두 실험 모두 39℃에서 7일간 (2, 3, 4, 6 및 7일) 실시되었다. 시험 Ⅰ의 가스발생량은 불포화 지방산인 oleic acid에서 가장 많이 생성되었고 H2가스의 비율은 가장 낮았다. 시험 Ⅱ의 linoleic acid 첨가구에서 2일에서 6일까지의 가스 발생량은 대조구에 비해 높았지만 이후에는 낮았다. 수소가스 생성비율은 linoleic acid 첨가구에서 가장 많았고 linolenic acid 첨가구에서 가장 낮았다. 또한 시험 Ⅰ의 pH는 6.63에서 5.85로, 시험 Ⅱ에서는 6.59에서 5.40으로 낮아졌다. Carboxymethylcellulase 활력은 발효시간(day) 경과에 따라 증가하는 경향이었고, 특히 arachidonic acid 첨가구의 증가폭이 컸다. 본 실험의 결과는 반추동물의 사료에 불포화지방산을 첨가하면 반추위곰팡이가 생성하는 수소를 불포화 지방산이 이용하여 메탄생성 억제에 도움이 될 것으로 기대된다.

한약추출 부산물의 영양가치 및 사료 가치를 평가하기 위해 in vitro 반추위 분해 시험을 하였다. 시험에 사용한 10가지 한약추출 부산물은 인삼(Ginseng radix alba), 백복령(Hoelen), 감초(Glycyrrhizaeradix), 당귀(Angelicae gigantis radix), 천궁(Cnidii rhizoma), 작약 (Paeoniae radix), 황기 (Astragali radix), 생강(Zingiberis rhizoma crudus), 대추(Ziziphi fructus), 진피(Aurantii nobilis pericarpium)로 한약재 중 빈번하게 사용하는 약재이다. In vitro 시험을 위해 50 mL serum bottle에 혐기상태로 McDougall's buffer와 반추위액을 2:1비율로 15 mL을 넣었다. 한약추출 부산물은 timothy(300 mg)를 기질로 3%, 5% 수준으로 첨가하였다. Serum bottle은 배양 시간대별(6, 12, 24, 48, 및 72 h)로 39℃에서 120 rpm으로 진탕 배양하였다. 총 가스 발생량은 한약추출 부산물 3% 첨가구 에서는 발효 24시간대에서 대조구가 생강(Zingiberis rhizoma crudus)구에 비해 많았으며, 발효 72시간 대에서도 대조구가 가장 많았고, 진피(Aurantii nobilis pericarpium)구에서 적었다(P<0.05). 한약추출부산물 5% 첨가구에서는 48시간대 가스 발생량이 진피(Aurantii nobilis pericarpium)구보다 대조구에서 많았으며, 72시간 발효구에서는 황기(Astragali radix)구에서 가장 적었다(P<0.05). 미생물 성장률은 한약추출 부산물 5% 첨가구의 48시간대를 제외한 모든 배양에서 유의적으로 증가하였다(P<0.05). VFA 농도는 배양시간의 따라 증가되는 경향을 보였으며, 배양 초기(6 h)에서 한약추출 부산물의 총 VFA 농도는 일부 한약추출 부산물 처리(작약, 황기, 진피)를 제외하고 유의적으로 증가하였다(P<0.05). 위의 결과로 한약추출 부산물은 미생물 성장률 및 발효성상 개선을 증진시킬 수 있으며, 결론적으로 조사료에 대한 한약추출 부산물 5% 대체가 가능할 것으로 예상된다.

한약추출 부산물의 영양가치 및 사료 가치를 평가하기 위해 in vitro 소화율 및 in situ 반추위 분해 시험을 하였다. 시험에 사용한 10가지 한약추출 부산물은 인삼(Ginseng radix alba), 백복령(Hoelen), 감초(Glycyrrhizae radix), 당귀(Angelicae gigantis radix), 천궁(Cnidii rhizoma), 작약(Paeoniaeradix), 황기(Astragali radix), 생강(Zingiberis rhizoma crudus), 대추(Ziziphi fructus), 진피(Aurantii nobilis pericarpium)로 한약재 중 빈번하게 사용하는 약재이다. 반추위 누관이 장착된 체중이 평균 620kg인 홀스타인 2두를 이용하여 in vitro 소화율 및 in situ 건물 소실률을 평가하였다. 건물소화율은 한약추출 부산물 3%, 5% 첨가에 따라 유의적으로 증가하였다(P<0.05). 펩신 소화율의 평균은 68.17%로 옥수수, 대두박보다 낮았다. 반추위 건물 소실률은 인삼(Ginseng radix alba), 당귀(Angelicae gigantis radix) 및 대추(Ziziphi fructus)가 유의적으로 높았으며(P<0.05), 백복령(Hoelen), 감초(Glycyrrhizae radix) 및 황기(Astragali radix)는 티모시 건초보다 유의적으로 낮았다(P<0.05). 한약추출 부산물의 12시간과 24시간대 건물 소실률 평균은 38.10, 44.43%이며, 티모시 건초의 평균 건물 소실률인 36.60, 43,25%보다 높았다. 이는 시험에서 사용된 한약추출 부산물은 한약재를 열수 추출 후 사용된 재료로써 한약추출 부산물의 물리적 처리에 의한 영향으로 건물 소실률이 티모시 보다 높은 결과를 보인 것으로 생각된다. 한약추출 부산물의 유효분해도는 인삼(Ginseng radix alba), 천궁(Cnidiirhizoma), 당귀(Angelicae gigantis radix), 대추(Ziziphi fructus) 및 진피(Aurantii nobilispericarpium)가 높은 추정치를 나타냈다. 결론적으로 한약추출 부산물은 반추동물용 조사료 대체자원으로 이용 가능 할 것으로 생각된다.

근육의 줄기세포라고 알려진 근육위성세포(satellite cells, SCs)는 근육의 성장과 발달 및 구조적, 대사적 형질 형성에 중요한 역할을 한다. 본 연구는 근육에서 추출한 SCs의 in vitro분화 성상과 형질을 분석함으로써 SCs가 근육의 성장 및 재생시 형질 형성에 미치는 영향을 구명하기 위해 수행되었다. 6주령 자돈의 Semitendinosus(ST) 근육의 적색근(red semitendinosus; RST)과 백색근(whitesemitendinosus; WST)에서 SCs를 추출한 후 in vitro 상에서 배양하여 SCs의 증식율, 분화율, 근육 구조단백질인 myosin heavy chain (MyHC) isoforms의 발현 성상을 분석하였다. 추출된 SCs의 숫자는 WST(63 ± 2 × 103 cells/g)보다 RST(79 ± 2 × 103 cells/g)에서 높게 (P<0.01) 나타났다. Celldoubling time은 RST에서 추출된 SCs가 WST보다 낮았고, 분화도 빨라 WST보다 먼저 근섬유를 형성하였으나, 분화 24h 후에는 WST 유래 SCs의 분화율이 RST보다 높게 나타났다(P<0.05). 하지만, 분화48h 후 RST와 WST의 SCs간 분화율은 차이가 없는 것으로 나타났다. MyHC type I isoform의 발현은 RST 유래 SCs에서 WST에 비해 높았으나 (P<0.01), type II isoform은 WST 유래 SCs에서 높게 나타났다 (P<0.05). 이 결과는 in vitro에서 배양된 SCs 에서 분화한 근섬유의 성장패턴 및 형질이 유래 근육의 형질을 그대로 반영한다는 것을 보여주는데, 이는 근육 내에 존재하는 SCs는 유래 근육의 형질을 기억하고 있거나, 특정 형질로만 분화되도록 정해져 있다는 것을 의미한다.

This study investigated the antibacterial effects of GR ethanol extracts (GRE), sodium chlorate (SC) and a combination of GRE and SC (GS) on Brucella abortus (B. abortus). The antibacterial activities of GRE, SC and GS towards B. abortus were evaluated by incubating B. abortus with GRE, SC and GS. Following treatment with GRE, SC and GS, B. abortus survival and intracellular proliferation in macrophages were monitored. In the cellular cytotoxicity assay, GRE, SC and GS are not cytotoxic at concentrations less than 400 μg/ml, 15 mM and 0.6GS (1 of GS, GRE 1,000 μg/ml + SC 30 mM), respectively. The viability of B. abortus was markedly decreased in a dosedependent manner in all treatment groups. In addition, B. abortus intracellular proliferation within macrophages was significantly reduced in cells treated with GRE (400 μg/mL), SC (15 mM) and 0.5GS (GRE 500 μg/mL + SC 15 mM) after 48 hr-incubation (GRE, p < 0.01; SC and 0.5GS, p < 0.001). Especially, in the treatment of GS, the synergistic effect of GRE and SC treatment on B. abortus in macrophage was observed. In conclusion, GS is useful as an antibacterial candidate against B. abortus, and can be applied in the field of meat and milk hygiene.

In porcine embryo culture, one of reactive oxygen species (ROS) is harmful factors that are made during in vitro culture. To decrease the detrimental effect of ROS on embryo development, superoxide dismutase, catalase and glutathione peroxidase could be used in the embryo culture. Out of these antioxidants, 7,8-dihydroxyflavone (7,8-DHF) was reported its antioxidant effects to prevent the glutamine-triggered apoptosis. Therefore, this study was performed to investigate the most appropriate concentration of 7,8-DHF in porcine embryonic development. For that, 5 different concentration (0, 0.1, 0.5, 1, 2 uM) of 7,8-DHF was supplemented in the porcine IVM media and then maturation and blastocyst formation rates were compared among 5 groups. In maturation rates of porcine oocytes, significant higher maturation rates was shown in the 1.0 uM group compared with another 4 groups (83.3 ± 2.1 vs. 80.7 ± 1.4, 79.8 ± 1.4, 78.3 ± 1.2, 79.4 ± 1.6), respectively (P<0.05). In the embryo culture, 1.0 uM group also showed the significant higher cleavage rates (76.8 ± 3.1 vs. 62.1 ± 5.0, 65.7 ± 4.0, 68.6 ± 3.7, 64.6 ± 4.0%) and blastocyst formation rates - (39.6 ± 4.0% vs. 28.6 ± 3.3, 31.1 ± 3.9, 29.3 ± 2.5, 39.6 ± 4.0, 26.4 ± 3.2%), respectively (P<0.05). There was no significant difference among 5 groups in the cell number of blastocyst (P<0.05). In conclusion, supplement of 1.0 uM of 7,8-DHF was effective to increase the porcine embryonic development competence as antioxidant to ROS.

The main purpose of this study is to estimate the effect of adding Tea-N-Tris (TES) to the freezing buffer for miniature pig sperm. In particular, we attempted to identify the association between the MMPs expression and the fertility and viability of frozen sperm from each extender (LEY (Lactose Egg-Yolk), TLE (TES + LEY), TFGE (TES + Fructose + Glucose Egg-Yolk)). In accordance with this, Hypoosmotic Swelling Test (HOST) respond test was the lowest among sperms frozen in LEY while the highest HOST respond was observed among sperms frozen in TLE. Furthermore, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of MMP-9 and MMP-2 was the highest in sperms frozen in LEY, Meanwhile, sperms from the TFGE and TLE group showed lower level of MMP-9 and MMP-2 expression in the order of TLE being the lowest. LEY group showed lower rate of blastocyst development than the TES supplement group, although the difference was not statistically significant. Meanwhile the rate of blastocyst development appeared similar when sperms from TLE and TFGE group were used for IVF. Together, these results indicate that adding Tea-N-Tris to the sperm freezing buffer only suppresses MMPs protein activation but also maximize in-vitro fertility, providing a means to improve the success rate in the in vitro manipulation of miniature pig sperm.

돼지 수정란의 체외 생산 효율성 향상을 위해서는 배발생율과 더불어 고품질의 배를 조기에 선별해야 한다. 체외 배 발생율에 대한 보고는 많지만, 고품질의 배를 선별할 수 있는 기술에 대한 연구는 거의 없었다. 본 연구에서는 돼지 난포란 유래 수정란의 체외배양에 있어서 배반포로의 배 발달과 생존에 미치는 Vitamin K1(vit K1) 첨가 농도, 시기 및 시간의 효과를 검토하였다. 1.0 μM, 3.0 μM 및 6.0 μM vit K1을 배양 1일째 24시간 첨가한 결과, 배반포 발달율이 시험군이 14.5 ± 4.3, 0.0 및 0.0%로써 대조군의 35.5 ± 3.2%에 비하여 유의하게 낮았고(p<0.05), 배반포의 생존율도 대조군이 31.8 ± 2.6%로써 시험군의 22.2 ± 2.9, 0.0 및 0.0%에 비하여 유의하게 높았다(p<0.05). 상기 첨가 농도에서 첨가 시간을 달리한 결과, 1.0 μM 농도에서 6시간 처리군의 배반포 발달율과 생존율이 각각 26.5 ± 2.9% 및 47.2 ± 2.8%로써 가장 높았고 특히, 12시간 처리군보다 유의하게 높았다(p<0.05). 3.0 μM 농도에서는 대조군의 배발달율이 36.4 ± 3.1%로 가장 높았으나, 생존율은 3.0시간 첨가군이 41.7 ± 3.2%로 대조군에 비하여 유의하게 높았다(p<0.05). 6.0 μM 농도에서도 배발달율은 대조군(32.0 ± 2.8%), 생존율은 0.5시간 첨가군(42.9 ± 1.8%)이 가장 높았다. 각각의 vit K1 첨가 농도와 시간을 기준으로 서로 다른 배양 시기에 첨가한 결과, 1.0 μM 6시간 첨가군에서는 배반포 발달율은 배양 4일째 첨가군, 생존율은 배양 2일째 첨가군이 가장 높았다. 한편, 3.0 μM 3.0시간 및 6.0 μM 0.5시간 첨가군에서는 배양 4일째 첨가군의 배반포 발달율(59.5 ± 4.1% 및 50.0 ± 3.6%)과 생존율(72.7 ± 5.4% 및 79.2 ± 4.0%)이 대조군과 다른 시험군에 비하여 유의하게 높았다(p<0.05). 한편, vit K1 첨가에 따른 배반포의 세포 수를 조사한 결과, 첨가군(1.0 μM 6시간 배양 2일째, 3.0 μM 3.0시간 배양 4일째 및 6.0 μM 0.5시간 배양 6일째)이 53.4 ± 5.8, 49.4 ± 3.8 및 51.5 ± 4.5개로써 대조군의 40.2 ± 2.3개에 비하여 유의하게 많았다(p<0.05). 그러나 사멸세포 수는 시험군이 3.2 ± 0.9∼3.7 ± 2.1개로써 대조군의 4.2 ± 1.2개보다 적었으나, 유의차는 없었다. 세포 사멸 유도 유전자인 Bax mRNA 발현은 처리군과 대조군은 비슷하였으나, 세포 사멸 억제 유전자인 Bcl-xL mRNA 발현은 처리군이 대조군보다 높았고 특히, 6.0 μM 0.5시간 배양 4일째 첨가군이 가장 높았다. 이상의 결과로부터 돼지 미성숙 난포란 유래 수정란의 체외 배양에 vit K1의 첨가는 배반포의 생존율과 세포수 증가에 효과적이었다. 그 이유에 대해서는 아직 많은 부분이 밝혀져야 되겠지만, 고품질의 배반포 조기 선발에는 활용이 가능할 것으로 생각된다.

본 연구는 돼지 정자의 동결 건조 시 동결 보호제인 trehalose의 효과와 동결 건조 시간과 동결 건조 후 저장 기간에 따라 정자의 생존성과 체외 성숙 난자 내 동결 건조 정자를 직접 주입한 후 전핵 형성율, 난할율 그리고 배발달 성적을 조사하였다. 동결 건조 후 정자의 생존율은 trehalose 무첨가구에 비해 trehalose를 첨가한 처리구에서 높은 생존율을 보였으며, 75 mM의 trehalose를 첨가하여 동결 건조한 정자들의 생존율이 가장 높은 것으로 나타났다. 또한, 동결 건조 후 저장 기간이 길어질수록 생존율이 낮아지는 경향이었다. 체외 성숙 난자 내 동결 건조 정자를 직접 주입 후 전핵 형성율은 trehalose 첨가구에서 유의적으로 높았으며(p<0.05), 난할율과 배발달 성적도 trehalose 첨가구에서 유의적으로 높았고(p<0.05), 정자의 동결 건조 시간이 짧을수록 높은 난할율과 배발달율을 보였다.

Recent 2 decades, including in vitro maturation (IVM), assisted reproductive technologies (ARTs) achieved noteworthy development. However the efficiency of ARTs with in vitro matured oocytes is still lower than that with in vivo oocytes. To overcome those limitations, many researchers attempted to adapt co-culture system during IVM and consequently maturation efficiency has been increased. The beneficial effects of applying co-culture system is contemplated base on communication and interaction between various somatic cells and oocytes, achievement of paracrine factors, and spatial effects of extracellular matrix (ECM) from somatic cell surface. The understanding of co-culture system can provide some information to narrow the gap between in vitro and in vivo. Here we will review current studies about issues for understanding cu-culture system with various somatic cells to improve in vitro maturation microenvironment and provide bird view and strategies for further studies.

Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or 37℃. In case of ICR strain, primary TSCs cultured at 37℃ showed significantly higher proliferation and viability than those at 35℃ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the 35℃ culture condition. Moreover, sub-passage of primary TSCs at 35℃ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at 35℃, which showed no significant difference in the viability, compared to those at 37℃. Furthermore, sub-passaged TSCs cultured at 37℃ showed no significant differences in proliferation and viability, compared to those at 35℃. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at 35℃, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at 37℃. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.

In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phos-phodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, 67.57±4.11% aging, 44.61±6.4%) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of con-trol group intensity rate (51.53.±3.80), aging group (68.10±5.54) and treatment of caffeine (45.04±2.98). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging (90.44±10.18 VS 67.88±7.72). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro

In order to investigate the effects of antioxidants on the culture of mouse preantral follicles in vitro, we examined the effects of taurine, glutathione and catalase on their growth and maturation. Addition of taurine was not effective on the survival of preantral follicles. However, metaphase II rates of oocytes within preantral follicles were signifi-cantly higher in 1 mM treated group than in control and 10 mM treated group (p<0.05). Glutathione did not improved the rates of survival and metaphase II oocytes. However, metaphase II rates of oocytes progressively decreased with increasing glutathione concentration. Catalase also showed that the rates of survival and metaphase II oocytes pro-gressively decreased with increasing concentration. Especially, all of preantral follicles cultured in medium containing 100 IU/ml catalase were degenerated. These results suggest that low concentraion of taurine, as an antioxidant, have positive effect on the culture of mouse preantral follicles in vitro.

Human dental pulp stem cells (DPSCs) are multi-potent mesenchymal stem cells that have several differentiation potentials. An understanding of thetissues that differentiate from these cells can provide insights for future regenerative therapeutics and tissue engineering strategies. The mesiodens is the most frequent form of supernumerary tooth from which DPSCs can differentiate into several lineages similar to cells from normal deciduous teeth. Recently, it has been shown that nanoscale structures can affect stem cell differentiation. In our presentstudy, we investigated the effects of a 250-nm nanoscale ridge/groove pattern array on the osteogenic and adipogenic differentiation of dental pulp cells from mesiodenscontaining human DPSCs. To this end, the expression of lineage specific markers after differentiation induction was analyzed by lineage specific staining and RT-PCR. The nanoscale pattern arrayed surface showed apositive effect on the adipogenic differentiation of DPSCs. There was no difference between nanoscale pattern arrayed surface and conventional surface groups onosteogenic differentiation. In conclusion, the nanoscale ridge/groove pattern arrayed surface can be used to enhance the adipogenic differentiation of DPSCs derived from mesiodens. This finding provides an improved understanding of the effects of topography on cell differentiation as well as the potential use of supernumerary tooth in regenerative dental medicine.

귀리, 콩, 감자, 밀에서 유래한 다양한 식이섬유를 밀가루 대신 10-40%까지 대체함에 따른 수화능, 동적점탄성, 용매흡착능 및 in vitro starch digestion을 비교하였다. 귀리, 콩, 감자, 밀에서 유래한 다양한 식이섬유를 식이섬유의 대체비율이 증가할수록 수분흡착능과 팽윤력은 증가한 반면, 수분용해도는 감소하였다. 밀가루 대신 콩과 감자 식이섬유를 대체함에 따라 동적점탄성은 증가하였으나, 귀리와 밀 식이섬유 대체에서는 오히려 감소하는 경향을 보였다. 식이섬유 대체에 따른 밀가루의 용매흡착능 변화를 분석한 결과, 낮은 대체량의 귀리 식이섬유와 높은 대체량의 콩과 감자 식이섬유가 활용 가능성을 보였다. 원료별 식이섬유 대체 시 밀가루 겔의 전분소화 패턴은 식이섬유 대체비율이 증가함에 따라 glucose 방출은 감소하였으나, 원료별 특성에 따른 유의적인 차이는 없었다. 특히, 밀식이섬유를 제외한 귀리, 콩, 감자 식이섬유는 모두 RDS감소와 RS증가에 따른 pGI 저하를 보여 실제 식품소재로 활용 시 가공적성을 유지할 수 있다면 전분소화지연효과를 기대할 수 있음을 확인할 수 있었다. 한편, 식이섬유의 원료에 따른 식이섬유 조성과 전분소화 관련 특성 간 상관관계를 분석한 결과, 전분소화를 지연시키는 효과는 수용성 식이섬유보다는 불용성과 총식이섬유 함량이 중요한 인자임을 알 수 있었다.

Royal jelly (RJ) is exclusive food that is secreted from the hypopharyngeal and mandibular glands of worker honeybees, and it is well known to be a necessary for the growth of the queen honeybee Although fresh royal jelly have been demonstrated to enhance wound healing, the wound healing effects of water soluble royal jelly (WSRJ) have not been elucidated. We investigated whether WSRJ promotes the migration, attachment, and proliferation of human dermal fibroblasts (HDFs) during in vitro wound healing. HDFs were treated with 1-5ug/ml WSRJ and RJ for up to 24hr following wound formation. Cell migration was assessed by measuring recovery from wound margin, while cell attachment and proliferation were determined by MTT assay. By observing the numbers of cell attached, we confirmed that not only WSRJ but also RJ did not affect on the initial cell adhesion. WSRJ (5 ug/ml) enhanced cell migration rate approximately 84.3% in HDFs at 24hr, whereas RJ (5 ug/ml) increased cell migration rate 71.3% in HDFs at 24hr, which is similar to cell migration rate of WSRJ 1 ug/ml (73.7%). In cell proliferation assays, WSRJ induced an increase in the number of HDFs, compared with control and RJ. In conclusion, WSRJ promotes cell migration with increased cell proliferation in an in vitro wound healing model.