• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권2호
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• pp.87-93
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• 2010

Introduction: In our previous studies, we isolated porcine skin-derived mesenchymal stem cells (pSDMSCs) from the ears of adult miniature pigs and evaluated the pluripotency of these pSDMSCs based on expressions of transcription factors, such as Oct-4, Sox-2, and Nanog. Moreover, the characteristic of mesenchymal stem cells was revealed by the expression of various mesenchymal stem cell markers, including CD29, CD44, CD90, and vimentin. The aim of this study was to evaluate in vivo osteogenesis after maxillary sinus lift procedures with autogenous pSDMSCs and scaffold. Materials and Methods: The autogenous pSDMSCs were isolated from the 4 miniature pigs, and cultured to 3rd passage with same methods of our previous studies. After cell membranes were labeled using a PKH26, $1{\times}10^{7}$ cells/$100{\mu}L$ of autogenous pSDMSCs were grafted into the maxillary sinus with a demineralized bone matrix (DBM) and fibrin glue scaffold. In the contralateral control side, only a scaffold was grafted, without SDMSCs. After two animals each were euthanized at 2 and 4 weeks after grafting, the in vivo osteogenesis was evaluated with histolomorphometric and osteocalcin immunohistochemical studies. Results: In vivo PKH26 expression was detected in all specimens at 2 and 4 weeks after grafting. Trabecular bone formation and osteocalcin expression were more pronounced around the grafted materials in the autogenous pSDMSCs-grafted group compared to the control group. Newly generated bone was observed growing from the periphery to the center of the grafted material. Conclusion: The results of the present study suggest that autogenous skin-derived mesenchymal stem cells grafting with a DBM and fibrin glue scaffold can be a predictable method in the maxillary sinus floor elevation technique for implant surgery.

• 한국수정란이식학회지
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• 제32권3호
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• pp.209-220
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• 2017

The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of $17{\beta}$-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor ${\alpha}$ ($ER{\alpha}$) and $ER{\beta}$ in NF-pBMSCs, but NM-pBMSCs were only correlated with $ER{\alpha}$. Moreover, E2-treated NF-pBMSCs decreased in ${\beta}$-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.

• Animal Bioscience
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• 제34권4호
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• pp.546-557
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• 2021

Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

• 생명과학회지
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• 제17권10호
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• pp.1315-1320
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• 2007

미니돼지의 긴 뼈의 뼈끝(epiphysis) 성장에 대한 연구를 위해 상완골, 요골, 척골, 대퇴골, 경골의 근위 및 원위 뼈끝 부위를 중심으로 방사선 촬영을 실시하였다. 실험에 이용된 미니돼지의 연령대는 4주, 8주, 12주, 20주, 40주, 48주, 96주, 144주령으로 총 58마리였다. 뼈 성장과정에 대한 평가는 Owada & Sutow가 제안한 11가지 기준을 토대로 하였다. 긴 뼈에서 이차뼈발생중심은 4주령에서 40주령까지 빠른 성장을 보였고, 그 이후 96주령까지는 초기보다 느린 성장을 보였다. 가장 빠른 뼈끝 유합시기는 96주령으로 근위 요골, 근위 및 원위 대퇴골에서 관찰되었다. 미니돼지의 일반적인 긴 뼈의 완전한 뼈끝 유합은 144주령에 나타났으며 근위 상완골, 근위 및 원위 척골, 원위 요골, 그리고 근위 경골에서 관찰되었다.

• Journal of Periodontal and Implant Science
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• 제51권3호
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• pp.179-188
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• 2021

Purpose: Due to the difficulty of the hygienic care and sanitary management of abutment teeth and subpontic areas associated with fixed dental prostheses (FDPs), intrabony defects occur and accelerate due to the accumulation of plaque and calculus. This study aimed to evaluate the efficacy of regenerative periodontal surgery for intrabony defects associated with FDPs. Methods: The study inclusion criteria were met by 60 patients who underwent regenerative treatment between 2016 and 2018, involving a total of 82 intrabony defects associated with FDPs. Periodontal osseous lesions were classified as 1-, 2-, and 3-wall intrabony defects and were treated with an enamel matrix derivative in combination with bone graft material. The changes in clinical (pocket probing depth [PPD] and clinical attachment level [CAL]) and radiographic (defect depth and width) outcomes were measured at baseline and at 6, 12, and 24 months. Results: Six months after regenerative treatment, a significant reduction was observed in the PPD of 1-wall (P<0.001), 2-wall (P<0.001), and 3-wall (P<0.001) defects, as well as a significant reduction in the CAL of 2-wall (P<0.001) and 3-wall (P<0.001) intrabony defects. However, there was a significant increase in the CAL of 1-wall intrabony defects (P=0.003). Radiographically, a significant reduction in the depth of the 3-wall (P<0.001) defects and a significant reduction in the width of 2-wall (P=0.008) and 3-wall (P<0.001) defects were observed. The depth decreased in 1-wall defects; however, this change was not statistically significant (P=0.066). Conclusions: Within the limitations of the current study, regenerative treatment of 2- and 3-wall intrabony defects associated with FDPs improved clinical and radiological outcomes. Additional prospective studies are necessary to confirm our findings and to assess long-term outcomes.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.157-162
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• 2006

본 연구에서는 인간 부자상선 호르몬의 발현을 유도적으로 조절할 수 있는 retrocvirus vector system을 확립하고자 하였다. 이에 tetracycline계 물질로 발현을 유도적으로 조절할 수 있는 one vector 형태의 Tet-On system을 이용하였으며 WPRE 서열을 도입하여 유도적 조건에서 외래 유전자의 발현을 증가시켰다. 구축한 각각의 표적세포에서 RT-PCR과 ELISA를 이용하여 hPTH유전자의 발현 정도를 비교 측정한 결과, WPRE가 hPTH의 3' 위치에 도입된 $RevTRE-PTH-WPRE-CMVp-rtTA2^sM2$ virus를 이용하여 유전자를 전이시킨 경우에 hPTH의 발현량이 가장 높은 것으로 나타났고, 또한 유도율도 가장 큰 것으로 확인되었다. 이 system을 이용하여 생산한 고감염가의 virus는 인간의 부갑상선 호르몬을 생산하기 위한 동물세포주의 구축이나 형질전환 동물의 생산에 있어서 매우 효율적인 유전자 전이 수단이 될 것으로 사료된다.

• 대한정형외과스포츠의학회지
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• 제8권1호
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• pp.26-32
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• 2009

목적: 사체의 원위 대퇴골을 이용하여 각각의 인대가 붙는 부위의 골 강도를 비교함으로서 재건술이나 인대 보강 술식시 고정에 참고할 수 있는 지표로 삼고자 하였다. 대상 및 방법: 10구의 사체, 15개의 원위 대퇴골을 이용하여 골밀도 측정 후 5 개의 원위 대퇴골에 5.0 mm 유관나사를 각각의 인대 부착부, 즉 전방 십자 인대, 후방 십자 인대, 내측 측부 인대, 외측 측부 인대의 부착부에 삽입하고 최대 인장 강도를 측정하였고(실험 1), 10개의 원위 대퇴골을 이용하여 돼지의 신전건을 각각의 인대 부착부, 즉 전방 십자 인대, 후방 십자 인대, 내측 측부 인대, 외측 측부 인대의 부착부에 삽입하고 흡수성 간섭 나사로 고정 후 최대 인장 강도를 측정하였다(실험 2). 결과: 골밀도 검사는 실험 1이 $1.205{\pm}0.137\;g/cm^2$, 실험 2가 평균 $1.236{\pm}0.089\;g/cm^2$로 양 군간의 차이는 없었으며, 실험 1의 최대 인장 강도는 전방 십자 인대 군이 평균 $519.1{\pm}111.7$ N, 후방 십자 인대 군이 평균 $638.9{\pm}144.4$ N, 내측 측부 인대군이 평균 $169.7{\pm}56.0$ N, 외측 측부 인대 군이 평균 $225.6{\pm}61.5$ N으로 후방 십자 인대 군, 전방 십자 인대 군, 외측 측부 인대 군, 내측 측부 인대 군 순이었고, 실험 2의 최대 인장 강도는 전방 십자 인대 군이 평균 $310.6{\pm}31.0$ N, 후방 십자 인대 군이 평균 $379.9{\pm}47.4$ N, 내측 측부 인대 군이 평균 $104.01{\pm}14.4$ N, 외측 측부 인대 군이 평균 $131.5{\pm}21.9$ N으로 실험 1과 동일한 순이었다. 양 실험 모두 전방 십자 인대 군과 후방 십자 인대 군 간, 내측 측부 인대 군과 외측 측부 인대 군 간 강도의 유의한 차이는 없었으며, 내측 측부 인대 군과 외측 측부 인대 군이 전방 십자 인대 군과 후방 십자 인대 군에 비해 강도가 유의하게 낮았다. 결론: 대퇴 터널과 동일한 두께, 최소 길이의 간섭 나사못 고정은 불충분한 고정력을 제공하므로 전방 십자인대, 후방 십자인대, 내, 외측 측부 인대 재건술시 부가적인 술식이 필요하며 특히 외측 측부 인대와 내측 측부 인대 부착 부위의 강도는 후방십자 인대와 전방 십자 인대 부착 부위의 강도에 비해 현저히 약하므로 외측 또는 내측 측부 인대 재건술 및 보강술시 고정 방법에 대해 주의를 요하며 이를 보강할 수 있는 술식이 필요로 하다고 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권7호
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• pp.925-937
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• 2016

In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

• 한국가금학회지
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• 제31권4호
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• pp.245-253
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• 2004

본 연구는 육계사료 내 rPST-yeast culture의 첨가 급여가 육계의 성장성적, 도체 특성 및 골격발달 등에 미치는 영향을 규명하기 위해 수행하였다. 2일령의 Ross 육용종 수평아리 460수를 공시하여 항생제, rPST-yeast culture 및 yeast culture가 첨가되지 않은 대조구 사료와 대조구 사료에 항생제(chlorotetracycline)만을 $0.1\%$ 수준으로 첨가한 실험사료(T1), rPST-yeast culture를 각각 $0.1\%$ 및 $0.2\%$ 수준으로 첨가한 실험사료(T2 및 T3) 및 yeast culture만을 $0.2\%$수준으로 첨가한 실험사료(T4)를 각각 6주간 급여하였다. 사료섭취량과 증체량은 주 단위로 조사하였다. 실험 5주차 종료 시에 각 처리구별로 10수씩 선발하여 도살하였고, 간, 비장, 복강지방 및 가슴근육의 상대적 중량과 가슴근육의 화학적 조성을 측정하였다. 혈액 내 GOT와 GPT 수치, 총 콜레스테롤, Ca 및 P을 측정하였으며, 경골의 중량, 파쇄 강도, 회분 함량, Ca및 P 함량을 측정하였다. 증체량은 대조구에 비하여 rPST-yeast culture를 $0.1\%$ 첨가한 실험구에서 증가하는 경향을 보였으며, 사료섭취 량 및 사료요구율은 처리구간 큰 차이가 없었다. 비장 및 복강 지방의 상대적 중량은 실험구간 유의한 차이가 관찰되지 않았으나, 간과 가슴근육의 상대적 중량은 처리간에 유의한 차이가 관찰되었는데,특히 T2처리구에서 가슴근육의 상대적 중량이 대조구에 비해 유의하게 증가하는 결과가 관찰되었다(p<0.05). 가슴 근육 내 조단백질 및 조지방 함량에 처리구간에 큰 차이는 없었으나, 수분 함량은 대조구에 비해 rPST-yeast culture와 SC yeast culture 첨가 급여구에서 유의하게 증가하였다(P<0.05). 혈청 내 GOT 수치는 변화가 없었으나, GPT수치는 rPST-yeast culture 첨가 급여 후에 유의하게 감소하는 결과가 얻어졌다(P<0.05). 혈중 총 콜레스테롤, Ca 및 P 농도는 처리간에 큰 차이를 보이지 않았다. 경골중량, 파쇄강도 및 화학적 성분 조성에서도 처리간에 큰 차이는 나타나지 않았다. 본 연구의 결과로부터 사료 내 rPST-yeast culture의 첨가급여에 의해 육계에서 성장성적이 향상될 가능성이 제시되었고, 특히 가슴근육이 증가와 같은 긍정적인 효과를 기대할 수 있을 것으로 사료되었다.

• Journal of Periodontal and Implant Science
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• 제50권1호
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• pp.56-66
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• 2020

Purpose: A stability-measuring device that utilizes damping capacity analysis (DCA) has recently been introduced in the field of dental implantology. This study aimed to evaluate the sensitivity and reliability of this device by measuring the implant stability of ex vivo samples in comparison with a resonance frequency analysis (RFA) device. Methods: Six implant beds were prepared in porcine ribs using 3 different drilling protocols to simulate various implant stability conditions. Thirty-six pork ribs and 216 bone-level implants measuring 10 mm in height were used. The implant beds were prepared using 1 of the following 3 drilling protocols: 10-mm drilling depth with a 3.5-mm-diameter twist drill, 5-mm drilling depth with a 4.0-mm-diameter twist drill, and 10-mm drilling depth with a 4.0-mm-diameter twist drill. The first 108 implants were external-connection implants 4.0 mm in diameter, while the other 108 implants were internal-connection implants 4.3 mm in diameter. The peak insertion torque (PIT) during implant placement, the stability values obtained with DCA and RFA devices after implant placement, and the peak removal torque (PRT) during implant removal were measured. Results: The intraclass correlation coefficients (ICCs) of the implant stability quotient (ISQ) results obtained using the RFA device at the medial, distal, ventral, and dorsal points were 0.997, 0.994, 0.994, and 0.998, respectively. The ICCs of the implant stability test (IST) results obtained using the DCA device at the corresponding locations were 0.972, 0.975, 0.974, and 0.976, respectively. Logarithmic relationships between PIT and IST, PIT and ISQ, PRT and IST, and PRT and ISQ were observed. The mean absolute difference between the ISQ and IST values on a Bland-Altman plot was -6.76 (-25.05 to 11.53, P<0.05). Conclusions: Within the limits of ex vivo studies, measurements made using the RFA and DCA devices were found to be correlated under a variety of stability conditions.