• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.36 no.2
• /
• pp.87-93
• /
• 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.

• Journal of Embryo Transfer
• /
• v.32 no.3
• /
• pp.209-220
• /
• 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
• /
• v.34 no.4
• /
• pp.546-557
• /
• 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.

• Journal of Life Science
• /
• v.17 no.10
• /
• pp.1315-1320
• /
• 2007

Epiphyseal development in the long bones was studied radiographically in minipigs. Radiographs of the proximal and distal epiphyses of humerus, radius, ulna, femur and tibia were obtained at 4, 8, 12, 20, 40, 48, 96 and 144 weeks of age in total 58 minipigs. The assessment of maturity process was made in accordance with the criteria proposed by Owada and Sutow. The secondary ossification centers developed rapidly from 4 weeks of age to 40 weeks of age, and gradually thereafter until 96 weeks of age. The earliest epiphyseal fusion was apparent in the proximal radius, proximal and distal femur at 96 weeks of age. The complete fusion of the epiphyseal line in the long bones was evident on 144 weeks of age and was observed in most long bones such as the proximal humerus, the proximal and distal ulna and the distal radius, and the proximal tibia in minipigs.

• Journal of Periodontal and Implant Science
• /
• v.51 no.3
• /
• pp.179-188
• /
• 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
• /
• v.30 no.3
• /
• pp.157-162
• /
• 2006

Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

• Journal of Korean Orthopaedic Sports Medicine
• /
• v.8 no.1
• /
• pp.26-32
• /
• 2009

Purpose: This study was performed to compare the strength of ligamentous attached sites of cadaveric distal femur and to obtain reliable biomechanical data to use in ligamentous reconstruction or augmentation. Materials and Methods: Fifteen cadaveric distal femurs were used for this study. After measuring the bone density, 5.0 mm cannulated screw (Experiment 1) or reconstructed porcine ligament (Experiment 2) was inserted into the each ligamentous attached sites of anterior cruciate ligament (ACL), posterior cruciate ligament (PCL), medial collateral ligament (MCL) and lateral collateral ligament (LCL). In experiment 2, reconstructed porcine graft was fixed with bioabsorbable screw in ligamentous insertion sites. And we measured the maximal pullout force of each ligamentous attached sites of cadaveric distal femur. Results: Average bone mineral density was $1.205{\pm}0.137\;g/cm^2$ in experiment 1, $1.236{\pm}0.089\;g/cm^2$ in experiment 2, which showed no statistically significant differences. In experiment 1, average pull-out strength of ACL, PCL, MCL and LCL group were $519.1{\pm}111.7$ N, $638.9{\pm}144.4$ N, $169.7{\pm}56.0$ N, $225.6{\pm}61.5$ N respectively. In experiment 2, the average pull-out strength were $310.6{\pm}31.0$ N, $379.9{\pm}47.4$ N, $104.0{\pm}14.4$ N, $131.5{\pm}21.9$ N respectively. In experiment 1, there was no significant difference between ACL and PCL group and between MCL and LCL group. However, the maximal pullout strength of MCL and LCL group were significantly lower than that of ACL and PCL group (p<0.01). Experiment 2 showed the same results of experiment 1. Conclusion: Because stiffness of MCL and LCL attached sites are much lower than that of ACL and PCL attached sites, we may consider augmented fixation in ligamentous reconstructions of MCL and LCL.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.7
• /
• pp.925-937
• /
• 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.

• Korean Journal of Poultry Science
• /
• v.31 no.4
• /
• pp.245-253
• /
• 2004

This study was conducted to investigate the effects of dietary yeast culture containing rPST on growth performances and the characteristics of body compositions in broiler chickens. A total of 460 Ross male broiler chicks aged 2-day-old were fed one of five experimental diets; Control(devoid of all), TI (with $0.1\%$ antibiotics; chlorotetracycline), T2 (with $0.1\%$ rPST-yeast culture), T3 (with $0.2\%$ rPST-yeast culture) or T4 (with $0.2\%$ SC yeast culture) for 6 weeks. Feed consumption and body weights were measured weekly. At week 5 of the experiment, 10 chicks were randomly selected and sacrificed. The relative weights of each organ and the chemical composition of edible meat were measured. The enzyme activity, total cholesterol, Ca and P were also determined. Tibial weight, bone strength and chemical composition were investigated. There were no significant differences in feed intake and feed conversion ratios among the treatments throughout the experimental period. Body weight gains tended to be increased by feeding of diets containing rPST-yeast culture. The relative weight of breast muscle in T3 group was significantly higher than that of the control (P<0.05). The moisture contents of breast meat in groups fed diets containing rPST-yeast culture or SC yeast culture were significantly increased as compared with those of the control and n. However, the contents of crude protein and ether extract were not affected by feeding of rPST-yeast culture. There were no significant differences in GOT, total cholesterol, Ca and P. The relative weight and strength and proximal composition of the tibia were also not affected by dietary treatments. These results indicated that dietary rPST-yeast culture may be a valuable alternative for optimizing growth performances, particularly for improving the yield of breast muscle.

• Journal of Periodontal and Implant Science
• /
• v.50 no.1
• /
• pp.56-66
• /
• 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.