• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.21-27
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• 2004

This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS ＋ 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P＜0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P＜0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P＜0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

• Archives of Plastic Surgery
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• v.34 no.6
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• pp.685-690
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• 2007

Purpose: Numerous materials, both autologous and nonautologous, have been used for augmentation of sunken areas and each has its own limitations. The ideal material for augmentation should not be absorbed in any manner. This study is designed to assess the survival of $SureDerm^{(R)}$, $Permacol^{(R)}$ graft according to the volume and histologic change. Methods: Twenty four mice, weighing about 50 grams and of 5 weeks of age were used. $SureDerm^{(R)}$ is an acellular dermal matix obtained from human cadeveric skin. $Permacol^{(R)}$ is a porcine derived acellular dermal matrix whose manufacture involves trypsinisation, solvent extraction. Graft pieces standardized to $1{\times}1cm$ size were used in each group. The implanted material were taken 1, 4, 8 and 12 weeks later, respectively. The changes of graft volume during the graft period were measured on initial, 1, 4, 8 and 12 weeks. Results: The initial shape of graft was maintained up to 12 weeks in $Permacol^{(R)}$ graft group and mean survival rate was $80.36{\pm}8.21%$ in $SureDerm^{(R)}$, $89.57{\pm}6.39%$ in $Permacol^{(R)}$(p=0.01). The volume of each graft decreased 29% from initial volume on 12 weeks in $SureDerm^{(R)}$, 18% in $Permacol^{(R)}$. The structure of $Permacol^{(R)}$ remained until 12 week after implantation. Conclusion: Our experimental study suggests that $Permacol^{(R)}$ could be a safe material as an implant for permanent augmentation. However, There are further study remained for antigenicity of these material, and the choice of graft for augmentation should be remained to the clinical situations.

• Journal of Korean Medicine for Obesity Research
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• v.11 no.1
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• pp.47-60
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• 2011

Objectives The objective of this study was to contribute to the development of pharmacupuncture for obesity treatment by reviewing the studies of pharmacupuncture experimentations and clinical trials. Methods We searched the papers with keywords of 'obesity' and 'pharmacupuncture' in the search site, RISS, Oriental medicine advanced searching integrated system(OASIS), Korean traditional knowledge portal, the society of Korean medicine for obesity research, Korean pharmacupuncture institute, the Korean academy of oriental rehabitation medicine and Korean acupuncture & moxibustion society. Results 1. We reviewed 37 articles searched. 27 articles(73.0%) were animal experimentations, 5 articles(13.5%) were cell experimentations, 4 articles(10.8%) were clinical trials and 1 article(2.7%) was study analysis. 2. The herbs, using for animal experimentations, were atratylodes japonica, coix lachrymajobi, ephedra sinica, crataegus pinnatifida, wild ginseng and etc. Acupucture points were joksamni(ST36), zhongwan(CV12), gansoo(BL18), pungnyung(ST40), umnungchon(SP9), bisu(BL20), gokji (LI11), cheun-chu(ST25) and etc. 3. For cell experimentations, preadipocytes and adipocytes performed on cell cultures with using rats, 3T3-L1 preadipocytes and porcine skin including fat tissue were treated with fel ursi, bovis calculus, ephedrae herba, spirodelae herba, wild ginseng. 4. For clinical trials, Sangsik no.1, Bigiheo, ephedra, green tea and sweet bee venom were injected at the region where a lot of fat like zhongwan(CV12), xiawan(CV10), kwanwon(CV4), cheun-chu(ST25) and thigh. Conclusion Through animal and cell experimentations and clinical trials, the treatment of obesity using local acupuncture therapy was effective. For clinical use, however, it is considered that animal and cell experimentation and clinical trial's connection using one kind of herb and studies about more clinical trials and associated side effects are needed.

• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.267-271
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• 1997

We developed a dermal equivalent (DE) which was engineered using human dermal fibroblasts and a matrix of collagen gel. The in vitro construction of the DE was accomplished by casting a porcine collagen type I solution plus concentrated medium with isolated and cultured fibroblasts. These constructs were attached to culture dishes or left floating in culture medium. Contraction of attached gels results in decreased gel thickness without a change in gel diameter, and contraction of floating gels results in decreased gel thickness and diameter. After contraction, there was no increase in cell number in floating gels, but cells in attached gels began to increase after about 4 days of the lag phase in cell growth curve. At this lag phase, addition of fibroblast growth factor (FGF) at a concentration of $0.1{\mu}$/ml promoted cell proliferation in the attached collagen gels, but no effect in floating gels. These results indicate that the method of contraction had an influence on the extracellular matrix (ECM) organization, and this influenced not only cell growth but also fibroblast responsiveness to FGF. This suggests that attached collagen gel is more suitable as a dermal equivalent than the floating gel. And the final contracted area of attached gel is much larger than that of the floating gel since floating gel is contracted in all directions but attached gel is contracted only vertically.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.1
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• pp.69-88
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• 1999

Several solvent extracts from Alpinia katsumudai were prepared and their various anti-inflammatory activities were evaluated. Alpinia katsumadai extract showed high various anti-inflammatory effects among the 8 medicinal plant extracts, Butanol extract from Alpinia katsumndai showed a potent anti-oxidative and free radical scavenging activities, Free radical scavenging effect of butanol fraction of Alpinia katsumadai($IC_{50}$/ : 50$\mu$g/m$\ell$) was higher than butylated hydroxytoluene($IC_{50}$/ : 50$\mu$g/m$\ell$) and ascorbic acid($IC_{50}$/ : 22$\mu$g/m$\ell$). Alpinia katsumadai butanol fraction exhibited relatively high antioxidative activity($IC_{50}$/ : 335$\mu$g/m$\ell$) compared to ascorbic acid. The inhibitory effect of Alpinia katsumadal ethanol extract on elastase exhibited 10 to 78% at 100 to 1000$\mu$g/m$\ell$ concentration , tile $IC_{50}$/ values with 465.7$\mu$g/m$\ell$ for porcine pancreatic elastase(PPE) and 481.9$\mu$g/m$\ell$ for human leukocyte elastase(HLE), respectively The Alpinta katsumadai extract inhibited effectively hyaluronidase activity($IC_{50}$/ 335$\mu$g/m$\ell$), and showed inhibition in vitro on delayed hypersensitivity when it was topically applied. These results suggest that Alpinia katsumadai extract may reduce inflammatory skin trouble. The Alpinia katsumadai extract also showed higher Inhibitory effect of melanin biosynthesis on cultured melanoma cell compared to arbutin and kojic acid.

• Polymer(Korea)
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• v.38 no.3
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• pp.391-396
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• 2014

Polymer microneedles can be fabricated by a micromolding method, an easy and cost-effective method. However, it is not easy to achieve uniform coating with an aqueous coating solution due to hydrophobic surface of polymer microneedles. 3-Dimensional coating polymer microneedles could deliver more than twice as much dose as in-plane metal microneedles by increasing coating area and the number of microneedles per unit area. A uniform coating was not obtained by addition of coating additives in the coating solution. The satisfied coating was achieved by treatment of surface of polymer microneedle with metal deposition and UV/ozone, and UV/ozone treatment was an ultimate surface treatment method based on biological safety. Calcein coating polymer microneedles were prepared by using UV/ozone treatment and followed dip-coating, and they delivered calcein in porcine skin successfully after 15 min of insertion.

• Journal of the Optical Society of Korea
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• v.17 no.4
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• pp.289-295
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• 2013

Although low-level laser therapy (LLLT) has been a valuable therapeutic technology in the clinic, its efficacy may be reduced in deep tissue layers due to strong light scattering which limits the photon density. In order to enhance the photon density in deep tissue layers, this study developed an optical tissue clearing (OTC) laser probe (OTCLP) system which can utilize four different OTC methods: 1) tissue temperature control from 40 to $10^{\circ}C$; 2) laser pulse frequency from 5 to 30 Hz; 3) glycerol injection at a local region; and 4) a combination of the aforementioned three methods. The efficacy of the OTC methods was evaluated and compared by investigating laser beam profiles in ex-vivo porcine skin samples. Results demonstrated that total (peak) intensity at full width at half maximum of laser beam profile when compared to control data was increased: 1) 1.21(1.39)-fold at $10^{\circ}C$; 2) 1.22 (1.49)-fold at a laser pulse frequency of 5 Hz; 3) 1.64 (2.41)-fold with 95% glycerol injection; 4) 1.86 (3.4)-fold with the combination method. In conclusion, the OTCLP system successfully improved the laser photon density in deep tissue layers and may be utilized as a useful tool in LLLT by increasing laser photon density.

• Archives of Plastic Surgery
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• v.38 no.5
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• pp.576-584
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• 2011

Purpose: The shape and location, the amount of the wound and the characteristics of the remaining tissues are known to influence wound contraction. The previous studies using small animals have not been an appropriate model because the wound healing mechanisms and skin structures are different from those of the human. The purpose of this study is to evaluate wound contraction according to the shape and location of the wound using a $Micropig^{(R)}$. Methods: Four $Micropigs^{(R)}$ (Medikinetics, Pyeongtaek, Korea) that were 10 months of age weighed 25 kg were used. Full thickness skin defects were made by clearing all the tissues above the fascial layer in the shape of square, a regular triangle and a circle of 9 $cm^2$ each on the back around the spine. Eight wounds were created on the back of each pig, 50 mm apart from each other. The randomly chosen wound shapes included 11 squares, 11 regular triangles, and 10 circles. Wound dressing was done every other day with polyurethane foam. The wound size was measured using a Visitrak $Digital^{(R)}$ (Smith & Nephew, Hull, UK) on every other day after surgery from day 2 to day 28. A biopsy was performed on day 3, and 1, 2, 3 and 4 weeks to investigate the degree of acute and chronic inflammation, the number of microvesssel and myofibroblast density using H & E stain and immunohistochemistry. The wound contraction rate was calculated to figure out the differences among each of the shapes and the locations. Results: The ultimate shape of the circle wound was oval, and that of the regular triangle and square were stellate. The maximum contraction rate was obtained on 8 to 10 days for all the shapes, which corresponds with the immunohistochemical finding that myofibroblast increases in the earlier 2 weeks whereas it decreases in the later 2 weeks. Epithelialization was seen in the wound margin on day 7 and afterwards. The final wound contraction rates were highest for the regular triangle shapes; however, there were no statistically significant differences. The wound contraction rates by locations showed statistically significant differences. The wound in the cephalic area presented more contractions than that of the wounds in the caudal area. Conclusion: The location of a wound is more important factor than the wound shape in wound contraction.

• Journal of Veterinary Clinics
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• v.27 no.6
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• pp.647-651
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• 2010

The advantages of the $CO_2$ laser are offset by the delay in laser wound healing secondary to thermal damage. To minimize the undesirable thermal damage of the $CO_2$ laser, investigators have developed technical advances in the delivery system of the laser energy. This study compared tissue healing of the continuous and the pulsed modes $CO_2$ laser wounds in an animal surgery model. Five healthy Landrace and Yorkshire mixed breeds of both genders were used (45-51 kg, 4-6 months old, three males and two females). A full thickness wound of skin ($2{\times}2{\times}2cm^2$) was made over on the each pig's both sides of dorsal midline at 0, 7, 14, and 18 days. The wounds created at 18, 14, 7 and 0 days were named post-wounding day (PWD) 3, 7, 14 and 21, respectively. In each pig, one wound (left side) was treated pulsed $CO_2$ laser and the other wound (right side) was treated continuous wave $CO_2$ laser. Each wound was closed with two interrupted suture of 3-0 sutures. At 21 days after initial wounding, each wound was taken for histological evaluations. The degrees of reepithelialization were performed more prominently in the pulsed mode group than in the continuous mode group. The degrees of granulation were greater significantly in pulsed mode group than those in the continuous mode on PWD 3 (p < 0.05). The degrees of fibroblasts in the pulsed mode group were greater significantly in comparison to those in the continuous mode group on PWD 7, (p < 0.05). In conclusion, reepithelialization, granulation and fibroblasts in the pulsed mode group were greater markedly in comparison to those in the continuous mode group. It was considered that pulsed mode $CO_2$ laser was more suitable for the skin incision than the continuous mode $CO_2$ laser.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.168-176
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• 2010

Acellular dermal matrix (ADM), produced by decellularization from human cadaveric skin, has been used for various biomedical applications. A manufacturing process for ADM ($SureDerm^{TM}$) using tri-n-butyl phospahate (TnBP) and deoxycholic acids as the decellularization solution has been developed. The manufacturing process for $SureDerm^{TM}$ has 70% ethanol treatment and ethylene oxide gas sterilization for inactivating infectious microorganisms. The purpose of this study was to examine the efficacy of the 70% ethanol treatment, decellularization process using 0.1% TnBP and 2% deoxycholic acids, and EO gas sterilization process in the inactivation of viruses. A variety of experimental model viruses for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), and porcine parvovirus (PPV) were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment. However HAV and PPV showed high resistance to 70% ethanol treatment with the log reduction factors of 1.85 and 1.15, respectively. HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by decellularization process. All the viruses tested were completely inactivated to undetectable levels by EO gas treatment. The cumulative log reduction factors of HIV-1, BHV, BVDV, HAV, and PPV were $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, and $\geq7.18$, respectively. These results indicate that the production process for $SureDerm^{TM}$ has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.