• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.97-102
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• 2013

The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

• Journal of Chest Surgery
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• v.36 no.4
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• pp.219-228
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• 2003

Background:Liquid nitrogen freezing techniques have already met with widespread success in biology and medicine as a means of long-term storage for cells and tissues. The use of cryoprotectants such as glycerol and dimethylsulphoxide to prevent ice crystal formation, with carefully controlled rates of freezing and thawing, allows both structure and viability to be retained almost indefinitely. Cryopreservation of various tissues has various con-trolled rates of freezing. Material and Method: To find the optimal freezing curve and the chamber temperature, we approached the thermodynamic calculation of tissues in two ways. One is the direct calculation method. We should know the thermophysical characteristics of all components, latent heat of fusion, area, density and volume, etc. This kind of calculation is so sophisticated and some variables may not be determined. The other is the indirect calculation method. We performed the tissue freezing with already used freezing curve and we observed the actual freezing curve of that tissue. And we modified the freezing curve with several steps of calculation, polynomial regression analysis, time constant calculation, thermal response calculation and inverse calculation of chamber temperature. Result: We applied that freezing program on mesenchymal stem cell, chondrocyte, and osteoblast. The tissue temperature decreased according to the ideal freezing curve without temperature rising. We did not find any differences in survival. The reason is postulated to be that freezing material is too small and contains cellular components. We expect the significant difference in cellular viability if the freezing curve is applied on a large scale of tissues. Conclusion: This program would be helpful in finding the chamber temperature for the ideal freezing curie easily.

• Polymer(Korea)
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• v.32 no.5
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• pp.403-408
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• 2008

Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

• Archives of Reconstructive Microsurgery
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• v.8 no.1
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• pp.28-34
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• 1999

The need for reconstruction of large bone, soft tissue defect of mandible has greater emphasis due to development of industry, traumatic accident and increase of tumor. The mandibular reconstruction had greatly progressed through the first and the second World Wars. The Fibular free flap by using microscope was reported in 1970 and many maxillofacial reconstructive surgeons had used. In 1988, Dr. Hidalgo first reported mandibular reconstruction by using fibular free flap. Mandibular reconstruction by using fibular free flap has several advantages. First, it provides up to 25 cm of bone, enough to reconstruct any length of mandible defect. Second, a skin island, based on a septocutaneous blood supply, is available in a size large enough to simultaneously reconstruct internal and external soft tissue defect. Third, The fibular donor site morbidity is low, fourth, it provides a esthetic effect of mandible line. And finally bone viability is good. The Fibular osteocutaneous free flap was performed after COMMANDO operation due to squamous cell cancer in oral cavity (15 cases). Therefore we report out successful operation of the mandible reconstruction by using fibular osteocutaneous free flap.

• Archives of Plastic Surgery
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• v.32 no.3
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• pp.319-326
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• 2005

This study was designed to investigate the optimal period of pedicles implantation in the prefabricated periosteofascial flap using a vascular tissue transfer. Flap prefabrication was prepared with a transposition of the central pedicles of right auricle on the calvarium of the New Zealand white rabbit. Thirty flaps were divided into five groups of six flaps, including control group (group I) of the conventional periosteofascial flap based on the right lateral border of parietal bone. The prefabricated flap was elevated as a $2{\times}2cm$ sized island flap and reposed in place in 1, 2, 3, and 4 weeks after the pedicles transfer in groups II, III, IV, and V, respectively. Five days after flap repositioning, the flap viability and vascularity were evaluated with microangiography and histological study quantitatively. The flap survival was increased in accordance with the implanted period of the pedicle. New vessels developed around the implanted pedicle in the 2nd week, and overall vascularization of the flap was accomplished in the 3rd week. The flap with 4 weeks of implantation period, however, showed the same survival rate as the control group. In conclusion, prefabricated periosteo- fascial flap can be created with a vascular tissue transfer, and the optimal duration of the pedicle implantation is more than 4 weeks to obtain adequate flap survival.

• KSBB Journal
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• v.19 no.3
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• pp.174-177
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• 2004

A method of collection and long-term storage of viable lily (Lilium longiflorum) pollen grains were developed for their in vitro growth and transformation in consistency. Petroleum ether, n-heptane, cyclohexane and benzene, as pollen collection medium, were determined less toxic to pollen growth in vitro than others tested. Pollen grains, however, lost their growth activity if stored in these solvents more than a week, So, a serial performance, that is, pollen grain collection in these solvents, air-drying and immediate transfer to low temperature condition was determined desirable for keeping the viability much longer. Pollen grains from this storage showed a successful transformation in vitro with a cDNA encoding tissue plasminogen activator (TPA) protein using Agrobacterium via vacuum infiltration according to western blotting analysis.

• Archives of Plastic Surgery
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• v.47 no.4
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• pp.340-346
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• 2020

Background Adipofascial flaps covered with a skin graft address the challenges involved in reconstructing dorsal foot defects. The purpose of this study was to describe a large adipofascial flap based on the perforators of the dorsalis pedis artery for large foot defects. Methods Twelve patients aged 5-18 years with large soft tissue defects of the dorsal foot due to trauma were treated with an extended dorsalis pedis adipofascial flap from May 2016 to December 2018. The flap was elevated from the non-injured half of the dorsum of the foot. Its length was increased by fascial extension from the medial or lateral foot fascia to the plantar fascia to cover the defect. All perforators of the dorsalis pedis artery were preserved to increase flap viability. The dorsalis pedis artery and its branches were kept intact. Results The right foot was affected in 10 patients, and the left foot in two patients. All flaps survived, providing an adequate contour and durable coverage with a thin flap. Follow-up lasted up to 2 years, and patients were satisfied with the results. They were able to wear shoes. Donor-site morbidity was negligible. Two cases each of partial skin graft loss and superficial necrosis at the tip of the donor cutaneous flap occurred and were healed by a dressing. Conclusions The hinged multiperforator-based extended dorsalis pedis adipofascial flap described herein is a suitable method for reconstructing dorsal foot defects, as it provides optimal functional and aesthetic outcomes with minimal donor site morbidity.

• Archives of Reconstructive Microsurgery
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• v.3 no.1
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• pp.40-44
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• 1994

Microsurgery has advanced beyond its nascent stages reaching success rates of 90% to 95%. However, this means that even in the best circumstances, 5% to 10% of free flaps and replants fail. Almost all failures are due to vessel thrombosis, resulting in ischemia of the transplanted tissue. Many attemps have been undertaken to treat and reverse its effects. Zdeblick and colleagues noted an improvement in the viability of amputated limbs replanted after an extended period of ischemia following intraarterial infusion of urokinase. Subsequent studies have investigated many modalities of urokinase administration in various animal models by differing ischemic periods. These studies, however, have failed to establish a definitive, generally accepted protocol for administration of urokinase in the salvage of tissue subjected to prolonged ischemia. Our clinical observations suggest that a bolus of urokinase delivered under pressure may increase the thromoblytic effect of the drug, probably by means of increased delivery to microvasculature. We intend to investigate the role of selective pressure perfusion of ischemic flaps as a new means for increasing the effectiveness of urokinase in the treatment of the "no-reflow" phenomenon. A total of 32 male New Zealand rabbits were used and divided into the four groups according to the method of infusion. After 12 hours of ischemia the flaps were injected with Hartmann's solution or with urokinase and the percent survival of the flap was determined at 7 days following flap reperfusion. As the result, the flap survival rate was highest in the pressure injection of urokinase group.

• Archives of Pharmacal Research
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• v.26 no.10
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• pp.855-860
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• 2003

The effect of silver sulfadiazine (SSD) on the proliferation of human dermal fibroblast (HDF) was studied to determine the impact of the drug on the wound healing process and dermal mechanical strength. Human dermal fibroblasts were cultured to 80% confluency using DMEM with 10% FBS and viability of the cell was estimated using neutral red assay. In addition, the $2^{nd}$ degree burn wound was prepared on the anterior part of rabbit ear skin and dressings containing SSD were applied for 96 h. Presence of inflammatory cells and degree of re-epithelialization were investigated in the wound. After 15 day of the induction of burn wounds, the treated area was excised and dermal mechanical strength was quantitatively measured with a constant speed tensiometer. SSD was found to be highly cyto-toxic in cultured HDF cells. The topical application of SSD (2%) could control the infection as evidenced by the lack of accumulation of inflammatory cells in histological evaluation. Therefore, these observations suggested that the impairment of dermal regeneration and decreased mechanical strength of dermal tissue was resulted from the cyto-toxic effect of SSD on dermal cells. Since the decreased mechanical strength may lead to reduction in resilience, toughness and maximum extension of the tissue, the identification of optimum dose for SSD that limits infection while minimizes the cyto-toxic effect may be clinically relevant.

• International Journal of Oral Biology
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• v.46 no.1
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• pp.51-59
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• 2021

Periodontal disease is an inflammatory disease that affects the destruction of the bone supporting the tooth and connective tissues surrounding it. Periodontal ligament fibroblasts (PDLFs) induce overexpression of matrix metalloproteinase (MMP) involved in periodontal disease's inflammatory destruction. Osteoclasts take part in physiological bone remodeling, but they are also involved in bone destruction in many kinds of bone diseases, including osteoporosis and periodontal disease. This study examined the effect of baicalin on proteolytic enzymes' production and secretion of inflammatory cytokines in PDLFs and RAW 264.7 cells under the lipopolysaccharide (LPS)-induced inflammatory conditions. Baicalin inhibited the expression of the protein, MMP-1 and MMP-2, without affecting PDLFs' cell viability, suggesting its possibility because of the inhibition of phosphorylation activation of mitogen-activated protein kinase's p38, and the signal transduction process of nuclear factor κB (NFκB)-related protein. Also, baicalin reduced the expression of MMP-8 and MMP-9 in RAW 264.7 cells. This reduction is thought to be due to the inhibition of the signal transduction process of NFκB-related proteins affected by inhibiting p65RelA phosphorylation. Also, baicalin inhibited the secretion of nitric oxide and interleukin-6 induced by LPS in RAW 264.7 cells. These results suggest that baicalin inhibits connective tissue destruction in periodontal disease. The inhibition of periodontal tissue destruction may be a therapeutic strategy for treating inflammatory periodontal-diseased patients.