• Journal of Chest Surgery
• /
• v.23 no.4
• /
• pp.622-639
• /
• 1990

The use of aortic valve homograft has been developed since 1962 when Ross and Barratt - Boyes independently replaced a diseased aortic valve with an orthotopically inserted homograft valve. And also surgical treatment of complex congenital cardiac malformations utilizing homograft extracardiac conduit has been tried with better result than any other prosthetic material. The present study was undertaken to clarify the safety tissue viability, sterility, after following our protocol of procurement of heart, dissection of aortic and pulmonic homograft, sterilization, cryopreservation, thawing and dilution, and transplantation on experimental animal, sheep. Tissue viability of valve and great artery was assessed by tissue culture. Sterility was evaluated by bacterial and fungal culture. The method used was proven no deleterious effect on the integrity of the valve. Tissue culture of valve tissue before, and after cryopreservation process resulted that active fibroblast growth was observed from homograft sterilized with antibiotics. And culture of the transplanted homograft from sacrificed animal showed active fibroblast growth. Pathologic examination of implanted valve tissue from sacrificed sheep showed mild calcification and minor change, but there were moderate and severe calcification of wall of great arteries.

• Biomaterials Research
• /
• v.22 no.4
• /
• pp.311-318
• /
• 2018

Background: Tissue regeneration includes delivering specific types of cells or cell products to injured tissues or organs for restoration of tissue and organ function. Stem cell therapy has drawn considerable attention since transplantation of stem cells can overcome the limitations of autologous transplantation of patient's tissues; however, it is not perfect for treating diseases. To overcome the hurdles associated with stem cell therapy, tissue engineering techniques have been developed. Development of stem cell technology in combination with tissue engineering has opened new ways of producing engineered tissue substitutes. Several studies have shown that this combination of tissue engineering and stem cell technologies enhances cell viability, differentiation, and therapeutic efficacy of transplanted stem cells. Main body: Stem cells that can be used for tissue regeneration include mesenchymal stem cells, embryonic stem cells, and induced pluripotent stem cells. Transplantation of stem cells alone into injured tissues exhibited low therapeutic efficacy due to poor viability and diminished regenerative activity of transplanted cells. In this review, we will discuss the progress of biomedical engineering, including scaffolds, biomaterials, and tissue engineering techniques to overcome the low therapeutic efficacy of stem cells and to treat human diseases. Conclusion: The combination of stem cell and tissue engineering techniques overcomes the limitations of stem cells in therapy of human diseases, and presents a new path toward regeneration of injured tissues.

• The Korean Journal of Physiology and Pharmacology
• /
• v.13 no.1
• /
• pp.15-22
• /
• 2009

This study was undertaken to elucidate the underlying mechanisms of ATP depletion-induced membrane transport dysfunction and cell death in renal proximal tubular cells. ATP depletion was induced by incubating cells with 2.5 mM potassium cyanide(KCN)/0.1 mM iodoacetic acid(IAA), and membrane transport function and cell viability were evaluated by measuring $Na^+$-dependent phosphate uptake and trypan blue exclusion, respectively. ATP depletion resulted in a decrease in $Na^+$-dependent phosphate uptake and cell viability in a time-dependent manner. ATP depletion inhibited $Na^+$-dependent phosphate uptake in cells, when treated with 2 mM ouabain, a $Na^+$ pump-specific inhibitor, suggesting that ATP depletion impairs membrane transport functional integrity. Alterations in $Na^+$-dependent phosphate uptake and cell viability induced by ATP depletion were prevented by the hydrogen peroxide scavenger such as catalase and the hydroxyl radical scavengers(dimethylthiourea and thiourea), and amino acids(glycine and alanine). ATP depletion caused arachidonic acid release and increased mRNA levels of cytosolic phospholipase $A_2(cPLA_2)$. The ATP depletion-dependent arachidonic acid release was inhibited by $cPLA_2$ specific inhibitor $AACOCF_3$. ATP depletion-induced alterations in $Na^+$-dependent phosphate uptake and cell viability were prevented by $AACOCF_3$. Inhibition of $Na^+$-dependent phosphate uptake by ATP depletion was prevented by antipain and leupetin, serine/cysteine protease inhibitors, whereas ATP depletion-induced cell death was not altered by these agents. These results indicate that ATP depletion-induced alterations in membrane transport function and cell viability are due to reactive oxygen species generation and $cPLA_2$ activation in renal proximal tubular cells. In addition, the present data suggest that serine/cysteine proteases play an important role in membrane transport dysfunction, but not cell death, induced by ATP depletion.

• Archives of Plastic Surgery
• /
• v.36 no.2
• /
• pp.127-134
• /
• 2009

Purpose: Adipose tissue injection as a free graft for the correction of soft - tissue deficiency or depression deformity is a widespread procedure in plastic surgery. This study is to analyze the changes and viability of cryopreserved adipose tissue and to find out efficient long - term storage period. Methods: After centrifugation of aspirated abdominal tissues, $10m{\ell}$ of packed Adipose tissue were freezed at $-20^{\circ}C$. For 2, 4, 6, 8 months, each frozen samples were taken and injected into scalp of SCID mice. After 15 weeks, injected Adipose tissue were sampled and analyzed at 2 months interval. We compared and analyzed each group about the weight of the injected fat, histologic impressions, activity of mitochondria, size of a fat cell and rate of survival. Results: Significant weight changes were observed in cryopreservation for 2 months(p<0.05). Histologic changes were observed, independent of the freezing period with H - E stain. Among cryopreservations for 2, 4, 6 months, no significant change were observed. The reduction of mitochondrial enzymatic activity was observed independent of time interval but activity of mitochondrial dehydrogenase was reduced less than 50% in MTT assay. Conclusion: Freezing in $-20^{\circ}C$ for 6 months has no adverse effect to Adipose tissue, but fragile adipocytes, damaged cell membrane during harvesting procedure, were disrupted within 1 - 2 month and the maximum volume reduction were followed less than 2 months. These results demonstrate that tissue preparation cells without membrane damage have the greatest viability level and cryopreservation less than 2 months has great volume effect and cryopreservation for 6 months has stable volume effect.

• Journal of Korean Dental Science
• /
• v.14 no.2
• /
• pp.51-60
• /
• 2021

Purpose: Measuring viability of a three-dimensional in vitro organotypic human oral tissue model has been suggested as an alternative test method to the oral mucosa irritation test of oral care products. The aim of this study was to investigate the production of two different cytokines using organotypic human oral tissue model following exposure to chemicals that are commonly used in oral care products. Materials and Methods: The organotypic human oral tissues were exposed to ethanol, sodium lauryl sulphate or hydrogen peroxide for 90 minutes. Following exposure, interleukin (IL)-1α and IL-8 productions were assessed and correlated with cell viability testing as well as histology of the organotypic human oral tissues. Result: High levels of IL-8 were released from organotypic human oral tissues in all of the test and control groups without any significant differences between them. In contrast, differences were found in IL-1α release between the test and control groups. Additionally, the trend of IL-1α release corresponded to the phenotypes observed in histological analysis while different trend existed between IL-1α release and cell viability. Conclusion: The study concluded the non-specific release of IL-8 for the assessment of oral care product chemicals' toxicity, while potential of measuring IL-1α cytokine level as the possible alternative test method.

• The Journal of Korean Physical Therapy
• /
• v.21 no.3
• /
• pp.87-93
• /
• 2009

Purpose: TThis study examined the effect of repetitive magnetic stimulation (RMS) on the viability and proliferative response of human adipose tissue-derived stromal cells (hATSCs) in vitro. Methods: The hATSCs were cultured primarily from human adipose tissue harvested by liposuction and incubated in a $37^{\circ}C$ plastic chamber. The cells were exposed to a repetitive magnetic field using a customized magnetic stimulator (Biocon-5000, Mcube Technology). The RMS parameters were set as follows: repetition rate=10Hz, 25Hz (stimulus intensity 100%= 0.1 Tesla, at 4cm from the coil), stimulated time= 1, 5, and 20 minutes. Twenty four hours after one application of RMS, the hATSCs were compared with the sham stimulation, which were kept under the same conditions without the application of RMS. The cells were observed by optical microscopy to determine the morphology and assessed by trypan blue staining for cell proliferation. The apoptosis and viability of the hATSCs were also analyzed by fluorescence-activated cell sorting (FACS) analysis of Annexin V and MTT assay. Results: After RMS, the morphology of the hATSCs was not changed and the apoptosis of hATSCs were not increased compared to the sham stimulation. The viability of the cells was similar to the cells given the sham stimulation. Interestingly, the level of hATSC proliferation was significantly higher in all RMS groups. Conclusion: The application of RMS may not cause a change in morphology and viability of hATSCs but can increase the level of cell proliferation in vitro. RMS might be useful as an adjuvant tool in combination with stem cell therapy without adverse effects.

• Archives of Plastic Surgery
• /
• v.41 no.2
• /
• pp.126-132
• /
• 2014

Background Extensive degloving injuries of the extremities usually result in necrosis of the flap, necessitating comprehensive skin grafting. Provided there is a sufficient tool to evaluate flap viability, full-thickness skin can be used from a nonviable avulsed flap. We used a Wood's lamp to determine the viability of avulsed flaps in the operation field after intravenous injection of fluorescein dye. Methods We experienced 13 cases during 16 months. Fifteen minutes after the intravenous injection of fluorescein dye, the avulsed skin flaps were examined and non-fluorescent areas were marked under Wood's lamp illumination. The marked area was defatted for full-thickness skin grafting. The fluorescent areas were sutured directly without tension. The non-fluorescent areas were covered by defatted skin. Several days later, there was soft tissue necrosis within the flap area. We measured necrotic area and revised the flap. Results Among all the cases, necrotic area was 21.3% of the total avulsed area. However, if we exclude three cases, one of a carelessly managed patient and two cases of the flaps were inappropriately applied, good results were obtained, with a necrotic area of only 8.4%. Eight patients needed split-thickness skin grafts, and heel pad reconstruction was performed with free flap. Conclusions A full-thickness skin graft from an avulsed flap is a good method for addressing aesthetic concerns without producing donor site morbidity. Fluorescein dye is a useful, simple, and cost-effective tool for evaluating flap viability. Avulsed flap injuries can be managed well with Wood's lamp illumination and a full-thickness skin graft.

• The Journal of Advanced Prosthodontics
• /
• v.9 no.6
• /
• pp.453-462
• /
• 2017

PURPOSE. The purpose of this study was to investigate the in vitro cytotoxicity of thermoplastic denture base resins and to identify the possible adverse effects of these resins on oral keratinocytes in response to hot water/ food intake. MATERIALS AND METHODS. Six dental thermoplastic resin materials were evaluated: three polyamide materials (Smile tone, ST; Valplast, VP; and Luciton FRS, LF), two acrylic materials (Acrytone, AT; and Acryshot, AS), and one polypropylene resin material (Unigum, UG). One heat-polymerized acrylic resin (Vertex RS, RS) was chosen for comparison. After obtaining extracts from specimens of the denture resin materials (${\phi}=10$ mm and d=2 mm) under different extraction conditions ($37^{\circ}C$ for 24 hours, $70^{\circ}C$ for 24 hours, and $121^{\circ}C$ for 1 hour), the extracts (50%) or serial dilutions (25%, 12.5%, and 6.25%) in distilled water were co-cultured for 24 hours with immortalized human oral keratinocytes (IHOKs) or mouse fibroblasts (L929s) for the cytotoxicity assay described in ISO 10993. RESULTS. Greater than 70% viability was detected under all test conditions. Significantly lower IHOK and L929 viability was detected in the 50% extract from the VP ($70^{\circ}C$) and AT ($121^{\circ}C$) samples (P<.05), but only L929 showed reduced viability in the 50% and 25% extract from LF ($37^{\circ}C$) (P<.05). CONCLUSION. Extracts obtained from six materials under different extraction conditions ($37^{\circ}C$, $70^{\circ}C$, and $121^{\circ}C$) did not exhibit severe cytotoxicity (less than 70% viability), although their potential risk to oral mucosa at high temperatures should not be ignored.

• Clinical and Experimental Reproductive Medicine
• /
• v.26 no.2
• /
• pp.251-256
• /
• 1999

The present study was conducted to evaluate whether vitrification could be used for ovarian tissue preservation. The important issue here is that the vitrification is very simple, easy, and economical compared to the conventional cryopreserving method that using automatic freezing instrument. Human ovarian cortical tissues were cryopreserved by vitrification with 5.5 M ethylene glycol and 1.0 M sucrose as cryoprotectant. Three points of temperature ($4^{\circ}C$, room temperature, and $37^{\circ}C$) and two points of duration (5 or 10 minutes) for cryoprotectant treatment were examined to determine the best condition for vitrification of the human ovarian cortical tissues. After thawing, viability of the isolated primordial follicles was examined by dye-exclusion method. Histological appearance of tissues before and after the cryopreservation was evaluated. There was no toxic effect of the 5.5 M ethylene glycol on the primordial follicles. However, when the tissues were treated with cryoprotectant at $37^{\circ}C$ for 10 minutes and exposed to liquid nitrogen, it seems likely that there is certain deleterious effects on the viability of the primordial follicles. The highest viability of the primordial follicles was obtained with the treatment of cryoprotectant at room temperature for 10 minutes. Follicles and oocytes survived after freezing and thawing had the similar normal shapes as was seen in the specimens before cryopreservation. It would be useful to apply vitrification in establishing ovarian tissue banking for clinical purposes.

• Proceedings of the Korean Society of Applied Pharmacology
• /
• 1996.04a
• /
• pp.225-227
• /
• 1996