Lee, Sang-Woon;Park, Yong-Tae;Kim, Seong-Gon;Kweon, HaeYong;Jo, You-Young;Lee, Heui Sam
Maxillofacial Plastic and Reconstructive Surgery
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v.34
no.5
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pp.293-298
/
2012
Purpose: The aim of this study was to evaluate the bone regeneration ability of 1% tetracycline (TC)-loaded silk fibroin membrane (SFM), in a rabbit calvarial defect model. Methods: Twenty New Zealand white rabbits were used for this study. Bilateral round defects were made on the rabbit parietal bone, using trephine bur with an 8 mm diameter. TC-loaded SFM or SFM was covered on the right parietal bone defect, and the left parietal bone defects were uncovered for the control. The animals were humanely sacrificed at 4 or 8 weeks postoperatively. A micro-computerized tomography (${\mu}$-CT) of each specimen was taken for analysis of bone regeneration. Hematoxylin and Eosin stain were done to observe histological findings. Results: From the ${\mu}$-CT results, regenerated bone volume ($mm^3$) of 1% TC-loaded SFM, SFM, and control were $7.80{\pm}5.87$, $8.79{\pm}3.44$, and $10.61{\pm}5.3$ at 4 weeks postoperatively, respectively (P>0.05). Regenerated bone volume ($mm^3$) of 1% TC-loaded SFM, SFM, and control were $36.56{\pm}8.50$, $25.86{\pm}8.17$, and $19.09{\pm}5.07$ at 8 weeks postoperatively, respectively (P<0.05). Conclusion: The 1% TC-loaded SFM showed more bone regeneration than the SFM and the uncovered control, in guided bone regeneration.
In this study, we evaluated the potential of 70% ethanol extract from Persicaria nepalensis (PNE) as a cosmetic ingredient by primary skin irritation, ocular irritation, and maximization tests for delayed hypersensitivity in New Zealand white rabbits and Hartley guinea pig. Skin safety study was performed to evaluate the potential toxicity of PNE using the primary irritation test. In the primary irritation test, 50% PNE was applied to the skin, and no adverse reactions such as erythema and edema were observed at the intact skin sites. Therefore, PNE was classified as a practically non-irritating material based on a primary irritation index of "0.0.". In the ocular irritation test, the 50% PNE applied did not show any adverse reactions in the different parts of rabbit eyes, including the cornea, iris, and conjunctiva. Thus, PNE was classified as a practically non-irritating material based on an acute ocular irritation index of "0.0.". Skin sensitization was tested by the Guinea Pig Maximization Test (GPMT) and Freund's Complete Adjuvant (FCA) using an intradermal injection of 10% PNE. Edema and erythema were not observed 24 and 48 h after the topical application of PNE in skin sensitization test, which exhibited a sensitization score of "0.0.". Therefore, it can be suggested that P. nepalensis could be used as potential candidates for cosmoceutical ingredients, without any major side effects.
Purpose : This experiment examined the effects of anodization on commercially pure titanium implant fixtures. Material & methods : The implant fixtures were anodized at three different voltage levels, producing three different levels of oxidation on the surface of the fixure. Implant were divided into four groups according to the level of oxidation. Group 1 consist of the control group of machined surface implants, Group 2 implants were treated by anodizing to 100 voltage, Group 3 implants were treated by anodizing oxidation to 200 voltage Group 4 implants were treated by anodizing oxidation to 350 voltage. Surface morphology was observed by Scanning Electron Microscope(SEM) and the surface roughness was measured using NanoScan $E-1000^{\circledR}$. Implantation of the fixtures were performed using New Zealand white rabbits. $Periotest^{\circledR}$ value(PTV) resonance frequency analysis(RFA), and removal torque were measured in 0, 2, 4, 8, 12 weeks after implantation. Results : The results of the study were as follows: 1. Values for the measured surface roughness indicate statistically significant differences in Ra, Rq, and Rt values among group 1, 2, 3, and 4 at the top portion of the thread,(p<0.05) while values at the base of the threads indicated no significant difference in these values. 2. A direct correlation between the firming voltage, and surface roughness and irregularities were observed using scanning electron microscope. 3. No statistically significant differences were found between test groups regarding $Periotest^{\circledR}$ values. 4. Analysis of the data produced by RFA, significant differences were found between group 1 and group 4 at 12 weeks after implantation.(p<0.05) Conclusions : In conclusion, no significant differences could be found among test groups up to a certain level of forming voltage threshold, beyond this firming voltage threshold, statistically significant differences occurred as the surface area of the oxide layer increased with the increase in surface porosity, resulting in enhanced bone response and osseointegration.
Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases playa central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. In previous data, disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, cyclooxygenase (COX)-2 expression, and prostaglandin $E_2$ production in chondrocytes cultured on plastic or during cartilage explants culture. In this study, we investigated the effects of the actin cytoskeleton architecture on MMP-2 expression and dedifferentiation by CD in rabbit articular chondrocytes. Rabbit articular chondrocytes were prepared from cartilage slices of 2-weeks-old New Zealand white rabbits by enzymatic digestion. CD was used as a disruptor of actin cytoskeleton. In this experiments measuring CD dose response, primary chondrocytes were treated with various concentrations of CD for 24h. The actin disruption was determined by immunostaining. MMP-2 expression levels were determined by immunoblot analysis and Reverse transcriptase-Polymerase chain reaction (RT-PCR) and MMP-2 activity was determined by gelatin zymography. We found that cell morphological change and up-regulation of MMP-2 expression by CD as determined via immunostaining, gelatin zymography and immunoblotting. Moreover, CD induced MMP-2 transcription was detected by RT-PCR. Also, CD-induced type II collagen expression was inhibited by MMP-2 inhibitor I treatment. Our results indicate that CD up-regulated MMP-2 activation causes dedifferentiation of articular chondrocyte.
Kang, T. Y.;Yin, X. J.;Rho, G. J.;Lee, H.;Chae, Y. J.;Lee, H. J
Journal of Embryo Transfer
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v.15
no.2
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pp.167-173
/
2000
The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.
For the induction of arthropathy, 5% hydrogen peroxide($H_2O_2$) was injected for 5 weeks into the intraarticular space of the New Zealand white rabbits to damage articular cartilage. Alginic acid of low molecular weight (2%) made from macromolecular alginate treated with enzyme was administered into articular space at the dose of 5 mg/kg twice a week for 3 and 6 weeks using 1 ml syringe and 26 G needle. Saline was injected for the control. Tissues surrounding the articulation were obtained for the measurements of superoxide dismutase(SOD) activity as a major antioxidant enzyme and malondialdehyde (MDA) as a lipid peroxidation level. Histopathologic examination on the surface of articular cartilage was carried out. Data showed that injection of hydrogen peroxide for 5 weeks had led to the induction of free radical damage and of articular cartilage change as confirmed by microscopic observation. The application of hydrogen peroxide caused a gradual increase in the SODs and MDA. These patterns were similar after 3 and 6 weeks of alginate treatment. Furthermore, microscopic examinations revealed that hydrogen peroxide caused flaking, fibrillation, fissuring, denudation, and hypocellularity in the articular surfaces. In conclusion, lipid peroxidation was demonstrated in the articular cartilage by the administration of hydrogen peroxide in the rabbit model. This lipid peroxidation could be caused by oxygen free radicals. The histologic and enzymatic correlations on lipid peroxidation in the articulation have provided a better understanding of arthropathy. It is possible to take advantage of these findings to evaluate effective alginate dosage more efficiently.
Background: Caveolin, a family of integral membrane proteins are a principal component of caveolae membranes. In this study, we investigated the effect of p38 kinase on differentiation and on inflammatory responses in sodium nitroprusside (SNP)-treated chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. SNP was used as a nitric oxide (NO) donor. In this experiments measuring SNP dose response, primary chondrocytes were treated with various concentrations of SNP for 24h. The time course of the SNP response was determined by incubating cells with 1mM SNP for the indicated time period $(0{\sim}24h)$. The cyclooxygenase-2 (COX-2) and type II collagen expression levels were determined by immunoblot analysis, and prostaglandin $E_2\;(PGE_2)$ assay was used to measure the COX-2 activity. The tyrosine phosphorylation of caveolin-1 was determined by immunoblot analysis and immunostaining. Results: SNP treatment stimulated tyrosine phosphorylation of caveolin-1 and activation of p38 kinase. SNP additionally caused dedifferentiation and inflammatory response. We showed previously that SNP treatment stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase with SB203580 reduced caveolin-1 tyrosine phosphorylation and COX-2 expression but enhanced dedifferentiation, whereas inhibition of ERK with PD98059 did not affect caveolin-1 tyrosine phosphorylation levels, suggesting that ERK at least is not related to dedifferentiation and COX-2 expression through caveolin-1 tyrosine phosphorylation. Conclusion: Our results indicate that SNP in articular chondrocytes stimulates dedifferentiation and inflammatory response via p38 kinase signaling in association with caveolin-1 phosphorylation.
The principle of guided tissue regeneration (GTR), as applied to bone healing, is based on the prevention of connective tissue from entering the bony defect during the healing phase. This allows the slower bone producing cells to migrate into and reproduce bone within the defect. GTR has demonstrated a level of success in regenerating bone defect. Several types of membrane barrier have been utilized to apply this principle in bone regeneration. The purpose of this study was to evaluate whether improved bone regeneration can be achieved with different membrane barriers ($Gore-Tex^{TM}$membrane, $COLLACOTE^{(R)}$). In the 10 NewZealand white rabbits, full-thickness bone defects on three sites of each rabbit calvaria were made. Experimental group 1 was covered with $COLLACOTE^{(R)}$, and group 2 was covered with $Gore-Tex^{TM}$membrane. Macroscopic, microscopic examinations were made serially on 1, 2, 3, 6, 12 weeks after operation. The results were as follows : 1. Macroscopically, both of experimental group 1, 2 were filled with bone-like mass but the defects of experimental group 1 disclosed markedly thinner than the original bone. 2. Microscopically, the defect of experimental group 1, 2 was filled with bony trabeculae without infection and adverse reaction. But multinucleated giant cell infiltration around $COLLACOTE^{(R)}$ was seen till 6th week. 3. Resorption of $COLLACOTE^{(R)}$ started from 3rd week and it was completely resorped on the 12th week.
Chang, Jae Hoon;Park, Dae Hwan;Shin, Jeong Im;Ahn, Ki Young;Song, Chul Hong
Archives of Plastic Surgery
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v.33
no.1
/
pp.13-20
/
2006
Alginate, which is isolated from brown seaweed, is a bioabsorbable long chain polysaccharides, ${\beta}$-D-mannuronic acid and ${\alpha}$-L-guluronic acid. The authors produced alginates and alginate-colllagen as a disc form. Then, to evaluate the efficacy of alginate and alginate-collagen complex as a wound healing material, three full-thickness skin defects of 2 cm in diameter were made at the back of the New Zealand white rabbits. Three groups of dressing materials-alginate, alginate-collagen complex and vaseline gauze as control group - were applied on the wound and the results were evaluated grossly and histopathologically. The authors compared gross findings of sizes of healed wound, wound epithelization and wound contraction by tracing the remaining wound area at 5th, 10th, 15th, 20th, 25th day after wound introduction, and wound biopsy was performed at 3rd, 7th, 14th, 21st day, respectively. Alginate and alginate-collagen complex showed statistically higher percentage of wound contraction and wound healing compared to control group(p<0.05). Alginate-collagen complex showed statistically higher percentage of wound contraction, epithelization and wound healing compared to alginate alone. In conclusion, the result suggests that alginate has a good effect of wound healing and that alginate-collagen complex is more effective in wound healing than alginate alone.
Purpose: The aim of this study was to investigate the combined effects of physical and chemical surface factors on in vivo bone responses by comparing chemically modified hydrophilic sandblasted, large-grit, acid-etched (modSLA) and anodically oxidized hydrophobic implant surfaces. Methods: Five modSLA implants and five anodized implants were inserted into the tibiae of five New Zealand white rabbits (one implant for each tibia). The characteristics of each surface were determined using field emission scanning electron microscopy, energy dispersive spectroscopy, and confocal laser scanning microscopy before the installation. The experimental animals were sacrificed after 1 week of healing and histologic slides were prepared from the implant-tibial bone blocks removed from the animals. Histomorphometric analyses were performed on the light microscopic images, and bone-to-implant contact (BIC) and bone area (BA) ratios were measured. Nonparametric comparison tests were applied to find any significant differences (P<0.05) between the modSLA and anodized surfaces. Results: The roughness of the anodized surface was $1.22{\pm}0.17{\mu}m$ in Sa, which was within the optimal range of $1.0-2.0{\mu}m$ for a bone response. The modSLA surface was significantly rougher at $2.53{\pm}0.07{\mu}m$ in Sa. However, the modSLA implant had significantly higher BIC than the anodized implant (P=0.02). Furthermore, BA ratios did not significantly differ between the two implants, although the anodized implant had a higher mean value of BA (P>0.05). Conclusions: Within the limitations of this study, the hydrophilicity of the modSLA surface may have a stronger effect on in vivo bone healing than optimal surface roughness and surface chemistry of the anodized surface.
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