Purpose: This study was performed to evaluate the effect of various graft materials used with a titanium cap on the ability of new bone formation in the rabbit calvarium. Materials and Methods: A total of 32 sites of artificial bony defects were prepared on the calvaria of sixteen rabbits by using a trephine bur 8 mm in diameter. Each rabbit had two defect sites. 0.2 mm deep grooves were formed on the calvaria of sixteen rabbits by using a trephine bur 8 mm in diameter for the fixation of a titanium cap. The treatments were performed respectively as follows: without any graft for the control group (n=8), autogenous iliac bone graft for experimental group 1 (n=8), alloplastic bone graft ($SynthoGraft^{(R)}$, USA) for experimental group 2 (n=8), and xenogenic bone graft ($NuOss^{(R)}$, USA) for experimental group 3 (n=8). After the treatments, a titanium cap (8 mm in diameter, 4 mm high, and 0.2 mm thick) was fixed into the groove. At the third and sixth postoperative weeks, rabbits in each group were sacrificed for histological analysis. Results: 1. In gross examination, the surgical sites showed no signs of inflammation or wound dehiscence, and semicircular-shaped bone remodeling was shown both in the experimental and control groups. 2. In histological analysis, the control group at the third week showed bone remodeling along the inner surface of the cap and at the contact region of the calvarium without any specific infiltration of inflammation tissue. Also, there was no soft tissue infiltration. Bone remodeling was observed around the grafted bone and along the inner surface of the titanium cap in experimental group 1, 2, and 3. 3. Histologically, all groups at the sixth week showed the increased area of bone remodeling and maturation compared to those at the third week. In experimental group 2, the grafted bone was partially absorbed by multi nucleated giant cells and new bone was formed by osteoblasts. In group 3, however, resorption of the grafted bone was not observed. 4. Autogenous bone at the third and sixth week showed the most powerful ability of new bone formation. The size of newly formed bone was in decreasing order by autogenous, alloplastic, and heterogenous bone graft. There was no statistically significant difference among autogenous, alloplastic, and heterogenous bones(p>0.05). Summary: This result suggests that autogenous bone is the best choice for new bone formation, but when autogenous bone graft is in limited availability, alloplastic and xenogenic bone graft also can be an alternative bone graft material to use with a suitably guided membrane.
A synoptic survery of chemical characteristics in the last Sea of Korea was carried out at the 11 stations near Ullungdo in November, 1994 on board R/V Tam-Yang. On the basis of the vortical distribution patterns of temperature, salinity and dissolved oxygen, water masses in the study area are divided into five groups; 1) Tsushima Surface Water (TSW), 2) Tsushima Middle Water (TMW), 3) East Sea Intermediate Water (ESIW), 4) last Sea Proper Water (ESPW), 5) Mixed Water (MW). In the vertical profiles of nutrients, the concentrations were very low in the surface layer and increased rapidly near the thermocline. There was a slight decrease in the ESIW and the concentrations were constant with the depth below 300m except dissolved silicate which still increased with depth. Relatively high value of Si/P ratio (25.2) in ESPW, whick is the oldest water mass, suggests that Si is regenerating more slowly compared to other nutrients. The relatively high value of N/P ratio (18.6) in the surface layer might be related to high vertical eddy diffusivity $(K_z)$ of $1.19\;cm^{2}/sec$ and high nitrate upward flux of $103.7\;{\mu}g-at/m^{2}/hr$, compared to the values reported in other areas. Apparent Oxygen Utilization (AOU) was very low in the surface layer and increased in the TMW, but there was a slight decrease in the ESIW. The highest value of AOU occurred in the ESPW. The slpoe of P/AOU was 0.50. The study on the relationship between water masses and nutrient distribution patterns is important in understanding the regeneration processes of nutrients in the polar region of the last Sea.
Min, Sung Ran;Ko, Suk Min;Lyu, Jae Il;Park, Ji Hyun;Yi, So Young;Lee, In-Ha;Kim, Hyun Sook;Kim, Tae Il;Choi, Pil Son;Jeong, Won-Joong;Kim, Suk Weon;Kim, Jonghyun;Liu, Jang R.
Journal of Plant Biotechnology
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v.42
no.3
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pp.180-185
/
2015
Leaf discs from 'Yeobong' and 'Maehyang' strawberry plants were used as explants for transformation. The Agrobacterium tumefaciens strain EHA105 harboring the monellin gene under the control of the CaMV 35S promoter was used in co-cultivation experiments. The frequencies of callus formation and plant regeneration from leaf explants after co-cultivation in 'Yeobong' were higher than those of 'Maehyang'. These transgenic plants showed normal growth patterns and flowering. PCR and Southern hybridization confirmed that 1 to 2 copies of the monellin gene were integrated into genome of the transgenic strawberry plants. Northern blot analysis confirm that the transcripts were expressed in transgenic strawberry plants. Although long-term subcultured transgenic strawberry plants showed a phenomenon to escape the transgene, the transformation system established in this study provides new opportunities for genetic improvement of strawberry plants.
thermochemical methane reforming consisting of two steps on Cu/Fe/Zr mixed oxide media was carried out using a fixed bed infrared reactor. In the first step, the metal oxide was reduced with methane to produce CO, $H_2$ and the reduced metal oxide in the temperature of 1173 K. In the second step, the reduced metal oxide was re-oxidized with steam to produce $H_2$ and the metal oxide in the temperature of 973 K. The reaction characteristics on the added amounts of Cu in Cu/Fe/Zr mixed oxide media and the cyclic tests were evaluated. With the increase of the added amount of Cu in Cu/Fe/Zr mixed oxide media, the conversion of $CH_4$, the selectivity of $CO_2$ and the $H_2/CO$ molar ratio were increased, while the selectivity of CO was decreased in the first step. On the other hand, the evolved amount of $H_2$ was decreased with increasing the added amount of Cu in the second step. The $Cu_xFe_{3-x}O_4/ZrO_2$ medium added with Cu of x = 0.7 showed good regeneration properties in the 10th cyclic tests indicating that the medium had high durability. In addition, the gasification of the deposited carbon in the water splitting step was promoted with the addition of Cu in the media.
The purpose of this study was to evaluate the pulp tissue reaction to direct pulp capping of mechanically exposed beagle dogs' pulp with several capping materials. A total of 36 teeth of 2 healthy beagle dongs were used. The mechanically exposed pulps were capped with one of the followings: (1) Mineral Trioxide Aggregate (MTA: $ProRoot^{(R)}$ MTA. Dentsply, Tulsa, USA), (2) Clearfil SE Bond (Dentin adhesive system: Kuraray, Osaka, Japan), (3) Ultra-Blend (Photo-polymerized Calcium hydroxide: Ultradent, South Jordan, USA), (4) Dycal (Quick setting Calcium hydroxide: LD Caulk Co., Milford, USA) at 7, 30, and 90 days before sacrificing. The cavities were restored with Z350 flowable composite resin (3M ESPE, St. Paul. MN, USA). After the beagle dogs were sacrificed, the extracted teeth were fixed, decalcified, prepared for histological examination and stained with HE stain. The pulpal tissue responses to direct pulp capping materials were assessed. In MTA calcium hydroxide, and photo-polymerized calcium hydroxide groups, initial mild inflammatory cell infiltration, newly formed odontoblast-like cell layer and hard tissue bridge formation were observed. Compared with dentin adhesive system, these materials were biocompatible and good for pulp tissue regeneration. In dentin adhesive system group, severe inflammatory cell infiltration, pulp tissue degeneration and pulp tissue necrosis were observed. It seemed evident that application of dentin adhesive system in direct pulp capping of beagle dog teeth cannot lead to acceptable repair of the pulp tissue with dentine bridge formation.
Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.
In dentistry, bony defects can be formed by cyst, tumor, inflammation, trauma and surgery in maxilla and mandible. If the overlying soft tissue invades and preoccupies the jaw bony defects, regenerated bony tissue same as adjacent bone can not replace whole space of the defects, thus preventing osteogenesis from occurring. Guided bone regeneration(GBR) is based on the prevention of overlying soft tissue from entering the bony defect during the initial healing periods. E-polytetrafluoroethylene(e-PTFE) is one of an effective and widely used barrier membrane for GBR, but it has the disadvantages such as surgical removal and high price. To overcome such disadvantages of e-PTFE, many investigators have proposed various absorbable barrier membranes. Inexpensive oxidized cellulose($Surgicel^{(R)}$) membrane was shown to have potential for use as an absorbable barrier membrane for regenerative procedure and it would not require surgical removal. The purpose of this study is to investigate the absorption periods of oxidized cellulose at the implant site and usefulness as a mechanical barrier, preventing the ingrowth of the overlying soft tissue into the bony defects. Two bony defects were made in each tibia of a dog using drill and one defect covered with oxidized cellulose and the other covered with periosteum directly as control. The experimental animals were sacrificed at 1st-7th, 10th, 14th, 21th, 28th day postoperatively, Inspection of the specimens was done to evaluate gross changes. Specimens were examined histopathologically by hematoxylin-eosin and Masson's trichrome staining under light microscope. The results were as follows : 1. There was no significant differences of inflammatory reaction between the experimental and the control group. 2. The resorption of oxidized cellulose was almost completed within 14th day. 3. Histologically, bone formation in the experimental group was somewhat more than that of the control group at 10th, 14th, 21th and 28th day postoperatively. The bone forming pattern of the experimental group was more regular than that of the control group. 4. There was no evidence of soft tissue invasion into the bony defect in the experimental group. In conclusion, oxidized cellulose membrane might be used as an alternative absorbable barrier membrane to prevent overlying soft tissue invasion into the bony defects.
Background: Hyaluronic acid (HA) has been applied as a primary biomaterial for temporary soft tissue augmentation and as a carrier for cells and the delivery of growth factors to promote tissue regeneration. Although HA derivatives are the most versatile soft tissue fillers on the market, they are resorbed early, within 3 to 12 months. To overcome their short duration, they can be combined with cells or growth factors. The purpose of this study was to investigate the stimulating effects of human fibroblasts and basic fibroblast growth factors (bFGF) on collagen synthesis during soft tissue augmentation by HA hydrogels and to compare these with the effects of a commercial HA derivative (Restylane®). Methods: The hydrogel group included four conditions. The first condition consisted of hydrogel (H) alone as a negative control, and the other three conditions were bFGF-containing hydrogel (HB), human fibroblast-containing hydrogel (HF), and human fibroblast/bFGF-containing hydrogel (HBF). In the Restylane® group (HGF), the hydrogel was replaced with Restylane® (R, RB, RF, RBF). The gels were implanted subdermally into the back of each nude mouse at four separate sites. Twelve nude mice were used for the hydrogel (n = 6) and Restylane® groups (n = 6). The specimens were harvested 8 weeks after implantation and assessed histomorphometrically, and collagen synthesis was evaluated by RT-PCR. Results: The hydrogel group showed good biocompatibility with the surrounding tissues and stimulated the formation of a fibrous matrix. HBF and HF showed significantly higher soft tissue synthesis compared to H (p < 0.05), and human collagen type I was well expressed in HB, HF, and HBF; HBF showed the strongest expression. The Restylane® filler was surrounded by a fibrous capsule without any soft tissue infiltration from the neighboring tissue, and collagen synthesis within the Restylane® filler could not be observed, even though no inflammatory reactions were observed. Conclusion: This study revealed that HA-based hydrogel alone or hydrogel combined with fibroblasts and/or bFGF can be effectively used for soft tissue augmentation.
Park, Shin-Young;Kim, Kyoung-Hwa;Koo, Ki-Tae;Lee, Kang-Woon;Lee, Yong-Moo;Chung, Chong-Pyoung;Seol, Yang-Jo
Journal of Periodontal and Implant Science
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v.41
no.5
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pp.218-226
/
2011
Purpose: Micro-computed tomography (micro-CT) has been widely used in the evaluation of regenerated bone tissue but the reliability of micro-CT has not yet been established. This study evaluated the correlation between histomorphometric analysis and micro-CT analysis in performing new bone formation measurement. Methods: Critical-size calvarial defects were created using a 8 mm trephine bur in a total of 24 Sprague-Dawley rats, and collagen gel mixed with autogenous rat bone marrow stromal cells (BMSCs) or autogenous rat BMSCs transduced by adenovirus containing bone morphogenic protein-2 (BMP-2) genes was loaded into the defect site. In the control group, collagen gel alone was loaded into the defect. After 2 and 4 weeks, the animals were euthanized and calvaria containing defects were harvested. Micro-CT analysis and histomorphometric analysis of each sample were accomplished and the statistical evaluation about the correlation between both analyses was performed. Results: New bone formation of the BMP-2 group was greater than that of the other groups at 2 and 4 weeks in both histomorphometric analysis and micro-CT analysis (P=0.026, P=0.034). Histomorphometric analysis of representative sections showed similar results to histomorphometric analysis with a mean value of 3 sections. Measurement of new bone formation was highly correlated between histomorphometric analysis and micro-CT analysis, especially at the low lower threshold level at 2 weeks (adjusted $r^2=0.907$, P<0.001). New bone formation of the BMP-2 group analyzed by micro-CT tended to decline sharply with an increasing lower threshold level, and it was statistically significant (P<0.001). Conclusions: Both histomorphometric analysis and micro-CT analysis were valid methods for measurement of the new bone in rat calvarial defects and the ability to detect the new bone in micro-CT analysis was highly influenced by the threshold level in the BMP-2 group at early stage.
This study compared the short-term(4 months) clinical results of regenerative therapy with bioabsorbable membranes($BioMesh^{(R)}$) and bone allograft for the treatment of periodontal(intrabony and furcation) defects in smokers and nonsmokers.(16 smokers) 32 subjects with 92 defects participated in the study(46 in smokers and 46 in non-smokers). This study also evaluated a bioresorbable barrier with and without decalcified freeze-dried bone allograft(DFDBA). The 92 periodontal defects were randomly treated with either the resorbable barrier alone or resorbable barrier in combination with DFDBA following thorough defect debridement and root preparation with tetracycline. Each patient received both types of treatment modalities. Clinical examinations(probing depth, gingival recession, clinical attachment level, plaque index and gingival index) were carried out immediately before and 4 months after surgery. Significant(p<0.001) gains in mean attachment level were observed for both smokers(2.93mm) and non-smokers(3.30mm) but there were not significant difference between two groups. Similarly, significant reductions in mean probing depthshowed for smokers(4.52mm) and non-smokers(4.26mm). However, when comparing gingival recession, smokers were found to exhibit significantly poorer treatment results(1.59mm vs 0.96mm, p<0.05). Using the split-mouth-design, no statistically significant difference between the two modalities could be detected with regard to pocket depth reduction, gingival recession, or attachment gain. These results illustrate that the attachment gain is better in the non-smoker and the best in the non-smoker with the combination therapy of resorbable barrier and DFDBA than with resorbable barrier alone but smoking had no significant effect on clinical treatment outcome, even though smokers show more significant gingival recession. In addition, both treatments, either resorbable barrier plus DFDBA or resorbable barrier alone, promoted significant resolution of periodontal defects but the addition of DFDBA with a bioabsorbable membrane appears to add no extra benefit to the only membrane treatment.
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