Background Composite grafts are frequently used for facial reconstruction. However, the unpredictability of the results and difficulties with large defects are disadvantages. Adipose-derived stem cells (ADSCs) express several cytokines, and increase the survival of random flaps and fat grafts owing to their angiogenic potential. Methods This study investigated composite graft survival after ADSC injection. Circular chondrocutaneous composite tissues, 2 cm in diameter, from 15 New Zealand white rabbits were used. Thirty ears were randomly divided into 3 groups. In the experimental groups (1 and 2), ADSCs were subcutaneously injected 7 days and immediately before the operation, respectively. Similarly, phosphate-buffered saline was injected in the control group just before surgery in the same manner as in group 2. In all groups, chondrocutaneous composite tissue was elevated, rotated 90 degrees, and repaired in its original position. Skin flow was assessed using laser Doppler 1, 3, 6, 9, and 12 days after surgery. At 1 and 12 days after surgery, the viable area was assessed using digital photography; the rabbits were euthanized, and immunohistochemical staining for CD31 was performed to assess neovascularization. Results The survival of composite grafts increased significantly with the injection of ADSCs (P<0.05). ADSC injection significantly improved neovascularization based on anti-CD31 immunohistochemical analysis and vascular endothelial growth factor expression (P<0.05) in both group 1 and group 2 compared to the control group. No statistically significant differences in graft survival, anti-CD31 neovascularization, or microcirculation were found between groups 1 and 2. Conclusions Treatment with ADSCs improved the composite graft survival, as confirmed by the survival area and histological evaluation. The differences according to the injection timing were not significant.
Background: Ischemic injury and the rejection process are the main reasons for graft failure in tracheal transplantation models. To enhance the acceptance, we investigated the influence of mesenchymal stem cells (MSCs) on tracheal allografts. Methods: Extracted tracheal grafts from New Zealand white rabbits were cryopreserved for 4 weeks and orthotopically transplanted (control group A, n=8). In group B (n=8), cyclosporin A (CsA, 10 mg/kg) was injected daily into the peritoneal cavity. In group C (n=8), MSCs ($1.0{\times}10^7$ cells/kg) from the same donor of the tracheal allograft, which had been pre-cultured for 4 weeks, were infused intravenously after transplantation. In group D (n=8), MSCs were infused and CsA was injected daily. Four weeks after transplantation, gross and histomorphological assessments were conducted for graft necrosis, measuring the cross-sectional area of the allograft, determining the degree of epithelization, lymphocytic infiltration, and vascular regeneration. Results: The morphologic integrity of the trachea was retained completely in all cases. The cross-sectional areas were decreased significantly in group A (p=0.018) and B (p=0.045). The degree of epithelization was enhanced (p=0.012) and the lymphocytic infiltration was decreased (p=0.048) significantly in group D compared to group A. The degree of vascular regeneration did not differ significantly in any of the groups. There were no significant correlations among epithelization, lymphocytic infiltration, and vascular regeneration. Conclusion: The administration of MSCs with concurrent injections of CsA enhanced and promoted epithelization and prevented lymphocytic infiltration in tracheal allografts, allowing for better acceptance of the allograft.
PURPOSE. The purpose of this study was to assess the effect of systemically administered oxytocin (OT) on the implant-bone interface by using histomorphometric analysis and the removal torque test. MATERIALS AND METHODS. A total of 10 adult, New Zealand white, female rabbits were used in this experiment. We placed 2 implants (CSM; CSM Implant, Daegu, South Korea) in each distal femoral metaphysis on both the right and left sides; the implants on both sides were placed 10 mm apart. In each rabbit, 1 implant was prepared for histomorphometric analysis and the other 3 were prepared for the removal torque test (RT). The animals received intramuscular injections of either saline (control group; 0.15 M NaCl) or OT (experimental group; $200{\mu}g/rabbit$). The injections were initiated on Day 3 following the implant surgery and were continued for 4 subsequent weeks; the injections were administered twice per day (at a 12-h interval), for 2 days per week. RESULTS. While no statistically significant difference was observed between the two groups (P=.787), the control group had stronger removal torque values. The serum OT concentration (ELISA value) was higher in the OT-treated group, although no statistically significant difference was found. Further, the histomorphometric parameter (bone-to-implant contact [BIC], inter-thread bone, and peri-implant bone) values were higher in the experimental group, but the differences were not significant. CONCLUSION. We postulate that OT supplementation via intramuscular injection weakly contributes to the bone response at the implant-bone interface in rabbits. Therefore, higher concentrations or more frequent administration of OT may be required for a greater bone response to the implant. Further studies analyzing these aspects are needed.
Choi, Eun Joo;Kang, Sang-Hoon;Kwon, Hyun-Jin;Cho, Sung-Won;Kim, Hyung Jun
Maxillofacial Plastic and Reconstructive Surgery
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제36권3호
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pp.94-102
/
2014
Purpose: This study aims to validate the effect of autoclaved autogenous bone (AAB), incorporating Escherichia coli-derived recombinant human bone morphogenetic protein-2 (ErhBMP-2), on critical-sized, segmental radius defects in rabbits. Delivery systems using absorbable collagen sponge (ACS) and fibrin glue (FG) were also evaluated. Methods: Radius defects were made in 12 New Zealand white rabbits. After autoclaving, the resected bone was reinserted and fixed. The animals were classified into three groups: only AAB reinserted (group 1, control), and AAB and ErhBMP-2 inserted using an ACS (group 2) or FG (group 3) as a carrier. Animals were sacrificed six or 12 weeks after surgery. Specimens were evaluated using radiology and histology. Results: Micro-computed tomography images showed the best bony union in group 2 at six and 12 weeks after operation. Quantitative analysis showed all indices except trabecular thickness were the highest in group 2 and the lowest in group 1 at twelve weeks. Histologic results showed the greatest bony union between AAB and radial bone at twelve weeks, indicating the highest degree of engraftment. Conclusion: ErhBMP-2 increases bony healing when applied on AAB graft sites. In addition, the ACS was reconfirmed as a useful delivery system for ErhBMP-2.
Shin, Seung-Woo;Pyo, Sung-Woon;Bae, Sun-Sook;Lee, Pil-Woo;Heo, Hyun-A;Lee, Won
Journal of Korean Dental Science
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제4권2호
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pp.39-45
/
2011
Purpose: The goal of this research is to find the role of collagen membrane, which can reduce physical damage, as a scaffold for possible alternative to the corticotomy which causes Regional Acceleratory Phenomenon (RAP). Materials and Methods: The experiments were carried out on 12 New Zealand white rabbits, approximately 3.5 kg in bodyweight. We made an incision on the skin of the mandibular border and applied 37% phosphoric acid and collagen membrane to the mandibular bone surface of the first group (experimental group), and only phosphoric acid to the second group (control group). After 3 days, 1 week, and 2 weeks, 4 rabbits each were sacrificed and specimens were obtained. Each specimen was stained by H&E and Tartrate-resistant acid phosphatase (TRAP), and histological changes were observed by light microscope. Results: The demineralization of the experimental group was weak compared to the control group. It also showed a gradual increase of demineralization (after 3 days, 1 week, and 2 weeks) and the control group showed more extensive demineralization than the experimental group. Conclusion: This study demonstrates the amount of demineralization as a result of using phosphoric acid, and as time went by, demineralization increased. The absorbable collagen membrane was used as a scaffold to increase bone demineralization effect and prevent dispersion to adjacent tissues, but rather the amount of bone demineralization decreased. Therefore, the role of collagen membrane as a scaffold for RAP was weak.
Statement of problem : The survival rate of wide diameter implants was lower than of 3.75-mm implants in some clinical researches. Purpose : The purpose of this study was to investigate the influence of implant diameter on the osseointegration of implants in the rabbit femoral condyle and tibial metaphyses by means of removal torque measurements and histomorphometric analysis. Material and Method : Ten adult New Zealand White rabbits were used in this study Two 3.75-mm diameter implants were inserted through one cortical layer in the tibial metaphyses and one 3.75-mm diameter implant was inserted in the femoral condyle. 5.0-mm diameter implants were inserted in the other leg in the same manner. A total of 60 implants (3.75-mm diameter implants:30 : 5.0-mm diameter implants:30) were installed. After a healing time of 4 and 12 weeks, the peak removal torque values required to shear off the implants were recorded. From the removal torque values (Ncm) obtained, the mean shear stress ($N/mm^2$) was calculated. And the percentage of direct bone-to-implant contact and the percentage of bone area inside the thread were measured by Kappa Image Base-metreo. The Student's t-test was undertaken for statistical analysis (p<0.05). Results : The removal torque value of 5.0-mm diameter implants was higher than of 3.75-mm diameter implants (p<0.05). The difference of shear stress value between 3.75-mm and 5.0-mm diameter implants was not statistically significant (p>0.05). The percentage direct bone-to-implant contact had no statistical difference between two groups (p>0.05). The percentage of bone area inside the thread had no statistical difference between two groups (p>0.05). Conclusion It is concluded that the quality f osseointegration is not influenced by increasing implant diameter.
Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.
Hypercholesterolemia indirectly increases the risk of arterial and venous thrombosis by enhancing the ability of platelets to aggregate. Yacon (Smallanthus sonchifolius) is composed of fructooligosaccharides, proteins, minerals and phenolic compounds, and has potential benefits for the management of diabetes. This study investigated whether the consumption of yacon in the diet inhibits platelet aggregation under hypercholesterolemic conditions. Male New Zealand white rabbits were fed one of five dietary interventions: a normal control diet, 0.5% cholesterol diet, 0.5% cholesterol diet+a low dose of yacon (0.5 g/kg body weight given orally each day), 0.5% cholesterol diet+a high dose of yacon (2.5 g/kg body weight given orally each day), or a 0.5% cholesterol diet+lovastatin (2 mg/kg body weight given orally each day). After 8 weeks, blood was collected to measure the amount of collagen- and thrombin-induced platelets present. Yacon inhibited the platelet aggregation induced by low doses of agonists (0.5 ${\mu}g/mL$ collagen and 0.02 units/ml thrombin) in a concentration-dependent manner. In addition, yacon concentration-dependently inhibited collagen-induced arachidonic acid liberation. Moreover, n-hexane, chloroform and ethyl acetate fractions showed a marked and selective inhibition of the platelet aggregation induced by collagen, again in a dose-dependent manner. These fractions, especially that of chloroform, significantly suppressed platelet aggregation. The results of this study demonstrate that when yacon is added to a cholesterol-enriched diet, cholesterol-induced platelet aggregation returns to control levels. This may also be beneficial in preventing atherosclerosis and reducing risk factors for coronary artery disease and stroke.
Yoo, Hyokyung;Yoon, Taekeun;Bae, Hahn-Sol;Kang, Min-Suk;Kim, Byung Jun
대한두개안면성형외과학회지
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제22권5호
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pp.260-267
/
2021
Background: Elastic ear cartilage is a good source of tissue for support or augmentation in plastic and reconstructive surgery. However, the amount of ear cartilage is limited and excessive use of cartilage can cause deformation of the auricular framework. This animal study investigated the potential of periosteal chondrogenesis in an ear cartilage defect model. Methods: Twelve New Zealand white rabbits were used in the present study. Four ear cartilage defects were created in both ears of each rabbit, between the central artery and marginal veins. The defects were covered with perichondrium (group 1), periosteum taken from the calvarium (group 2), or periosteum taken from the tibia (group 3). No coverage was performed in a control group (group 4). All animals were sacrificed 6 weeks later, and the ratio of neo-cartilage to defect size was measured. Results: Significant chondrogenesis occurred only in group 1 (cartilage regeneration ratio: mean±standard deviation, 0.97±0.60), whereas the cartilage regeneration ratio was substantially lower in group 2 (0.10±0.11), group 3 (0.08±0.09), and group 4 (0.08±0.14) (p= 0.004). Instead of chondrogenesis, osteogenesis was observed in the periosteal graft groups. No statistically significant differences were found in the amount of osteogenesis or chondrogenesis between groups 2 and 3. Group 4 showed fibrous tissue accumulation in the defect area. Conclusion: Periosteal grafts showed weak chondrogenic potential in an ear cartilage defect model of rabbits; instead, they exhibited osteogenesis, irrespective of their embryological origin.
Purpose: Increased bone regeneration has been achieved through the use of stem cells in combination with graft material. However, the survival of transplanted stem cells remains a major concern. The purpose of this study was to evaluate the viability of transplanted mesenchymal stem cells (MSCs) at an early time point (24 hours) based on the type and form of the scaffold used, including type I collagen membrane and synthetic bone. Methods: The stem cells were obtained from the periosteum of the otherwise healthy dental patients. Four symmetrical circular defects measuring 6 mm in diameter were made in New Zealand white rabbits using a trephine drill. The defects were grafted with 1) synthetic bone (${\beta}$-tricalcium phosphate/hydroxyapatite [${\beta}-TCP/HA$]) and $1{\times}10^5MSCs$, 2) collagen membrane and $1{\times}10^5MSCs$, 3) ${\beta}-TCP/HA+collagen$ membrane and $1{\times}10^5MSCs$, or 4) ${\beta}-TCP/HA$, a chipped collagen membrane and $1{\times}10^5MSCs$. Cellular viability and the cell migration rate were analyzed. Results: Cells were easily separated from the collagen membrane, but not from synthetic bone. The number of stem cells attached to synthetic bone in groups 1, 3, and 4 seemed to be similar. Cellular viability in group 2 was significantly higher than in the other groups (P<0.05). The cell migration rate was highest in group 2, but this difference was not statistically significant (P>0.05). Conclusions: This study showed that stem cells can be applied when a membrane is used as a scaffold under no or minimal pressure. When space maintenance is needed, stem cells can be loaded onto synthetic bone with a chipped membrane to enhance the survival rate.
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