• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.1
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• pp.1-11
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• 2012

Purpose: The purpose of this study was to evaluate the bone regeneration capacity of silk fibroin (SF) when combined with beta tricalcium phosphate (${\beta}$-tricalcium phosphate [TCP]) and rh-bone morphogenetic protein (BMP) in vivo by micro-computed tomography (CT), soft x-ray, and histological analysis. Methods: A total of 56 critical size defects formed by a trephine bur made on 28 adult female Spague-Dawley rats were used for this study and the defect size was 5.0 mm in diameter. The defects were transplanted with (1) no graft material (raw defect), (2) autogenous bone, (3) SF ($10{\mu}g$), (4) SF-BMP ($10{\mu}g$, $0.8{\mu}g$ each), and (5) SF+${\beta}$-TCP ($10{\mu}g$). At 4 and 8 weeks after operation, the experimental animals were sacrificed. Samples were evaluated with soft x-ray, histological examinations and 3-dimensional micro-CT analysis. Results: In the 3-dimensional micro-CT evaluation, bone volume and bone surface data were higher in the SF-BMP ($12.8{\pm}1.5$, $138.6{\pm}45.0$ each) (P<0.05) and SF-TCP ($12.3{\pm}1.5$, $144.9{\pm}30.9$ each) group than in the SF group ($6.1{\pm}3.3$, $77.2{\pm}37.3$ each) (P<0.05), except for the autogenous group ($15.0{\pm}3.0$, $190.7{\pm}41.4$ each) at 4 weeks. At 8 weeks, SF-BMP ($16.8{\pm}3.5$, $173.9{\pm}34.2$ each) still revealed higher (P<0.05) bone volum and surface, but SF-TCP ($11.3{\pm}1.5$, $1132.9{\pm}52.1$ each) (P=0.5, P=0.2) revealed the same or lower amount compared with the SF group ($13.8{\pm}2.7$, $127.5{\pm}44.8$ each). The % of bone area determined by radiodensity was higher in the SF-TCP ($31.4{\pm}9.1%$) and SF-BMP ($36.2{\pm}16.2%$) groups than in the SF ($19.0{\pm}10.4$) group at the period of 4 weeks. Also, in the histological evaluation, the SF-BMP group revealed lower inflammation reaction, lower foreign body reaction and higher bone healing than the SF group at postoperative 4 weeks and 8 weeks. The SF-TCP group revealed lower inflammation at 4 weeks, but accordingly, as the TCP membrane was absorbed, inflammatory and foreign body reaction are increased at 8 weeks. Conclusion: The current study provides evidence that the silk fibrin can be used as an effective grafted material for tissue engineering bone generation through a combination of growth factor or surface treatment.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.347-361
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• 1997

The present study investigates the effects of DFDB graft combined with Calcium sulfate membrane on the periodontal wound healing in dehiscence defects of dogs. Following the initiation of general anesthesia by I.V. administration of 30mg/kg of pentobarbital, first premolar was extracted and full-thickness flap was elevated from the second to the fourth premolar. The portion of premolars coronal to the alveolar crest was removed and mesial and distal roots separated to produce single rooted teeth. Exposed root canals were sealed with Caviton and covered completely with flaps sutured. Following the healing period of 12 weeks, the surgical sites were uncovered and $4{\times}4mm$ dehiscence defects were surgically created. Those defects with DFDB graft combined with Calcium sulfate membrane following root planing, were designated as test sites and those with flap surgery-only were designated as controls. 1. No foreign-body reaction or inflammation were observed in either groups. Calcium sulfate was completely resorbed in the test sites. 2. Significantly greater amounts of new cementum was observed in test sites compared with the controls. Significant amounts of functionally orientated collagens were observed in the test sites. 3. New bone formation was observed in significantly greater amounts in test sites. The results suggest that combined graft of DFDB and calcium sulfate is extremely biocompatible with a potential for new bone and cementum formation, and functional alignment of periodontal ligaments.

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.249-262
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• 1997

The present study investigates the effects of calcium sulfate graft on the periodontal healing in intrabony periodontal defects of dogs. Following the general anesthesia with 30mg/kg pentobarbital injected intravenously, the first premolar was extracted and full-thickness periodontal flap was elevated from the second premolar to the fourth premolar. The portion of premolars coronal to the alveolar crest was removed and mesial and distal roots were separated. Exposed root canals were sealed with Caviton and covered completely with flaps sutured. Following the healing period of 12 weeks, the surgical sited were uncovered and $4{\times}4mm$ intrabony defects were surgically created. Those defects with calcium sulfate graft following the root planing was designated as the test sites and those with flap surgery-only were designated as control sites. The animals were sacrificed after 8 weeks and the healing was histologically analyzed. The results were as follows. 1. No foreign body reaction or inflammation were observed in either groups. Calcium sulfate was completely resorbed in the test sites. 2. New cementum was observed coronal to the notch in both groups. Connective tissue fibers were oriented parallel to the root surface in the controls. Connective tissues were formed in large amount in the sites. 3. Test sites showed marked amount of new bone formation while the control sites showed minimal bone gain. 4. Root resorption was observed in coronal portions of th control Sites. The results suggest that calcium sulfate is a biocompatible graft material with a potential for new bone and cementum formation.

• KSBB Journal
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• v.16 no.3
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• pp.302-306
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• 2001

The possibility of biomaterials prepared from natural polymer as a skin substitute was evaluated by measuring biocompatibility. Biodegradable films were prepared by solution blend method in the weight ratio of chitosan for the purpose of useful biomaterials. These films were inserted in the back of rat and their biodegradability was investigated by the film weight and hematology as a function of time for the biotransformation. The result of rat test showed that medium, high viscosity chitosan induced some suspects of inbiocompatibililty in the tissue by foreign body reaction 48 and 72 hours after implantation. Also, we prepared the official burn ointment which is made by low viscosity chitosan. This burn ointment was covered on the skin wound of artificial burn and their effect of healing was investigated by the evaluation of the naked eye and hematological change as a function of time. The result of rats test showed that burn ointments made from chitosan was effective reduction of inflammation than negative group.

• Archives of Reconstructive Microsurgery
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• v.3 no.1
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• pp.45-53
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• 1994

Suture microvascular anastomosis is time-consuming and tedious and demands long and continuous training. Techinique of anastomosis of microvessel was presented interrupted suture and continuous suture. Recently the unilink instrument system is created as a fast and simple method to achieve high patency rates without long and continuous training in the anastomosis of small vessels. The author experimentally studied the femoral artery of 20 mice(0.5-1.0mm, av. 0.7mm), the femoral vein of 20 mice(0.8-1.6mm, av. 1.2mm) after anastomosis with interrupted suture in 20 cases and continuous sutre in 20 cases. For the unilink apparatus we used the carotid arteries of 15 cases in 14 rabbits(1.0-1.6mm, av. 1.3mm) and facial veins of 12 cases in 14 rabbits(0.9mm-2.2mm, av. 1.5mm). A total of 27 arterial and venous anastomoses were performed. We examined the postoperative patency at immediate, 2 weeks, and 8 weeks. The results were as followings, 1. In the arterial anastomosis the rate of patency was 90%(18/20) in interrupted suture, 90%(18/20) in continuous suture and 93%(13/15) in unilink apparatus. In the venous anastomosis the rate of patency was 90%(18/20) in interrupted suture, 80%(16/20) in continuous suture and 100%(9/9) in unilink apparatus. 2. The mean time for completion of the arterial anastomosis were 12.2 minutes in interrupted suture group, 10.3 minutes in continouous suture group and 8.5 minutes in unillnk apparatus group. The mean time for completion of the venous anastomosis were 13.6 minutes in interrupted suture group, 11.0 minutes in continuous suture group and 6.2 minutes in unilink apparatus group. 3. At the histological examination of suture group, hyperplastic reaction of middle layer and subintimal hyperplasia were observed. In unilink apparatus group, the endothelium layer was continued and the thickness of vessel wall was decreased due to moderate atrophy of the media and mild degree of nonspecific chronic inflammation were seen around the unilink apparatus. 4. No significants was noticied in foreign body reaction among the interrupted, continuous and unilink apparatus group. 5. A case of the arterial anastomosis was released with acting out at 15 minutes after operation. 6. The important factors in the technical problems were accurate apposition of the cut vessel edges in suture group and the proper selection of the ring size and optimal fitting between two rings in unilink apparatus group. Even though the outer diamater of vessel in suture group was different from that in unilink apparatus group the unilink method provides a very safe, fast, and simple way to perform microvascular anastomoses especially in anastomosis of vein. But howerver suture was needed in vessels below 1 mm outer diamater. In that situation continuous suture was benefit than the interrupted suture in operation time.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.5
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• pp.366-374
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• 2010

Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.633-648
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• 1994

Synthetic bone graft materials have been used for the regeneration of periodontal tissue lost due to periodontal disease, but the limitations of these materials had prompted the use of composite grafts. Among those, a composite graft of calcium carbonate(CC) and calcium sulfate(CS) is one of those materials that has not been studied extensively. CC, which is extracted from a natural coral, is known to possess osteoconductive property. SC can play an adjunctive role in the regeneration of bone tissue, and has shown good resorbability and biocompatibility. This study was conducted in order to investigate the effects of CC and CS composite graft to the regeneration of bone in the intrabony defects of dogs. 3-wall intrabony defects ub size of $4mm{\times}4mm{\times}4mm$ were created in the alveolar bone in the premolar areas. Then those defects that were treated with root planning only were designated as control, while the experimental group 1 and 2 each received the CC and CS composite grafts in the ratio of 8 : 2 and 5 : 5 the animals were sacrificed after 8weeks and the specimens were histologically analyzed. The results were as follows ; 1. No inflammation or foreign body reaction were observed in all subjects. CS has not been seen due to complete resorption, and resorption pattern of CC was observed. 2. Significant differences(p<0.05) in new cementum formation were observed between control($1.42{\pm}0.64mm$) and experimental groups(group 1 ; $2.53{\pm}0.94mm$, group 2 ; $2.23{\pm}0.96mm$) but the difference between the two experimental groups was not significant. 3. Significant differences(p<0.01) in new bone formation were observed between control($0.59{\pm}0.55mm$) and experimental groups(group 1 ; $2.27{\pm}0.61mm$, group 2 ; $2.05{\pm}0.56mm$) but the difference between the two experimental groups was not significant. 4. The extent of apical epithelial migration has shown no significant difference between control($1.18{\pm}1.24mm$) and experimental groups(group 1 ; $0.51{\pm}0.54mm$, group 2 ; $0.73{\pm}0.70mm$). 5. The extent of bone formation was generally limited to the extent of cementum formation for all groups, and significant correlation was found in the amount of bone formation and cementum formation in experimental group 1.(Co.=0.86, p<0.01) These results suggest that the composite graft of CC and CS is biocomplatible and effective in the new bone and new cementum formations. In the case of 3-wall intrabony defects of dogs, the composite ratio of 8 : 2 and 5 : 5 had shown no significant differences in the healing.

• Toxicological Research
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• v.35 no.1
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• pp.45-63
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• 2019

In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ($BMM{\Phi}$) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.

• Journal of Dental Rehabilitation and Applied Science
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• v.40 no.1
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• pp.1-12
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• 2024

Purpose: To histologically evaluate the effects of three calcium silicate-based sealers on rat mandible tissue. Materials and Methods: Rats were randomly divided as follows: A group that sacrificed immediately after cavity preparation, a group that sacrificed two weeks after cavity preparation, a group that sacrificed two weeks after CeraSeal (CS), AH Plus Bioceramic (AHB), or One-Fil (OF) sealer injection, respectively. After tissue processing for all groups, the bone tissue area (%) and the number of osteoclasts in and around the cavity were measured under a microscope. The results of each group were compared and statistical analysis was performed using one-way ANOVA and Tukey's test. Results: The formation of bone tissue and the presence of osteoclasts in the cavity were observed in the group that sacrificed two weeks after cavity preparation and the group sacrificed two weeks after AHB sealer injection, and these groups showed significantly higher average bone tissue area (%) than the other groups. In the other groups, no inflammation or foreign body reaction occurred in the cavity, and no osteoclasts were observed. Conclusion: All calcium silicate-based sealers used in this study showed a favorable bone tissue response when injected into the rat mandible. In particular, higher bone formation in the cavity was observed in AHB.