• Molecules and Cells
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• 제41권12호
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• pp.1016-1023
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• 2018

Regenerative orthopedics needs significant devices to transplant human stem cells into damaged tissue and encourage automatic growth into replacements suitable for the human skeleton. Soft biomaterials have similarities in mechanical, structural and architectural properties to natural extracellular matrix (ECM), but often lack essential ECM molecules and signals. Here we engineer mineralized polysaccharide beads to transform MSCs into osteogenic cells and osteoid tissue for transplantation. Bone morphogenic proteins (BMP-2) and indispensable ECM proteins both directed differentiation inside alginate beads. Laminin and collagen IV basement membrane matrix proteins fixed and organized MSCs onto the alginate matrix, and BMP-2 drove differentiation, osteoid tissue self-assembly, and small-scale mineralization. Augmentation of alginate is necessary, and we showed that a few rationally selected small proteins from the basement membrane (BM) compartment of the ECM were sufficient to up-regulate cell expression of Runx-2 and osteocalcin for osteoid formation, resulting in Alizarin red-positive mineral nodules. More significantly, nested BMP-2 and BM beads added to a non-union skull defect, self-generated osteoid expressing osteopontin (OPN) and osteocalcin (OCN) in a chain along the defect, at only four weeks, establishing a framework for complete regeneration expected in 6 and 12 weeks. Alginate beads are beneficial surgical devices for transplanting therapeutic cells in programmed (by the ECM components and alginate-chitosan properties) reaction environments ideal for promoting bone tissue.

• The Journal of Advanced Prosthodontics
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• 제5권2호
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• pp.167-171
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• 2013

PURPOSE. The purpose of this case series was to evaluate the effect of guided bone regeneration using demineralized allogenic bone matrix with calcium sulfate. MATERIALS AND METHODS. Guided bone regeneration using Demineralized Allogenic Bone Matrix with Calcium Sulfate ($AlloMatrix^{TM}$, Wright. USA) was performed at the time of implant placement from February 2010 to April 2010. At the time of the second surgery, clinical evaluation of bone healing and histologic evaluation were performed. The study included 10 patients, and 23 implants were placed. The extent of bony defects around implants was determined by measuring the horizontal and vertical bone defects using a periodontal probe from the mesial, distal, buccal, and lingual sides and calculating the mean and standard deviation of these measurements. Wedge-shaped tissue samples were obtained from 3 patients and histologic examination was performed. RESULTS. In clinical evaluation, it was observed that horizontal bone defects were completely healed with new bones, and in the vertical bone defect area, 15.1% of the original defect area remained. In 3 patients, histological tests were performed, and 16.7-41.7% new bone formation was confirmed. Bone graft materials slowly underwent resorption over time. CONCLUSION. $AlloMatrix^{TM}$ is an allograft material that can be readily manipulated. It does not require the use of barrier membranes, and good bone regeneration can be achieved with time.

• International Journal of Oral Biology
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• 제36권3호
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• pp.117-122
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• 2011

Endothelial cells are a vital constituent of most mammalian organs and are required to maintain the integrity of these tissues. These cells also play a major role in angiogenesis, inflammatory reactions, and in the regulation of thrombosis. Angiogenesis facilitates pulp formation and produces the vessels which are essential for the maintenance of tooth homeostasis. These vessels can also be used in bone and tissue regeneration, and in surgical procedures to place implants or to remove cancerous tissue. Furthermore, endothelial cell regeneration is the most critical component of the tooth generation process. The aim of the present study was to stimulate endothelial regeneration at a site of acute cyclophosphamide (CP)-induced endothelial injury by treatment with human umbilical cord-derived endothelial/mesenchymal stem cells (hEPCs). We randomly assigned 16 to 20-week-old female NOD/SCID mice into three separate groups, a hEPC ($1{\times}10^5$ cells) transplanted, 300mg/kg CP treated and saline (control) group. The mice were sacrificed on days 5 and 10 and blood was collected via the abdominal aorta for analysis. The alanine transaminase (ALT), aspartate aminotransferase (AST), serum alkaline phosphatase (s-ALP), and albumin (ALB) levels were then evaluated. Tissue sections from the livers and kidneys were stained with hematoxylin and eosin (HE) for microscopic analysis and were subjected to immunohistochemistry to evaluate any changes in the endothelial layer. CP treatment caused a weight reduction after one day. The kidney/body weight ratio increased in the hEPC treated animals compared with the CP only group at 10 days. Moreover, hEPC treatment resulted in reduced s-ALP, AST, ALT levels compared with the CP only group at 10 days. The CP only animals further showed endothelial injuries at five days which were recovered by hEPC treatment at 10 days. The number of CD31-positive cells was increased by hEPC treatment at both 5 and 10 days. In conclusion, the CP-induced disruption of endothelial cells is recovered by hEPC treatment, indicating that hEPC transplantation has potential benefits in the treatment of endothelial damage.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권2호
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• pp.97-106
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• 2010

Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권4호
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• pp.340-349
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• 2007

Free flap transplantation with microvascular anastomosis has been successfully performed by development of surgical technique, materials and postoperative monitoring equipments of flap. But success rate of microvascular anastomosis is influenced by various factors, and failure rate is about 5-10%. The most influential factor for success rate is surgical technique and other factors that influence failure of microvascular anastomosis are ischemic time of free flap, thrombus formation of anastomosis region and vascular spasm. In this study, vascular patency and thrombus formation in experimental micro-venous anastomosis, and endothelial repair were observed with histologic analysis, scanning electron microscopy, transmission electron microscopic examination. The results were obtained as follows: 1. In vascular patency test in 30 minute and 7 days after micro-venous anastomosis with heparin irrigation, all of 12 anastomosis site were good vascular patency. 2. In thrombus formation in 2 weeks group(Experimental I), 2 site of 6 cases were observed thrombus, and in 4 weeks group(Experimental II), 1 site of 6 cases were observed thrombus. 3. In histologic examination, normal vein(Control Group) showed continued internal elastic lamina, well formed thick smooth muscle layer and connective tissue. The group of 2 weeks after microvenous anastomosis(Experimental I) showd locally recovered internal lamina, discontinued internal lamina, disorganized smooth muscle cells and granulation tissue around suture silk. In the group of 4 weeks after micro-venous anastomosis(Experimental II), anastomosis site showed almostly continued internal lamina, disorganized smooth muscle cells and cicartrized tissue around suture silk. 4. In scanning electron microscope examination in 2 weeks(Experimental I) after micro-venous anastomosis, mesh fibrin formation showed near to endothelial cells, and in 4 weeks after micro-venous anastomosis(EXperimental II), numerous blood cells and fibrin mesh formation was seen associated with irregular endothelial cell arrangement. 5. In transmission electron microscope examination in 2 weeks after micro-venous anastomosis(Experimental I), irregular arrangement of smooth muscle cells was seen adjacent to collagenized tissue around suture silk. In 4 weeks after micro-venous anastomosis(Experimental II), denuded venous wall composed of relatively well arranged smooth muscle cells was covered by endothelial cells, but fibroblast cells and foreign body giant cells near to suture silk was remained. From the results obtained in this study, results of good vascular patiency and anti-thrombotic effect of heparin were obtained as a local irrigation solution, and repair of venous endothelial cell was observed in 2 weeks after micro-venous anastomosis.

• Journal of Periodontal and Implant Science
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• 제37권2호
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• pp.237-249
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• 2007

This study was performed to evaluate the effect of membrane exposure on new bone formation when guided bone regeneration with perforated titanium membrane on atrophic alveolar ridge. The present study attempted to establish a GBR model for four adult beagle dog premolar. Intra-marrow penetration defects were created on the alveolar ridge(twelve weeks after extraction) on the mandibular premolar teeth in the beagle dogs. Space providing perforated titanium membrane with various graft material were implanted to provide for GBR. The graft material were demineralized bovine bone(DBB), Irradiated cancellous bone(ICB) and demineralized human bone powder(DFDB). The gingival flap were advanced to cover the membranes and sutured. Seven sites experienced wound failure within 2-3weeks postsurgery resulting in membrane exposure. The animals were euthanized at 4 weeks postsurgery for histologic and histometric analysis. The results of this study were as follows: 1. There was little new bone formation at 4 weeks postsurgery. irrespectively of membrane exposure. 2. There was significant relationship between membrane exposure and bone graft resorption(P<0.05), but no relation between membrane exposure and infiltrated connective tissue. 3. There was much bone graft resorption on DFDB than ICB and DBB. 4. The less exposure was on the perforated titanium membrane, the more dense infiltrated connective tissue was filled under the membrane when grafted with ICB and DBB. but there was no relationship between the rate of membrane exposure and the percentage of infiltrated connective tissue area and no relationship between the percentage of the area in the infiltrated connective tissue and in the residual bone graft. Within the above results, bone formation may be inhibited when membrane was exposed and ICB and DBB were more effective than DFDB as a bone graft material when guided bone regeneration.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권3호
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• pp.188-195
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• 2002

The purpose of this investigation was to evaluate the relationship between the hydration time of demineralized freeze-dried bone (DFDB) and early new bone formation in rat calvarial defects filled with DFDB. Rats (n = 43) were divided into 4 experimental groups. Standard, transosseous circular defects of the calvaria were made midparietally. In experimental group 1, the defect was grafted immediately after soaking the DFDB. In experimental group 2, the defects were grafted with DFDB after soaking the DFDB for 10 minutes. In experimental groups 3 and 4, the defects were filled after soaking the DFDB for 30 and 60 minutes, respectively. Graft sites were analyzed histologically after healing periods of 1, 2, or 4 weeks. Each group showed similar bone regeneration at each time point by histological analysis. The results of this study were as follows: 1. After 1 week, a significant amount of inflammation, granulation tissue, and edema were found. A small amount of bone was seen, but the amount of bone did not differ between groups. 2. After 2 weeks, a small amount of new bone formation and DFDB resorption were observed. 3. After 4 weeks, a greater amount of new bone formation was observed. The greatest amount of bone formation occurred in experimental group 4 after 4 weeks. We conclude that the hydration time of DFDB does not affect new bone formation and that it is very important to control inflammation in bone grafting.

• Journal of Periodontal and Implant Science
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• 제50권6호
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• pp.406-417
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• 2020

Purpose: This study investigated whether the placement of ribose cross-linked collagen (RCLC) membranes without primary soft tissue closure predictably resulted in sufficient alveolar ridge preservation in contained and non-contained extraction sockets. Methods: Membranes were positioned across extraction sockets, undermining full-thickness flaps, and the gingival margins were fixed by double-interrupted sutures without crossed horizontal mattress sutures for 1 week. In non-contained sockets, a bone substitute was used to support the membrane within the bony envelope. Radiographs and clinical images obtained 4 months later were analyzed by ImageJ software using non-parametric tests. Results: In 18 patients, 20 extraction sockets healed uneventfully and all sites received standard-diameter implants (4.1, 4.8, or 5.0 mm) without additional bone augmentation. Soft tissues and the muco-gingival border were well maintained. A retrospective analysis of X-rays and clinical photographs showed non-significant shrinkage in the vertical and horizontal dimensions (P=0.575 and P=0.444, respectively). The new bone contained vital bone cells embedded in mineralized tissues. Conclusions: Within the limitations of this pilot study, open healing of RCLC membranes may result in sufficient bone volume for implant placement without additional bone augmentation in contained and non-contained extraction sockets.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제17권4호
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• pp.317-330
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• 1995

The purpose of this study is to observe the stimulant effect of the isolated nerve segment on the peripheral nerve regeneration using silicone tube in rats. Sprague-Dawley female albino rats were used as experimental animals and nerve defects were made by resection of 14 mm-long segment of right sciatic nerves. In control group, the defects were bridged with the silicone tubes. In experimental group, the defects were bridged with the same manner and additionally, 2mm-long nerve segments were inserted at the center of the silicone tubes. 3 months later, the regenerated nerve tissue within the silicone tubes were examined by histologic study. Axonal diameters and numbers of axons were measured and all data were analyzed by using SAS package program. The results obtained were as follows : 1. The experimental group was increased than the control group in general diameters of the regenerated nerve fiber.(P<0.05) 2. The diameters of axons were increased in the experimental group compare to the control group.(P<0.05) 3. The numbers of axons were increased in the experimental group compare to the control group.(P<0.05)

• Maxillofacial Plastic and Reconstructive Surgery
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• 제33권1호
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• pp.66-72
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• 2011

A severely vertical resorbed ridge is a significant challenge in implant dentistry. To solve this problem, several augmentation techniques, such as guided bone regeneration (GBR), onlay bone grafts, distraction osteogenesis, and ridge splitting techniques, have been proposed and used for several years. Among these methods, vertical ridge augmentation using guided bone regeneration aims to build space and guide osteoblasts to this space to promote osteogenesis. The aim of guided bone regeneration is to maintain and stabilize the space and block the proliferation of adjacent soft tissue. In our hospital, we encountered a case of a woman in her forties with an atrophied mandible, who underwent implant surgery in the right mandible. Titanium reinforced Gore-Tex (TRG) was used to augment the mandible and titanium mesh was used in the left mandible. Favorable results were obtained. This report compares the two methods and reviews the relevant literature.