• Journal of Periodontal and Implant Science
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• v.40 no.1
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• pp.11-18
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• 2010

Purpose: Significant interest has emerged in the design of cell scaffolds that incorporate peptide sequences that correspond to known signaling domains in extracellular matrix and bone morphogenetic protein. The purpose of this study was to evaluate the bone regenerative effects of the synthetic peptide in a critical-size rat calvarial defect model. Methods: Eight millimeter diameter standardized, circular, transosseus defects created on the cranium of forty rats were implanted with synthetic peptide, collagen, or both synthetic peptide and collagen. No material was was implanted the control group. The healing of each group was evaluated histologically and histomorphometrically after 2- and 8-week healing intervals. Results: Surgical implantation of the synthetic peptide and collagen resulted in enhanced local bone formation at both 2 and 8 weeks compared to the control group. When the experimental groups were compared to each other, they showed a similar pattern of bone formation. The defect closure and new bone area were significantly different in synthetic peptide and collagen group at 8 weeks. Conclusions: Concerning the advantages of biomaterials, synthetic peptide can be an effective biomaterial for damaged periodontal regeneration.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.1
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• pp.9-15
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• 2010

Aim of the study: As an injectable scaffold, MPEG-PCL diblock copolymer was applied in bone tissue engineering. In vivo bone formation was evaluated by soft X-ray, histology based on the rat calvarial critical size defect model. Materials and Methods: New bone formation was evaluated with MPEG-PCL diblock copolymer in rat calvarial critical size bone defect. No graft was served as control. 4, 8 weeks after implantation, gross evidence of bone regeneration was evaluated by histology and soft X-ray analysis. Results: The improved and effective bone regeneration was achieved with the BMP-2 and osteoblasts loaded MPEG-PCL diblock copolymer. Conclusion: It was confirmed that MPEG-PCL temperature sensitive hydrogels was useful as an injectable scaffold in bone regeneration.

• Journal of Nutrition and Health
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• v.28 no.8
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• pp.737-748
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• 1995

Osteoblast(OBL) cells were isolated from ICR Swiss neonatal mouse calvarial tissues and cultured in a CO2 incubator with minimum essential medium (MEM) containing 0.25g BSA. The cells were cultured for 7 days and were treated with bovine parathyroid hormone (bPTH, 1-34) and calcitonin(CT). Enzyme activities related to mineral metabolism and other biochemical actions within the bone cells including protein phosphorylation were investigated. In other experiments using cultured calvarial bone tissues, hormones were treated for 24, 48, 72 or 96 hours. The activities of $\beta$-glucuronidase enzymes involved in bone collagen synthesis and mineral deposits were increased by 8% with bPTH and were inhibited with CT treatment, while those were 67% increase treated with bPTH and CT together. On the other hand, alkaline phophatase(AP) activities were inhibited by PTH hormone at all the time courses observed. Protein phosphorylation reaction in OBL was mediated by bPTH, cAMP and ionized Ca. Phosphorylation was observed in different cell fractions including homogenate, membrane and cytosol. The number of proteins phosphorylated by PTH, cAMP, and Ca were 10, 5, and 9, respectively. Most of the protein kinases(PKs) were existed in cytosolic compartment. In membrane fractions, two bPTH-dependent-PKs (70K, 50K Da) were observed of which 70K Da protein was also Ca-dependent. Most of the cAMP-dependent PKs were regulated via bPTH. 70K, 50K, 5K, 19K, 16K, 10.5K phosphoproteins regulated by Ca share the same pathways as those by bPTH-dependent proteins. Ca seems to regulate PK activities differently from cAMP.

• BMB Reports
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• v.50 no.2
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• pp.97-102
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• 2017

Patients with inflammatory bone disease or cancer exhibit an increased risk of fractures and delayed bone healing. The S100A4 protein is a member of the calcium-binding S100 protein family, which is abundantly expressed in inflammatory diseases and cancers. We investigated the effects of extracellular S100A4 on osteoblasts, which are cells responsible for bone formation. Treating primary calvarial osteoblasts with recombinant S100A4 resulted in matrix mineralization reductions. The expression of osteoblast marker genes including osteocalcin and osterix was also suppressed. Interestingly, S100A4 stimulated the nuclear factor-kappaB (NF-${\kappa}B$) signaling pathway in osteoblasts. More importantly, the ex vivo organ culture of mouse calvariae with recombinant S100A4 decreased the expression levels of osteocalcin, supporting the results of our in vitro experiments. This suggests that extracellular S100A4 is important for the regulation of bone formation by activating the NF-${\kappa}B$ signaling pathway in osteoblasts.

• Journal of Periodontal and Implant Science
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• v.34 no.4
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• pp.733-746
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• 2004

The present study aimed to examine the effects of topical application of alendronate with a collagen membrane on the healing of the calvarial defect in rats, which has a good experimental design for the healing of tissue destruction, To study the effect of alendronate on bone healing, the collagen membrane containing $200{\mu}g$ alendronate was inserted in the defects of the right side and collagen membrane treated with physiologic saline was inserted in the defects of the left side. After 1, 2 and 4 weeks, observation of histologic feature after H&E staining, cell counting after TRAP staining, and hardness measurement(Knoop) were performed. In histologic finding, similar features were shown for both test and control groups each week. In cell counting only the 1 week test groups showed significant reduction of TRAP(+)cells than control groups(p<0.01) and the control groups showed statistically significant difference for 1, 2, 4weeks(p<0.05). In hardness measurement, The 2 week test groups showed significant higher hardness than control groups.(p<0,05) and not 4 weeks. There was significant increase of hardness for both groups as time goes by.(p<0.0l) Therefore local application of alendronate with collagen membrane was somewhat effective in reducing osteoclastic activity and increasing hardness in the early stage of healing. Further investigation concerning the actual effect of alendronate for bony healing will be necessary to apply the clinical cases

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.489-500
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• 2002

The purpose of this study was to evaluate newly fabricated tricalcium phosphate(TCP)/chitosan microgranuls as bone substitutes. TCP/chitosan microgranules were fabricated by dropping TCP-chitosan suspension into the NaOH/ethanol solution. The size of microgranules could be controllable via airflow rate. PDGF-BB was loaded into the fabricated granules via freeze-drying methods(300 ng/20 mg). To evaluate cell proliferation, cultured osteoblasts cell lines(MC3T3-El) was dropped on the BioOss(R), chitosan microgranules, TCP/chitosan microgranules and cultured for 1, 7 , 14, and 28 days. Scanning electron microscopic observation was done after 7 days of culture and light microscopic examination was done after 28 days of culture. PDGF-BB release from the microgranules was tested. Rabbit calvarial defects(8 mm in diameter) were formed and chitosan, TCP/chitosan, PDGF-TCP/chitosan microgranules, and BioGran(R) were grafted to test the ability of new bone formation. At SEM view, the size of prepared microgranules was 250-1000 um and TCP powders were observed at the surface of TCP/chitosan microgranules. TCP powders gave roughness to the granules and this might help the attachment of osteoblasts. The pores formed between microgranules might be able to allow new bone ingrowth and vascularization. There were no significant differences in cell number among BioOss(R) and two microgranules at 28 day. Light and scanning electron microscopic examination showed that seeded osteoblastic cells were well attached to TCP/chitosan microgranules and proliferated in a multi-layer. PDGF-BB released from TCP/chitosan microgranules was at therapeutic concentration for at least 1 week. In rabbit calvarial defect models, PDGF-TCP/chitosan microgranules grafted sites showed thicker bone trabeculae pattern and faster bone maturation than others. These results suggested that the TCP/chitosan microgranules showed the potential as bone substitutes.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.5
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• pp.440-453
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• 2005

Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

• Journal of Korean Neurosurgical Society
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• v.58 no.3
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• pp.167-174
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• 2015

Objective : This study investigates the role of a burr hole and calvarial bone marrow-derived stem cells (BMSCs) in a transient ischemic brain injury model in the rat and postulates a possible mechanism for the efficacy of multiple cranial burr hole (MCBH) surgery in moyamoya disease (MMD). Methods : Twenty Sprague-Dawley rats (250 g, male) were divided into four groups : normal control group (n=5), burr hole group (n=5), ischemia group (n=5), and ischemia+burr hole group (n=5). Focal ischemia was induced by the transient middle cerebral artery occlusion (MCAO). At one week after the ischemic injury, a 2 mm-sized cranial burr hole with small cortical incision was made on the ipsilateral (left) parietal area. Bromodeoxyuridine (BrdU, 50 mg/kg) was injected intraperitoneally, 2 times a day for 6 days after the burr hole trephination. At one week after the burr hole trephination, brains were harvested. Immunohistochemical stainings for BrdU, CD34, VEGF, and Doublecortin and Nestin were done. Results : In the ischemia+burr hole group, BrdU (+), CD34 (+), and Doublecortin (+) cells were found in the cortical incision site below the burr hole. A number of cells with Nestin (+) or VEGF (+) were found in the cerebral parenchyma around the cortical incision site. In the other groups, BrdU (+), CD34 (+), Doublecortin (+), and Nestin (+) cells were not detected in the corresponding area. These findings suggest that BrdU (+) and CD34 (+) cells are bone marrow-derived stem cells, which may be derived from the calvarial bone marrow through the burr hole. The existence of CD34 (+) and VEGF (+) cells indicates increased angiogenesis, while the existence of Doublecortin (+), Nestin (+) cells indicates increased neurogenesis. Conclusion : Based on these findings, the BMSCs through burr holes seem to play an important role for the therapeutic effect of the MCBH surgery in MMD.

• Journal of Marine Bioscience and Biotechnology
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• v.7 no.2
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• pp.58-70
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• 2015

Injectable RGD-bioconjugated Mussel Adhesive Proteins (RGD-MAPs) composite hydroxypropyl methylcellulose (HPMC) hydrogels provide local periodontal tissue for bone filling in periodontal surgery. Previously we developed a novel type of injectable self-supported hydrogel (2 mg/ml of RGD-MAPs/HPMC) based porcine nano hydroxyapatite (MPH) for dental graft, which could good handling property, biodegradation or biocompatibility with the hydrogel disassembly and provided efficient cell adhesion activity and no inflammatory responses. Herein, the aim of this work was to evaluate bone formation following implantation of MPH and collagen membrane in rabbit calvarial defects. Eight male New Zealand rabbits were used and four circular calvarial defects were created on each animal. Defects were filled with different graft materials: 1) collagen membrane, 2) collagen membrane with MPH, 3) collagen membrane with bovine bone hydroxyapatite (BBH), and 4) control. The animals were sacrificed after 2 and 8 weeks of healing periods for histologic analysis. Both sites receiving MPH and BBH showed statistically increased augmented volume and new bone formation (p < 0.05). However, there was no statistical difference in new bone formation between the MPH, BBH and collagen membrane group at all healing periods. Within the limits of this study, collagen membrane with MPH was an effective material for bone formation and space maintaining in rabbit calvarial defects.

• Archives of Plastic Surgery
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• v.35 no.6
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• pp.631-636
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• 2008

Purpose: Adipose tissue-derived stem cells(ADSC) has an osteoconductive potential and demineralized bone matrix(DBM) is an osteoinductive material. A combination of DBM and ADSC wound probably create osteoinductive properties. The purpose of this study is to determine the effect of the combination of DBM and ADSC mixture on healing of rat calvarial defect. Methods: Thirty adult male Sprague-Dawley rats were randomized into 3 groups(n=10) as 1) Control, 2) DBM alone, 3) DBM with ADSC mixture. DBM with ADSC mixture group has had a 3-day preculture of ADSC from groin fat pad. An 6 mm critical size circular calvarial defect was made in each rat. Defect was implanted with DBM alone or DBM with ADSC mixture. Control defect was left unfilled. 6 and 12 weeks after the implantation, the rats were sacrificed and the defects were evaluated by histomorphometric and radiographical studies. Results: Histomorphometric analysis revealed that DBM with ADSC mixture group showed significantly higher bone formation than DBM alone group(p<0.05). Although radiographs from DBM alone group and DBM with ADSC group revealed similar diffuse radiopaque spots dispersed throughout the defect. Densitometric analysis of calvarial defect revealed DBM with ADSC mixture group significantly higher bone formation than DBM alone(p<0.05). There was correlation of densitometry with new bone formation(Spearman's correlation of coefficient=0.804, 6 weeks, 0.802, 12 weeks). Conclusion: The DBM with ADSC mixture group showed the best healing response and the osteoinductive properties of DBM were accelerated with ADSC mixture. It will be clinically applicable that DBM and ADSC mixture in plastic and reconstructive surgery, such as alveolar cleft and congenital facial deformities that bone graft should be required.