• Journal of Biomedical Engineering Research
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• v.43 no.1
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• pp.61-70
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• 2022

Natural and synthetic forms of calcium phosphate cement (CPC) have been widely used in bone repair and augmentation. The major challenge of injectable CPC is to deliver the cells without cell death in order to regenerate new bone. The study objective was to investigate for the potential of stem cell-laden gelatin fibers containing injectable, nanocrystalline CPC to function as a delivery system. Gelatin noddle fiber method was developed to delivered cells into nCPC. Experimental groups were prepared by mixing cells with nCPC, mixing cell-laden gelatin fibers with nCPC and mixing cell-laden gelatin fibers containing BMP-2 with nCPC. Media diffusion test was conducted after dissolving the gelatin fibers. SEM examined the generated channels and delivered cell morphology. Fibers mixed with nCPC showed physical setting and hardening within 20 min after injection and showed good shape maintenances. The gelatin fibers mixed nCPC group had several vacant channels generated from the dissolved gelatin. Particularly, proliferation and attachment of the cells were observed inside of the channels. While live cells were not observed in the cell mixed nCPC group, cells delivered with the gelatin fibers into the nCPC showed good viability and increased DNA content with culture. Cell-laden gelatin fiber was a novel method for cell delivery into nCPC without cell damages. Results also indicated the osteogenic differentiation of gelatin fiber delivered cells. We suggest that the cell-laden gelatin fibers mixed with nCPC can be used as an injectable cell delivery vehicle and the addition of BMP-2 to enhances osteogenesis.

• Ocean and Polar Research
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• v.44 no.1
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• pp.29-38
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• 2022

Halophytes are plants that live in harsh environments in coastal regions and are known for their diverse chemical compositions. Limonium tetragonum, a halophyte endemic to Korean shores, is known for its bioactive compounds and is utilized in folk medicine. In this study L. tetragonum extract (LHE) was used to determine and evaluate its anti-osteoporotic properties. Pre-adipocyte and pre-osteoblasts were induced to differentiate along with LHE treatment, and their differentiation was evaluated using differentiation markers. LHE treatment decreased lipid accumulation in 3T3-L1 preadipocytes during adipogenesis. Results indicated that the LHE treatment also decreased the levels of key adipogenic transcription factors: PPARγ, SREBP1c, and C/EBPα. Enhancing osteoblastogenesis by LHE treatment was confirmed in osteoblastogenesis-induced MC3T3-E1 pre-osteoblasts. Cells treated with LHE resulted in increased calcification and alkaline phosphatase (ALP) activity compared with osteoblasts without LHE treatment. Pro-osteogenic and anti-adipogenic effects were also confirmed in D1 murine mesenchymal stromal cells which are capable of differentiation into both adipocytes and osteoblasts. LHE hindered adipogenesis and enhanced osteoblastogenesis in D1 MSCs in a similar fashion. In conclusion, L. tetragonum is believed to possess the potential to be utilized as a nutraceutical ingredient against osteoporotic conditions.

• Journal of Periodontal and Implant Science
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• v.52 no.4
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• pp.312-324
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• 2022

Purpose: The role of hard-type crosslinked hyaluronic acid (HA) with particulate bone substitutes in bone regeneration for combined inlay-onlay grafts has not been fully investigated. We aimed to evaluate the effect of hard-type crosslinked HA used with bone substitute in terms of new bone formation and space maintenance. Methods: A 15-mm-diameter round defect was formed in the calvaria of 30 New Zealand White rabbits. All animals were randomly assigned to 1 of 3 groups: the control group (spontaneous healing without material, n=10), the biphasic calcium phosphate (BCP) graft group (BCP, n=10), and the BCP graft with HA group (BCP/HA, n=10). The animals were evaluated 4 and 12 weeks after surgery. Half of the animals from each group were sacrificed at 4 and 12 weeks after surgery. Samples were evaluated using micro-computed tomography, histology, and histomorphometry. Results: The BCP group showed higher bone volume/tissue volume (BV/TV) values than the control and BCP/HA groups at both 4 and 12 weeks. The BCP and BCP/HA groups showed higher bone surface/tissue volume (BS/TV) values than the control group at both 4 and 12 weeks. The BCP group showed higher BS/TV values than the control and BCP/HA groups at both 4 and 12 weeks. No statistically significant difference in newly formed bone was found among the 3 groups at 4 weeks. The BCP group showed significantly higher new bone formation than the BCP/HA group at 12 weeks. Conclusions: Hard-type crosslinked HA did not show a positive effect on new bone formation and space maintenance. The negative effect of hard-type crosslinked HA may be due to the physical properties of HA that impede osteogenic potential.

• Journal of Korean Dental Science
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• v.17 no.2
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• pp.64-74
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• 2024

This case report presents the long-term radiographic outcomes of a novel approach for simultaneous lateral augmentation and implant surgery. A 60-year-old male patient who required tooth extraction of the maxillary central and lateral incisors due to trauma visited the clinic. After tooth extraction, severe horizontal and vertical deficiencies occurred owing to atrophy of the alveolar ridge, and a simultaneous guided bone regeneration (GBR) procedure was planned along with the installation of two implants. In the present case, a modification of the conventional 'sandwich technique' was used by placing the mixture of autogenous bone chips and xenografts at the outermost layer to maximize the osteogenic potential at the coronal part of augmentation while applying solely xenografts at the inner layer. To enhance volumetric stability, an autogenous block of periosteum harvested from the maxillary tuberosity was incorporated between the two layers. Cone-beam computed tomography was performed at baseline and 3 years after the surgery to compare radiographic outcomes. Dehiscence after fixture installation was successfully observed at the re-entry of the surgery site. Three years after the surgery, average horizontal bone gains of 6.11 mm and 4.12 mm were observed in the maxillary central and lateral incisor areas, respectively. Healthy peri-implant mucosa and well maintained marginal bone levels were observed 8 years after the surgery, meeting the criteria for implant success. The findings of this case suggest that a substantial amount of horizontal bone gain can be obtained with a layered approach using autogenous bone materials and xenografts, highlighting the advantages of incorporating autogenous blocks into the simultaneous GBR procedure.

• Journal of Life Science
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• v.28 no.12
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• pp.1448-1454
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• 2018

Insects have been investigated as a novel source of food and biomaterial in several recent studies. However, their osteoblastogenic cell activity has not been sufficiently researched and so, to investigate the potential of this natural material for promoting osteoblastogenesis, we studied the activity of Locusta migratoria ethanol extract (LME) on MG-63 pre-osteoblast cells. The cytotoxicity and proliferation effects of LME on MG-63 cells were measured by MTS assay, and there was no cytotoxicity up to $1,000{\mu}g/ml$. With LME treatment of 500 and $1,000{\mu}g/ml$ for 48 hr, cell proliferation increased to 105% and 116% versus control, respectively. The osteoblastogenic activity of the LME was measured through alkaline phosphatase (ALP) staining at three and five days. As a result, both 500 and $1,000{\mu}g/ml$ LME concentrations were seen to increase ALP activity by more than three times compared with control at three and five days. In addition, the expression level of the osteogenic markers ALP and RUNX2 was markedly increased after LME treatment. These results demonstrate that Locusta migratoria ethanol extract promotes osteoblastogenesis as evidenced by the increased osteogenic markers and suggest that LME may be a potential agent for bone formation and osteoporosis prevention.

• Journal of the korean academy of Pediatric Dentistry
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• v.42 no.4
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• pp.291-301
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• 2015

This study compared the in vitro cell viability and differentiation potentials of human deciduous dental pulp cells (DPCs) on mineral trioxide aggregate (MTA)-like products (ProRoot MTA, RetroMTA and Endocem Zr). The experimental materials were prepared as circular discs, which were used to test the effects of the materials on the viability of human DPCs when placed in direct and indirect contact. Furthermore, the pH of the extracted materials was recorded, and their effect on cell differentiation potential was evaluated by evaluating the alkaline phosphatase (ALP) activity and Alizarin Red S staining of DPCs incubated with the test materials. In direct contact, the cell viability of human DPCs was higher with ProRoot MTA and RetroMTA than with Endocem Zr. However, when in indirect contact, the cell viability of human DPCs was generally higher in Endocem Zr than in ProRoot MTA and Retro MTA. With respect to pH, the alkalinity was lower for Endocem Zr than for the other test materials. The ALP activities of the cells were not enhanced by any of the experimental materials. Alizarin Red S staining of the tested human DPCs revealed that their differentiation potential was lower than for cells incubated with osteogenic induction medium. While there were differences in the responses of the human DPCs to the test materials, all displayed degrees of cytotoxicity and were unable to enhance either the viability or differentiation of human DPCs. However, Endocem Zr exhibited better cell viability and was less alkaline than the other test materials.

• Journal of Microbiology and Biotechnology
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• v.32 no.8
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• pp.1017-1025
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• 2022

Bone homeostasis is regulated by constant remodeling through osteogenesis by osteoblasts and osteolysis by osteoclasts and osteoporosis can be provoked when this balance is broken. Present pharmaceutical treatments for osteoporosis have harmful side effects and thus, our goal was to develop therapeutics from intrisincally safe natural products. Fucoidan is a polysaccharide extracted from many species of brown seaweed, with valuable pharmaceutical activities. To intensify the effect of fucoidan on bone homeostasis, we hydrolyzed fucoidan using AMG, Pectinex and Viscozyme. Of these, fucoidan biotransformed by Pectinex (Fu/Pec) powerfully inhibited the induction of tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts differentiated from bone marrow macrophages (BMMs) by the receptor for activation of nuclear factor-κB ligand (RANKL). To investigate potential of lower molecular weight fucoidan it was separated into >300 kDa, 50-300 kDa, and <50 kDa Fu/Pec fractions by ultrafiltration system. The effects of these fractions on TRAP and alkaline phosphatase (ALP) activities were then examined in differentiated osteoclasts and MC3T3-E1 osteoblasts, respectively. Interestingly, 50-300 kDa Fu/Pec suppressed RANKL-induced osteoclasts differentiation from BMMs but did not synergistically enhance osteoblasts differentiation induced by osteogenic agents. In addition, this fraction inhibited the expressions of NFATc1, TRAP, OSCAR, and RANK, which are all key transcriptional factors involved in osteoclast differentiation, and those of Src, c-Fos and Mitf, as determined by RT-PCR. In conclusion, enzymatically low-molecularized 50-300 kDa Fu/Pec suppressed TRAP by downregulating RANKL-related signaling, contributing to the inhibition of osteoclasts differentiation, and represented a potential means of inducing bone remodeling in the background of osteoporosis.

• Archives of Reconstructive Microsurgery
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• v.9 no.2
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• pp.139-146
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• 2000

The aim of this study was to assess whether the functional regeneration of a lyophilized autografted cartilage could be improved by implanting a vascularized muscle flap into the medullary canal of autografted proximal humerus. A hemijoint reconstruction using a lyophilized osteochondral autograft in proximal humerus was done in 4 rabbits for control, and combined with an vascularized intramedullary muscle flap in another 4 rabbits for the experimental group. Graft healing and the repair process of osteochondral graft were followed by serial radiographs and histologic changes for 9 weeks after experiments. Each two rabbits in control and in experimental group on 5th and 9th week after implantation of hemijoint were sacrified. The results were as follows: 1. All of control and experimental froups on 5th week united solidly on osteotomized site radiologically, but their articular cartilages were destroyed more seriously in the control than that in experimental group with muscle flap on 5th and 9th week after experiment... 2. Histochemically, the cartilage surface are completely destroyed and revealed with severe osteoarthritic changes on all cartilage layers in control, but cartilaginous erosions are mild to moderate and their arthritic changes are also mild with somewhat regeneration of chondrocytes on deep layers more prominetly on 9th week of the experimental group. 3. The amount of collagen and protenized matrix which was determined by Masson-Trichrome stain was markedly decreased that means the weakness of bony strength and low osteogenic potential in lyophilized cartilage. These results suggest that an intramedullary vascularized muscle flap can improve the functional results of lyophilized osteochondral autograft by providing both increased vascularity and populations of mesenchymal cells to initiate new bone formation on osteotomized site as well as the regeneration of deep layers in articular cartilage. In clinical relevances, this lyophilized hemijoint autograft combined with an intramedullary vascularized muscle pedicle graft might be used very effectively for the treatment of malignant long bone tumors to preserve the joint functions, all or partly, and so to replace it with the artificial joint after tumor excision and hemijoint autograft.

• Journal of Periodontal and Implant Science
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• v.23 no.2
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• pp.315-330
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• 1993

The ultimate goal of periodontal treatment has been to facilitate regeneration of diseased periodontal tissues, destroyed by inflammatory periodontal disease. Various implant materials have been used to restore the alveolar bone defects. Of the various materials, porous replamineform hydroxyapatite (PHA) has good biocompatibility when placed in a bone tissue, and maintains alveolar ridge for a long period. Decalcified freeze dried bone(DFDB) has been widely used in alveolar bone defects because of its conformity and high osteogenic potential. The purpose of this study was to evaluate the effects of PHA and DFDB on the regeneration of the alveolar bone between fresh extraction sockets and periodontally involved extraction sockets. Experimental periodontitis was induced by the ligation of orthodontic elastic threads after surgically creating periodontal defects on the premolars on the right side of 2 adult dogs for 8 weeks. Following the extraction of each tooth, PHA and DFDB were inserted in the extraction sockets. In control group 1, PHA was inserted in the fresh extraction sockets, and in control group 2, DFDB was inserted. In experimental group 1, PHA was inserted in the periodontally involved extraction sockets, and in experimental group 2, DFDB was inserted. After 20 weeks, the specimens were prepared and stained with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows. 1. No inflammation associated with implant materials was evident in any of the groups. 2. DFDB was completely resorbed, PHA was remained in the extraction sockets in the control and experimental groups. 3. In control group 1 and experimental group 1, extraction sockets were not completely filled with new bone. However, original forms of alveolar crests were maintained in control group 2 and experimental group 2. 4. In control group 1 and exprimental group 1, PHA particles surrounded with many giant cells were well tolerated by the fibrous connective tissues in the coronal part of the socket, In the inferior part of the socket, PHA particles were incorporated into the new bone. In both control group 2 and experimental group 2, DFDB was replaced by newly remodeled bone. 5. No differences of degree of new bone formation were evident between control and experimental groups.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.363-377
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• 1997

The main goal of periodontal therapy is the regeneration of periodontal tissue which has been lost due to destructive periodontal diseases. Although conventional forms of periodontal therapy show sound clinical results, the healing results in long junctional epithelium. There have been numerous materials and surgical techniques developed for new attachment and bone regeneration. Bone grafts can be catagorized into: autografts, allografts, xenografts and bone substitutes. Synthetic bone substitute materials include hydroxyapatite, tricalcium phosphate, calcium carbonate, and Plaster of Paris. Calcium sulfate has found its use in dental practice for the last 30 years. Recent animal studies suggest that periodontal regeneration in 3 wall intrabony defect may be enhanced by the presence of calcium sulfate. And it is well known that 2 wall & 1 wall defect have less osteogenic potential, So we need to study the effect of calcium sulfate in 1 wall intrabony defect in dogs. The present study evaluates the effects of calcium sulfate on the epithelial migration, alveolar bone regeneration and cementum formation in intrabony defects of dogs. Four millimeter-deep one-wall intrabony defects were surgically created in the mesial aspect of anterior teeth and mesial & distal aspects of premolars. The test group received calcium sulfate grafts with a flap procedure. The control underwent flap procedure only. Histologic analysis following 8 weeks of healing revealed the following results: 1. The lengths of junctional epithelium were: 2.52mm in the control, and 1.89mm in the test group. There was no statistical significance between the two groups. 2. Alveolar bone formation were: 0.61mm in the control, and 1.88mm in the test group. There was a statistically significant difference between the two groups (p<0.05). 3. Cementum formations were: l.lmm in the control, and 2.46mm in the test group. There was a statistically significant difference between the two groups (p<0.05). 4. The length of CT adhesion were: O.97mm in the control, and 0.17mm in the test group. There was no statistically significant differences between the two groups These results suggest that the use of calcium sulfate in intrabony defects has little effect on junctional epithelium migration, but has significant effects on new bone and new cementum formations.