• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.6
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• pp.384-390
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• 2012

Purpose: The purpose of this study is to examine in vivo osteogenesis of cultured human periosteal-derived cells and polydioxanone/pluronic F127 scaffold. Methods: Two one-year-old miniature pigs were used in this study. $2{\times}10^6$ periosteal-derived cells in 1 mL medium were seeded by dropping the cell suspension into the polydioxanone/pluronic F127 scaffold. These cell-scaffold constructs were cultured in osteogenic Dulbecco's modified Eagle's medium for 7 days. Under general anesthesia with azaperone and tiletamine-zolazepam, the mandibular body and ramus of the pigs were exposed. Three bony defects were created. Polydioxanone/pluronic F127 scaffold with periosteal-derived cells and the scaffold only were implanted into each defect. Another defect was left empty. Twelve weeks after implantation, the animals were sacrificed. Results: New bone formation was clearly observed in the polydioxanone/pluronic F127 scaffold with periosteal-derived cells. Newly generated bone was also observed in the scaffold without periosteal-derived osteoblasts and empty defect, but was mostly limited to the periphery. Conclusion: These results suggest that cultured human periosteal-derived cells have good osteogenic capacity in a polydioxanone/pluronic F127 scaffold, which provides a proper environment for the osteoblastic differentiation of these cells.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.25 no.2
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• pp.423-435
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• 1995

The purpose of this study was to observe the effect of TGF-βl, which promotes differentiation and proliferation of osteoblasts, on bone regeneration. Experimental bone defects that measured 3 mm in diameter were created on the mandibles of guinea pig by removal of bone with the use of trephine burs. In one side of mandibular body, the experimental groups, bone defects were grafted with Biogran(Orthovita Co., U.S.A) and TGF-β1(R&D System Co., U.SA). In the remaining side of the mandiblar body, the control groups, bone defects were grafted with only Biogran. Guinea pigs in the control and experimental groups were serially terminated by fours on the 3 days, the 1 week, the 2 weeks, the 3 weeks, and the 4 weeks after experiment, and both sides of the mandibular bodies were removed and fixed with 10％ neutral formalin. They were decalcified and embedded in paraffin as using the usual method. The specimen sectioned and stained with hematoxylin and eosin. Also, they were radiographed with a soft X -ray apparatus. The obtained results were as follows; 1. Hemorrhagic condition, observed in the granulation tissues, disappeared on the 1 week after experiment in both groups, and more prominent in the experimental group. The granulation tissues of the experimental group had larger number of cells than those of the control group. 2. Osteoblastic differentiation in the margin of grafted material and adjacent bone was observed on the 1 week after experiment in both groups. Also, bone formation was observed in immature form on the 1 week after experiment. and more prominent in the experimental group. 3. In the polarizing microscopic examination, bone matrix was very loose on the 1 week after experiment, but increase in density with time, and more prominent in the experimental group. 4. In the microradiographic examination, newly formed bone was observed in the experimental group on the 2 weeks after experiment, and this was observed earlier than in the control group. Newly formed bone was increased with time and defected area was markedly decreased on the 4 weeks after experiment.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.89-112
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• 2002

The earliest reports of the use of electrical energy to directly stimulate bone healing seem to be in 1853 from England, the techniques involved the introduction of direct current into the non-united fracture site percutaneously via metallic needles, with subsequent healing of the defect. One endpoint of the periodontal therapy is to generate structure lost by periodontal diseases. Several procedural advances may support regeneration of attachment, however, regeneration of alveolar bone does not occur consistently. Therefore, factors which stimulate bone repair are areas for research in periodontal reconstructive therapy. Effects of cytokines or growth factors on bone repair are examples of such areas. Another one is electrical current which occurs in bone naturally, so that such bone may be particularly susceptible to electrical therapy. The purposes of this study were to observe the effects of electrical stimulation on the normal periodontium, to determine whether the electricity is the useful means for periodontal regeneration or not. Forty rats weighted about 100 gram were used and divided into 4 groups, the first group, there was no electrical stimulation with the connection of electrodes only. In the second group, there was stimulated by the 10 mA during 10 minutes per a day, in the third group was stimulated by the 25 mA , and the fourth by the 50 mA. At 3, 5, 10 and 15 days post-appliance , two rats in each group were serially sacrificed. and the maxillae and the mandible processed to paraffin, and the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows : 1. There was the distinct reversal line on the lingual alveolar crest, whereas a little changes in the labial alveolarcrest to the duration and amount of currents. 2. In 50 mA group, the cells were highly concentrated at the apex of anterior teeth, and was observed the necrotic tissue. In posterior root apex, the hypercementosis was appeared, and newly formed cementum layer has been increased continuously with the time. 3. The periodontal ligament fiber and Sharpey's fiber were arranged in order, and the bone trabeculae were increased as the experiment proceeded by, relatively the bone marrows were decreased. 4. In the pulp tissue, the blood vessels were increased with blood congestion in the experimetal specimens remarkably, and the dentinal tubules were obstructed . 5. The osteoblasts in alveolar bone proper had been showed highly activity, and also observed the formation of bone trabeculea. In the conclusion, it was suggested that the electrical stimulation has influence on the periodontium and the pulp tissue. However, there might be the injurious effects.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.1-12
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• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Journal of Periodontal and Implant Science
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• v.34 no.3
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• pp.475-487
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• 2004

Purpose: This study was aimed to evaluate the effect of the deproteinated bovine bone powder (DBBP) coated with calcium phosphate (Ca-P) on osseous regeneration in the calvarial bone defect of rat. Materials and Methods : The DBBP (Control group, n=6) and the Ca-P coated DBBP (Experimental group, n=6) were grafted in the critical sized calvarial bone defect (8 mm) of rat weighing 250 g. The animals were sacrificed at 1, 4 week. The biopsy specimens were decalcified with 5%formaldehyde and embedded in paraffin. The rats were sacrificed at 8 week received tetracycline (1 week), calcein blue (4 week), and alizarin red (7 week), and the biopsy specimens were taken. The specimens were embedded in methylmethacrylate and ground to 10 ${\mu}m$ thin sections were made. All of the specimens were stained with H & E and Masson's trichrome and examined under light microscope. The specimens at 8 week were examined under fluorescent microscope. Results : In the Control group, the grafted DBBP was surrounded with connective tissue, and osteoblasts were observed partially around the grafted particles at 1 week. At 4 week, some osteoid was observed and, new bone formation was observed at the periphery of grafted materials at 8 week, In the Experimental group, some osteoid was seen at the periphery of the grafted Ca-P coated DBBP at 1 week, and osteoblast and newly formed bone were observed around the grafted materials. At 8 week, newly formed bone was observed at the periphery of the grafted materials. Conclusion: These results suggest that Ca-P coated DBBP group was more and faster than DBBP group in new bone formation and Ca-P could contribute to enhance bone formation in the critical sized calvarial bone defect of rat.

• Journal of Periodontal and Implant Science
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• v.38 no.2
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• pp.163-170
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• 2008

Purpose: Biphasic calcium phosphates have been of great interest recently. Mixing adequate ratios of hydroxyapatite(HA) and beta-tricalcium phosphate($\beta$-TCP) allowed to control the resorption rate without distorting its osteoconductive property. This study evaluated the bone formation effect of newly developed biphasic calcium phosphate(BCP) in calvarial defect of rabbits. Materials and Methods: 6 male New Zealand rabbits were used. Four defects with 8mm in diameter were created on each animal. BCP with HA/$\beta$-TCP ratio of 7:3 and particle size of $0.5{\sim}1.0\;mm$ was used as the test group and bovine bone with $0.25{\sim}1.0\;mm$ particle size, as the control group. Both test and control group materials were randomly implanted in the calvarial defects and were covered witha polymer membrane. The animals were sacrificed after 12, 24, and 48 weeks of implantation under general euthanasia. Resin blocks were obtained and were stained by masson's trichrome for histological observation. Results: Overall results were uneventful without any defect exposure or inflammation. The amount of new bone formation and bone maturity increased with increase in healing period at both groups. New bone in test group was mostly formed along the material particle surrounded by osteoblasts, and observation of osteoblastic stream was also present. Bone maturity increased as it was closer to thedefect margins. Under the same healing period, the test group showed more bone formation than the control group with more stable bovine bone particles remaining even after 48 weeks, whereas considerable resorption took place in BCP. Almost total defect closure was observed in test group with new bone formation in the central part of the defect. However, limited new bone formation was observed in the control group. Conclusion: Within the limits of the study, the present study reveals the newly developed BCP to be a good osteoconductive material. However, further studies are needed to be conducted in a different study model with a larger sample size.

• Journal of Periodontal and Implant Science
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• v.48 no.1
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• pp.34-46
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• 2018

Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin $D_3$ on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin $D_3$ were prepared. Cytotoxic effects and osteogenic differentiation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, ALP staining, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR) for osteogenic differentiation markers such as ALP, collagen type I (Col-I), osteocalcin (OCN), vitamin D receptor (VDR), and glyceraldehyde 3-phosphate dehydrogenase. Results: The MTT assay showed that 1,25-dihydroxyvitamin $D_3$ did not inhibit cell growth and that the rate of cell proliferation was higher than in the positive control group at all concentrations. ALP activity was also higher than in the positive control group at low concentrations of 1,25-dihydroxyvitamin $D_3$ ($10^{-10}$, $10^{-12}$, and $10^{-14}M$). RT-PCR showed that the gene expression levels of ALP, Col-I, OCN, and vitamin D receptor (VDR) were higher at a low concentration of 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$) showed no significant differences in the overall degree of calcification. In contrast to the positive control group, formation of bone nodules was induced in the early stages of cell differentiation. Conclusions: We suggest that 1,25-dihydroxyvitamin $D_3$ positively affects cell differentiation and matrix mineralization. Therefore, it may function as a stimulating factor in osteoblastic bone formation and can be used as an additive in bone regeneration treatment.

• Journal of the korean academy of Pediatric Dentistry
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• v.29 no.3
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• pp.345-353
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• 2002

Bone morphogenetic proteins(BMPs) are secretory signal molecules which have a variety of regulatory functions during morphogenesis and cell differentiation. To evaluate roles of BMPs and their receptors on mouse sagittal suture development, we have examined their expression patterns in serial sections of sagittal sutures by in situ hybridization during embryonic stages(E15-E18). BMP-2 and BMP-3 were expressed in the osteogenic front and parietal bone on embryonic 15day, from E16 in hair follicle. BMP-4 was strongly expressed in the osteogenic front and weakly expressed in the mesenchyme and parietal bone. BMP-S was expressed in the hair follicles. BMP-6 was not expressed in this study. BMP-7 was expressed in parietal bone during embryonic stage. BMPR-IB was expressed in the osteogenic front, but BMPR-IA was not. From these datas, we suggest that the BMP-4 regulates the early commitment of mesenchymal cells to the osteogenic lineages, the BMP-2 and BMP-3 may be involved in regulating the differentiation of osteoblast precursor cells. BMP-7 was involved in maintenance of differentiated osteoblasts. BMPs were key signaling molecules that regulate early calvarial bone morphogenesis, mediated by BMPR-IB.

• BMB Reports
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• v.53 no.6
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• pp.329-334
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• 2020

Inflammasomes are cytosolic, multiprotein complexes that act at the frontline of the immune responses by recognizing pathogen- or danger-associated molecular patterns or abnormal host molecules. Mesenchymal stem cells (MSCs) have been reported to possess multipotency to differentiate into various cell types and immunoregulatory effects. In this study, we investigated the expression and functional regulation of NLR Family Pyrin Domain Containing 3 (NLRP3) inflammasome in human umbilical cord blood-derived MSCs (hUCB-MSCs). hUCB-MSCs expressed inflammasome components that are necessary for its complex assembly. Interestingly, NLRP3 inflammasome activation suppressed the differentiation of hUCB-MSCs into osteoblasts, which was restored when the expression of adaptor proteins for inflammasome assembly was inhibited. Moreover, the suppressive effects of MSCs on T cell responses and the macrophage activation were augmented in response to NLRP3 activation. In vivo studies using colitic mice revealed that the protective abilities of hUCB-MSCs increased after NLRP3 stimulation. In conclusion, our findings suggest that the NLRP3 inflammasome components are expressed in hUCB-MSCs and its activation can regulate the differentiation capability and the immunomodulatory effects of hUCB-MSCs.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.22 no.3
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• pp.274-287
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• 2000

The use of dental implants has increased tremendously in recent years and is expected to increase even more in the future. The successful outcome of any implant procedure is surely dependent on interrelationship of the various components of an equation that includes biocompatibility of implant material, macroscopic and microscopic nature of the implant surface, the status of implant bed, surgical technique, undisturbed healing phase and subsequent prosthetic design and long-term loading phase. The purpose of this study was to clarify the effects of adrenalectomy on the osseointegration of pure titanium implants. Seventy rats, 11 weeks of age, were divided into two groups : an adrenalectomized group and a control group. Titanium screw implant(diameter, 2.0mm; length, 3.5mm) was placed into left tibia of 70 rats, 35 in control group and 35 in the experimental group. The rats were sacrificed at different time interval (1, 2, 3, 4, 6, 8, and 12 weeks after implantation) for histopathologic observation, histomorphometric analysis and immunohistochemistry with fibronectin and CD44 antibody. The results obtained from this study were as follows: 1. Histopathogically, findings, newly formed bone was seen at 3 weeks control group and became lamellar bone at 12 weeks. At 6 weeks, lipocytes were observed in bone marrow space. Thickness of regenerated trabecular bone increased till 6 weeks after then, that decreased gradually. 2. By histomorphometric analysis, marrow bone density and contact ratio of marrow bone to implant decreased significantly from 8 to 12 weeks in experimental group compared to control group and also total bone to implant contact ratio decreased significantly from 4 to 12 weeks in experimental group compared to control group. 3. Fibronectin immunoreactivity was very strong at 3 and 4 weeks control group. And after that reduced gradually. But it was continuously strong from 1 to 12 weeks experimental group. 4. CD44 immunoreactivity was very strong in the newly formed osteoblasts at 3 and 4 weeks control group. But it reacted minimally later. However, it reacted continuously strong from 3 to 12 weeks experimental group. From these results, bone to implant contact ratio decreased gradually from 4 weeks in adrenalectomized group compared to control group. CD44 and fibronectin immunoreactivities were strong at all times in adrenalectomized rats. Therefore, it could be stated that immature bone remained continuously for a long time and not readily proceeded into mature status.