• The Journal of Korean Academy of Prosthodontics
• /
• v.46 no.6
• /
• pp.620-627
• /
• 2008

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

• Journal of Periodontal and Implant Science
• /
• v.50 no.2
• /
• pp.106-120
• /
• 2020

Purpose: A new form of porous polyethylene, characterized by higher porosity and pore interconnectivity, was developed for use as a tissue-integrated implant. This study evaluated the effectiveness of porous polyethylene blocks used as an onlay bone graft in rabbit mandible in terms of tissue reaction, bone ingrowth, fibrovascularization, and graft-bone interfacial integrity. Methods: Twelve New Zealand white rabbits were randomized into 3 treatment groups according to the study period (4, 12, or 24 weeks). Cylindrical specimens measuring 5 mm in diameter and 4.5 mm in thickness were placed directly on the body of the mandible without bone bed decortication, fixed in place with a titanium screw, and covered with a collagen membrane. Histologic and histomorphometric analyses were done using hematoxylin and eosin-stained bone slices. Interfacial shear strength was tested to quantify graft-bone interfacial integrity. Results: The porous polyethylene graft was observed to integrate with the mandibular bone and exhibited tissue-bridge connections. At all postoperative time points, it was noted that the host tissues had grown deep into the pores of the porous polyethylene in the direction from the interface to the center of the graft. Both fibrovascular tissue and bone were found within the pores, but most bone ingrowth was observed at the graft-mandibular bone interface. Bone ingrowth depth and interfacial shear strength were in the range of 2.76-3.89 mm and 1.11-1.43 MPa, respectively. No significant differences among post-implantation time points were found for tissue ingrowth percentage and interfacial shear strength (P>0.05). Conclusions: Within the limits of the study, the present study revealed that the new porous polyethylene did not provoke any adverse systemic reactions. The material promoted fibrovascularization and displayed osteoconductive and osteogenic properties within and outside the contact interface. Stable interfacial integration between the graft and bone also took place.

• Molecules and Cells
• /
• v.40 no.6
• /
• pp.386-392
• /
• 2017

Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

• Archives of Plastic Surgery
• /
• v.32 no.3
• /
• pp.335-342
• /
• 2005

This study was undertaken to investigate possibility of the allogenic type I collagen inducing osteoinduction or osteoconduction at critical sized bone defect in the rabbit. Twenty Newzealand white rabbit, weighted from 2.8 kg to 3.5 kg, were used in this study. The skull was exposed and two bony defects were created with diameter of 10 mm. Group I(n=10), the bony defects was grafted from the other side bone. Group II(n=10), the bony defects was grafted by the allogenic type I collagen with bone morphogenic protein(BMP). Group III(n=10), the bony defects was grafted by the allogenic type I collagen only. Group IV(n=10), the bony defects was lefted with no grafts. The grafted bones and allogenic type I collagen were investigated with radiologic densitometry, histologic analysis and immunohistochemistry after 12 weeks. No major difference was observed in the gross finding between Group I, II, III, but dura mater was exposed in bony defect,the Group IV. The radiologic study demonstrated more bony opacity in the Group I, but the other groups did not demonstrate a significant difference. In the histologic study, grafted bone edge was completely consolidated with original bone in group I and new bone ingrew into the grafted allogenic type I collagen(group II, III),but there is no bone regeneration from the original bony edge in the group IV. The percent of the new bone formation by cross-sectional area was considered statistically significant at a p value of less than 0.05(p<0.05). In the immunohistochemistry study about BMP antibodies, the group IV demonstrated osteogenic activity in front of advancing original bone edge, in which the osteoblast stained strongly for BMP antibodies, but other group does not demonstrated any osteoblastic expression. There was no immunologic rejection. In conclusion, this results do not demonstrate that the allogenic type I collagen is useful for bone substitute, but the characters of the collagen, such as pliability, easy-handling, sponge-like structure, are useful in interpositional bone graft substitutes. The further evaluation of long term results about the resorption, immunologic tissue reaction, response of applied tissue growth factor to the allogenic collagen is needed.

• Applied Microscopy
• /
• v.31 no.3
• /
• pp.287-305
• /
• 2001

To investigate a role of cartilage canals in osteogenesis and growth of the vertebrae, in human fetuses ranging from 50 mm to 260 mm crown rump length were studied by electron microscopy. The initial appearance of cartilage canals of the vertebral body was observed at 60 mm fetus. In 80 mm fetus, primary ossification center in the vertebral body was first noted. The vertebral body showed calcified chondrocytes surrounded by a tone of hypertrophied chondrocytes and deep canals which terminated in calcified matrix. Most hypertrophied chondrocytes in the centrum showed in various stage of degeneration in disorderly arrangement. At the blind end of deep canal, osteogenic cells, osteoblasts and chondroclasts were observed. Resorption of unmineralized cartilage septa was undertaken by perivascular cells within cartilage canals. The ruffled border of the chondroclast was restricted to resorption site of calcified cartilagenous matrix. The periosteal bone formation was followed by the appearance of primary center of the centrum at 120 mm fetus. The osteoblasts of the perichondrium started to lay down a thin membranous bony lamella on the outer surface of the osseous trabeculae of the centrum. The processes of bone formation in the vertebral bodies were found to possess morphological similarities to that occurring at secondary center of the epiphysis of a long bone. These results indicate that the connective tissue cells within the cartilage canals proliferate and differentiate into osteoblasts at the site of endochondral ossification of the vertebrae.

• Journal of Periodontal and Implant Science
• /
• v.23 no.2
• /
• pp.315-330
• /
• 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
• /
• v.27 no.2
• /
• pp.363-377
• /
• 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.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.27 no.2
• /
• pp.300-308
• /
• 2000

The Fibroblast growth factors(FGFs) plays an important role in the control of osteogenesis during skeletal development. Especially, FGF-2 is a potent mesodermal inducer during embryogenesis and FGF receptors (FGFRs) messages are strongly expressed in developing bones. In this study, we investigated the effect of bFGF on osteopontin(OPN) gene expression in ST-2 cells and tried to elucidate the mechanism of its stimulatory effects. The obtain results were as follows; The treatment of bFGF(1ng/ml) upregulates OPN, fibronectin mRNA levels and downregulates type I collagen mRNA levels. But, there was no remarkable difference in alkaline phosphatase mRNA levels between two groups. The OPN gene expression increased in a dose-dependent manner up to 10ng/ml and OPN gene began to occur at around 3h with continuous increase up to 24h then decreased to basal level at 48h. 30 minutues pretreatment with cycloheximide (500ng/ml), a protein synthesis inhibitor, prior to addition bFGF resulted in blocking bFGF induced OPN expression. These results suggest that bFGF increased the level of OPN mRNA in a dose and time-dependent manner via the synthesis of certain transcriptional regulatory proteins.

• The korean journal of orthodontics
• /
• v.26 no.3
• /
• pp.291-299
• /
• 1996

Orthodontic force is a mechanical stress controlling both of tooth movement and skeletal growth. The mechanical stress stimulate bone cells that may exert some influence on bone remodeling. The purpose of this study was to evaluate the difference in cellular activity depending on mechanical stresses such as compressive and tensile force by determining the alkaline phosphatase(ALP) activity. A clonal osteogenic cell line MC3T3-E1 was seeded into a 24-well plate($2{\times}10^4/well$). At the confluent phase, a continuous compressive hydrostatic pressure($25g/cm^2$, $300g/cm^2$) and continuous tensile hydrostatic pressure($-25g/cm^2$, $-300g/cm^2$) were applied for 4, 6, 10, 14, 18, 20 days respectively by a diaphgragm pump. At the end of the stimulation period, cell layers were prepared for ALP activity assay. The ALP activity of the compressive group increased more than that of the tensile group at same force magnitude, whereas the cells responded to a similar pattern regardless of the type of mechanical stress The ALP activity of the compressive and tensile group turned into the level of the control group as the length of time increased. These results indicated that a mechanical stress may be more effective on cellular activity during active cellular proliferation and differentiation periods. The time to achieve maximum ALP activity was delayed as the mechanical stress increased in both the compressive and the tensile group. Accordingly, the magnitude of the stress rather than the type of mechanical stress may have more influence on cellular activity.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.34 no.2
• /
• pp.220-229
• /
• 2008

Purpose: The present study was aimed to examine the effect of acellular dermal matrix ($AlloDerm^{(R)}$) grafted to the experimental tissue defect on tissue regeneration. Materials and Methods: Male albino rabbits were used. Soft tissue defects were prepared in the external abdominal oblique muscle. The animals were then divided into 3 groups by the graft material used: no graft, autogenous dermis graft, and $AlloDerm^{(R)}$ graft. The healing sites were histologically examined at weeks 4 and 8 after the graft. In another series, critical sized defects with 8-mm diameter were prepared in the right and left iliac bones. The animals were then divided into 5 groups: no graft, grafted with autogenous iliac bone, $AlloDerm^{(R)}$ graft, $AlloDerm^{(R)}$ graft impregnated with rhBMP-2, and $AlloDerm^{(R)}$ graft with rhTGF-${\beta}1$. The healing sites of bone defect were investigated with radiologic densitometry and histological evaluation at weeks 4 and 8 after the graft. Results: In the soft tissue defect, normal healing was seen in the group of no graft. Inflammatory cells and foreign body reactions were observed in the group of autogenous dermis graft, and the migration of fibroblasts and the formation of vessels into the collagen fibers were observed in the group of $AlloDerm^{(R)}$ graft. In the bone defect, the site of bone defect was healed by fibrous tissues in the group of no graft. The marked radiopacity and good regeneration were seen in the group of autogenous bone graft. There remained the traces of $AlloDerm^{(R)}$ with no satisfactory results in the group of $AlloDerm^{(R)}$ graft. In the groups of the $AlloDerm^{(R)}$ graft with rhBMP-2 or rhTGF-${\beta}1$, there were numerous osteoblasts in the boundary of the adjacent bone which was closely approximated to the $AlloDerm^{(R)}$ with regeneration features. However, the fibrous capsule also remained as in the group of $AlloDerm^{(R)}$ graft, which separated the $AlloDerm^{(R)}$ and the adjacent bone. Conclusions: These results suggest that $AlloDerm^{(R)}$ can be useful to substitute the autogenous dermis in the soft tissue defect. However, it may not be useful as a bone graft material or a carrier, since the bone defect was not completely healed by the bony tissue, regardless of the presence of osteogenic factors like rhBMP-2 or rhTGF-${\beta}1$.