This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.
This study was designed to evaluate the bone formation capability of the bone substitute when compared with autogenic bone, freeze-dried demineralized allogeneic bone and bioglass into parietal bone of the rats. We made the parietal bone defects in $7{\times}7mm$ size on rats and has performed the bone graft in each experimental groups. Postoperatively 1, 2, 4, 6, 8, weeks, each specimen stained with H & E, Masson's trichrome methods. We evaluated the osteogensis capability in each groups. The result were as follow : 1. Inflammatory cell infiltration approached at 1 week and disappeared at 4 weeks in all experimental group, expecially severe in freeze-dried demineralized allogeneic bone group. 2. New capillry proliferation was increased in autogeneic bone graft group than any other groups and was increased till 2 weeks and decreased in freeze-dried demineralized allogeneic bone group and was few in bioglass group. 3. Osteoblastic activity increased in autogeneic bone and freeze-dried demineralized allogeneic bone groups till 4 weeks, and decreased in 6 weeks which no difference between these groups. But, few occurred in bioglass group till 6 weeks. 4. Initial osteoclastic activity was prominent in freeze-dried demineralized allogeneic bone group and few in autogeneic bone group. 5. New bone formation bega at 1 week in autograft and freeze-dried demineralized allogenic bone groups, but, mild new bone formation at 8 weeks in bioglass.
Osteoblasts are specialized mesenchymal cells that are responsible for bone formation. In this study, we examine the role of GATA4 in osteoblast differentiation. GATA4 was abundantly expressed in preosteoblast cells and gradually down-regulated during osteoblast differentiation. Overexpression of GATA4 in osteoblastic cells inhibited alkaline phosphatase activity and nodule formation in osteogenic conditioned cell culture system. In addition, overexpression of GATA4 attenuated expression of osteogenic marker genes, including Runx2, alkaline phosphatase, bone sialoprotein, and osteocalcin, all of which are important for osteoblast differentiation and function. Overexpression of GATA4 attenuated Runx2 promoter activity, whereas silencing of GATA4 increased Runx2 induction. We found that GATA4 interacted with Dlx5 and subsequently decreased Dlx5 binding activity to Runx2 promoter region. Our data suggest that GATA4 acts as a negative regulator in osteoblast differentiation by downregulation of Runx2.
Jeong, Hye-In;Lee, Mi Jee;Nam, Woong;Cha, In-Ho;Kim, Hyung Jun
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.43
no.5
/
pp.312-317
/
2017
Objectives: In order to assess clinical behavior, response to treatment, and factors influencing prognosis of Korean patients with osteosarcoma of the jaws (OSJ). Materials and Methods: A retrospective study of clinical, and pathological records of 26 patients with OSJ treated at the Department of Oral and Maxillofacial Surgery in Yonsei University Dental Hospital from 1990 to March 2017. Results: Of 26 patients, there were 9 men (34.6%) and 17 women (65.4%). Twenty-one of 26 patients had osteosarcoma of the mandible, and 5 of 26 patients had osteosarcoma of the maxilla. The histopathology of OSJ is highly variable, ranging from chondroblastic type (6 out of 26), osteoblastic type (10 out of 26), fibroblastic type (2 out of 26), to the rare variants like mixed type, small cell osteosarcoma types and more. All patients underwent gross total excision and only a few patients underwent neoadjuvant chemotherapy. Postoperative chemotherapy was given to most of the patients as adjuvant treatment or in combination with radiotherapy. The overall survival rate was 73.1% with an overall 2-year survival rate of 83.3%. The overall 5-,10-,15-year survival rates in this study were 73.5%, 73.5%, 49%, respectively. Using Kaplan-Meier analysis with log rank tests, the size of tumor (T-stage), and resection margins were found to affect the survival rate significantly. The chemotherapy was not significantly associated with improved survival rate. Conclusion: Surgical resection with a clear margin is the most important factor in disease survival. The role of chemotherapy and radiotherapy in OSJ remains controversial, and deserves further studies.
Park, Seung-Won;Goo, Tae-Won;Choi, Gwang-Ho;Kang, Seok-Woo;Kim, Sung-Wan;Kim, Seong-Ryul
International Journal of Industrial Entomology and Biomaterials
/
v.27
no.1
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pp.159-165
/
2013
Bone morphogenetic proteins (BMPs) belong to the transforming growth factor (TGF-${\beta}$) superfamily and are involved in osteoblastic differentiation. The largest TGF-${\beta}$ superfamily subgroup shares genetic homology with human BMPs (hBMPs) and silkworm decapentaplegic (dpp). In addition, hBMPs are functionally interchangeable with Drosophila dpp. Bombyx mori dpp may induce bone formation in mammalian cells. To test this hypothesis, we synthesized the 1,285-base pairs cDNA of full-length B. mori dpp using total RNAs obtained from the fat body of 3-day-old of the $5^{th}$ instar larvae and cloned the cDNA into the pCEP4 mammalian expression vector. Next, B. mori dpp was expressed in C3H10T1/2 cells. The target cells transfected with the pCEP4-Bm dpp plasmid showed biological functions similar to those of osteogenic differentiation induction growth factors such as hBMPs. We determined the relative mRNA expression rates of Runt-related transcription factor 2 (RUNX2), osterix, osteocalcin, and alkaline phosphatase (ALP) to validate the osteoblast-specific differentiation effects of B. mori dpp by performing quantitative real-time RT-PCR. Interestingly, mRNA expression levels of the 3 marker genes except RUNX2, in cells expressing B. mori dpp were much higher than those in control cells and C3H10T1/2 cells transfected with pCEP4. These results suggested that B. mori dpp signaling regulates osterix expression during osteogenic differentiation via RUNX2-independent mechanisms.
It is the aim of this study to investigate the effects of sodium fluoride and sodium orthovanadate upon the proliferation and activity of the osteoblast (MC3T3-E1 cells). MC3T3-E1 cells were cultured in $\alpha-MEM$ containing $10\%$ FBS and various concentration of sodium fluoride and sodium orthovanadate was appended to serum free media. DNA synthesis was examined through the $[^3H]$ thymidine incorporation into DNA. Collagen synthesis was examined through the $[^3H]$ proline incorporation into collagenase digestible protein and noncollagen protein. The following results were drawn; 1. Sodium fluoride stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M$ to $10{\mu}M$ (P < 0.005). 2. Sodium orthovanadate stimulated the DNA synthesis of osteoblast significantly in dose-dependent manner within the concentration from $2{\mu}M\;to\;8{\mu}M$, however showed diminution at $10{\mu}M$ (P < 0.001). 3. Sodium fluoride and sodium orthovanadate stimulated the percent collagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M$ to $10{\mu}M$ (P < 0.001). 4. Sodium fluoride and sodium orthovanadate stimulated the noncollagen synthesis of osteoblast significantly in dose-dependent manner within the concentration from $5{\mu}M\;to\;10{\mu}M$ (P < 0.001). In conclusion, sodium fluoride and sodium orthovanadate stimulate the proliferation and activity of osteoblast by stimulation of DNA synthesis and collagen and noncollagen synthesis in osteoblast.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.39
no.2
/
pp.55-62
/
2013
Bone tissue engineering is one of the important therapeutic approaches to the regeneration of bones in the entire field of regeneration medicine. Mesenchymal stem cells (MSCs) are actively discussed as material for bone tissue engineering due to their ability to differentiate into autologous bone. MSCs are able to differentiate into different lineages: osteo/odontogenic, adipogenic, and neurogenic. The tissue of origin for MSCs defines them as bone marrow-derived stem cells, adipose tissue-derived stem cells, and, among many others, dental stem cells. According to the tissue of origin, DSCs are further stratified into dental pulp stem cells, periodontal ligament stem cells, stem cells from apical papilla, stem cells from human exfoliated deciduous teeth, dental follicle precursor cells, and dental papilla cells. There are numerous in vitro/in vivo reports suggesting successful mineralization potential or osteo/odontogenic ability of MSCs. Still, there is further need for the optimization of MSCs-based tissue engineering methods, and the introduction of genes related to osteo/odontogenic differentiation into MSCs might aid in the process. In this review, articles that reported enhanced osteo/odontogenic differentiation with gene introduction into MSCs will be discussed to provide a background for successful bone tissue engineering using MSCs with artificially introduced genes.
Multiple bone metastases are common manifestation of many malignant tumors such as lung cancer, breast cancer, prostate cancer and renal cell carcinoma. Bone metastasis is secondary cancer in the bone, and it can lead to bone pain, fracture, and instability of the weight bearing bones, all of which may profoundly reduce physical activity and life quality. Treatment for bone metastasis is determined by multiple factors including pathology, performance status, involved site, and neurologic status. Treatment strategies for bone metastasis are analgesics, surgery, chemotherapy and radiotherapy. External beam radiotherapy has traditionally been an effective palliative treatment for localized painful bone metastasis. However, in some cases such as multiple bone metastases, especially osteoblastic bone metastasis originated from breast or prostate cancer, the radiopharmaceutical therapy using $^{89}Sr$, $^{186}Re$, $^{188}Re$, $^{153}Sm$ and $^{117m}Sn$ are also useful treatment option because of administrative simplicity (injection), few side effects, low risk of radiation exposure and high response rate. This article offers a concise explanation of the radiopharmaceutical therapy for multiple bone metastases.
Jo, Seon-Ae;Gang, Seong-Min;Park, Su-A;Lee, Hae-Shin
Proceedings of the Korean Vacuum Society Conference
/
2011.08a
/
pp.202-202
/
2011
The research of 3-dimensional (3-D) scaffold for tissue engineering has been widely investigated as the importance of the 3-D scaffold increased. 3-D scaffold is needed to support for cells to proliferate and maintain their biological functions. Furthermore, its architecture defines the shape of the new bone and cartilage growth. Polycaprolactone (PCL) has been one of the most promising materials for fabricating 3-D scaffold owing to its excellent mechanical property and biocompatibility. However, there are practical problems for using it, in vitro and in vivo; extracellular matrix components and nutrients cannot penetrate into the inner space of scaffold, due to its hydrophobic property, and thus cell seeding and attachment onto the inner surface remain as a challenge. Thus, the surface modification strategy of 3-D PCL scaffold is prerequisite for successful tissue engineering. Herein, we utilized a mussel-inspired approach for surface modification of 3-D PCL scaffold. Modification of 3-D PCL scaffolds was carried out by simple immersion of scaffolds into the dopamine solution and stimulated body fluid, and as a result, hydroxyapatite-immobilized 3-D PCL scaffolds were obtained. After surface modification, the wettability of 3-D PCL scaffold was considerably changed, and infiltration of the pre-osteoblastic cells into the 3-D scaffold followed by the attachment onto the surface was successfully achieved.
Hayashi, Y.;Yamada, S.;Ohara, N.;Kim, S-K.;Ikeda, T.;Yanagiguchi, K.;Matsunaga, T.
Proceedings of the KACD Conference
/
2003.11a
/
pp.545-545
/
2003
Chitosan is applied as a dressing for oral mucous wound and a tampon following radical treatment of maxillary sinus. Furthermore, it is being investigated as an absorbing membrane for endodontic and periodontic surgeries. A few studies have reported osteoconduction and osteogenesia at the site of chitosan implant in vivo. However, compared with soft tissue healing processes, the mechanisms concerning effects of chitosan for biological mineralization have not yet been resoil In the present study, we studied the gene expression pattern using cDNA microarray and RT-PCR analyses in hard tissue forming osteoblasts cultured with water-soluble and low molecular weight chitooligosaccharide. cDNA microarray analysis revealed that 16 genes were expressed at 〉1.5-fold higher signal ratio levels in the experimental group compared with the control group after 3 days. RT-PCR analysis showed that chitosan oligomer induced an increase in the expression of two genes, CD56 antigen and tissue-type plasminogen activator. Furthermore, the expression of mRNAs for BMP-2 was almost identical in the experimental and control groups after 3 days of culture, but slightly increased after 7 days of culture with chitosan oligomer. These results suggest that a super-low concentration of chitooligosaccharide could modulate the activity of osteoblastic cells through mRNA levels and that the genes concerning cell proliferation and differentiation can be controlled by water-soluble chitosan.
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