• Title/Summary/Keyword: ectopic bone formation

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Expression, Refolding, and Characterization of the Proteolytic Domain of Human Bone Morphogenetic Protein 1 (뼈형성 단백질(Bone Morphogenetic Protein 1)의 단백질 분해 부위의 발현 및 특성 연구)

  • ;Daihung Do
    • Journal of Life Science
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    • v.10 no.2
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    • pp.218-227
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    • 2000
  • Bone morphogenetic protein 1 (BMP-1) is part of a complex capable of inducing ectopic bone formation in mammals. Studies on TGF-β1 processing and Drosophila dorsal-ventral patterning have focused attention on BMP-1 as important in mediating the biological activity of this bone inducing complex. Herein, the bacterial expression, refolding, purification, and initial characterization of the BMP-1 proteolytic domain (BPD) are described. A semi-quantitative fluorescence-based thin layer chromatography assay was developed to assist in rapidly screening for optimal renaturation conditions. According to a preliminary screen for optimal conditions for the refolding of BPD , a detectable proteolytic activity against a high turnover substrate for astacin, a homologous protease from crayfish was observed. The conditions identified have allowed the expression of sufficient amounts of BPD for the characterization of the protein. Its proteolytic activity exhibits the same cleavage specificity as astacin against seven substrates that were previously synthesized for studying astacin. Furthermore, this activity is inhibited by the metal chelator 1,10-phenanthroline but not by its analogue 1,7-phenanthroline. The collagenase inhibitor Pro-Leu-Gly hydroxamate was found to inhibit both astacin and BPD activity. The results presented in this paper argue that BMP-1 does in fact possess an intrinsic proteolytic activity.

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MicroRNA-26a Regulates RANKL-Induced Osteoclast Formation

  • Kim, Kabsun;Kim, Jung Ha;Kim, Inyoung;Lee, Jongwon;Seong, Semun;Park, Yong-Wook;Kim, Nacksung
    • Molecules and Cells
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    • v.38 no.1
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    • pp.75-80
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    • 2015
  • Osteoclasts are unique cells responsible for the resorption of bone matrix. MicroRNAs (miRNAs) are involved in the regulation of a wide range of physiological processes. Here, we examined the role of miR-26a in RANKL-induced osteoclastogenesis. The expression of miR-26a was upregulated by RANKL at the late stage of osteoclastogenesis. Ectopic expression of an miR-26a mimic in osteoclast precursor cells attenuated osteoclast formation, actin-ring formation, and bone resorption by suppressing the expression of connective tissue growth factor/CCN family 2 (CTGF/CCN2), which can promote osteoclast formation via upregulation of dendritic cell-specific transmembrane protein (DC-STAMP). On the other hand, overexpression of miR-26a inhibitor enhanced RANKL-induced osteoclast formation and function as well as CTGF expression. In addition, the inhibitory effect of miR-26a on osteoclast formation and function was prevented by treatment with recombinant CTGF. Collectively, our results suggest that miR-26a modulates osteoclast formation and function through the regulation of CTGF.

AN EXPERIMENTAL STUDY ON THE BONE INDUCTION CAPACITY OF THE PORCINE BONE MATRIX-DERIVED BONE MORPHOGENETIC PROTEIN (돼지의 골기질유도 골형성단백질의 골유도능에 관한 실험적 연구)

  • Park, Young-Wook;Lee, Jong-Ho;Kim, Soo-Kyeong
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.19 no.3
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    • pp.265-286
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    • 1997
  • Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

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EXPRESSION OF CARTILAGE DERIVED MORPHOGENIC PROTEIN IN PLEOMORPHIC ADENOMA (다형성선종에서 연골유래형성단백의 발현)

  • Lee, Hwang-Jae;Kim, Chul-Hwan;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.3
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    • pp.247-255
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    • 2007
  • The bone morphogenic protein(BMP) can promote migration and growth of mesenchymal cells and initiate process for bone and cartilage formation. Cartilage-derived morphogenic protein(CDMP)-1 and -2 belong to the bone morphogenetic protein family in the transforming growth factor(TGF)-${\beta}$ superfamily. Although pleomorphic adenoma of the salivary glands is an epithelial tumor, it frequently shows ectopic cartilaginous formation with biomolecular studies. The mechanism of pathogenesis in cartilaginous formation is still controversy. We examined the expression and localization of CDMP-1 and -2, in comparison with the localization of cartilaginous matrix proteins, in human normal salivary glands and 20 cases of pleomorphic adenoma using immunohistochemical methods. The results were followed. 1. CMP-1 was immunolocalized in the striated ducts and the intercalated ducts, but not expressed in excretory duct, CDMP-2 was not expressed in the normal salivary glands. 2. CMP-1 was immunolocalized in the ductal cell and cuboidal neoplastic myoepithelial cells around the chondroid areas of the pleomorphic adenomas, whereas these molecules were not localized in the spindle-shaped neoplastic myoepithelial cells of the myxoid element in these tumors. CDMP-2 was expressed neither in normal salivary glands nor in any elements of the pleomorphic adenomas. 3. In transmission electron microscopic view, the tumor cells are composed of modifed myoepithelial cells between hyaline and myxoid stroma. 4. In Immuno-blot analysis, strong overexpression of CDMP-1 was frequently seen in pleomorphic adenomas, but the level of CDMP-2 was expressed minimally in pleomorphic adenoma. From the these results, it should be suggested that undifferentiated neoplastic myoepithelial cells around the chondroid areas expressed CDMP-1 and suggested that this molecule may play a role in the differentiation of neoplastic myoepithelial cells in pleomorphic adenoma, but not CDMP-2.

Expansile dentigerous cyst invading the entire maxillary sinus: a case report

  • Cho, Ju-Yeon;Nam, Ki-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.38 no.4
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    • pp.245-248
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    • 2012
  • Reported cases of a large dentigerous cyst involving the whole maxillary sinus are uncommon. A 22-year-old female patient suffering from swelling of the right infraorbital area and cheek with dull pain was referred to our department. Findings on computed tomography (CT) and magnetic resonance imaging (MRI) revealed a huge mass containing a displaced maxillary third molar involving the right maxillary sinus as a whole, with partial erosion of the posterior sinus cortical bone. Under general anesthesia, the mass was enucleated using the Caldwell-Luc approach, and, following histopathological analysis, was diagnosed as a dentigerous cyst. The case was followed for a period of seven years, and no evidence of sinus infection or recurring cyst formation was observed during that time.

IGF1 potentiates BMP9-induced osteogenic differentiation in mesenchymal stem cells through the enhancement of BMP/Smad signaling

  • Chen, Liang;Zou, Xiang;Zhang, Ran-Xi;Pi, Chang-Jun;Wu, Nian;Yin, Liang-Jun;Deng, Zhong-Liang
    • BMB Reports
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    • v.49 no.2
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    • pp.122-127
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    • 2016
  • Engineered bone tissue is thought to be the ideal alternative for bone grafts in the treatment of related bone diseases. BMP9 has been demonstrated as one of the most osteogenic factors, and enhancement of BMP9-induced osteogenesis will greatly accelerate the development of bone tissue engineering. Here, we investigated the effect of insulin-like growth factor 1 (IGF1) on BMP9-induced osteogenic differentiation, and unveiled a possible molecular mechanism underling this process. We found that IGF1 and BMP9 are both detectable in mesenchymal stem cells (MSCs). Exogenous expression of IGF1 potentiates BMP9-induced alkaline phosphatase (ALP), matrix mineralization, and ectopic bone formation. Similarly, IGF1 enhances BMP9-induced endochondral ossification. Mechanistically, we found that IGF1 increases BMP9-induced activation of BMP/Smad signaling in MSCs. Our findings demonstrate that IGF1 can enhance BMP9-induced osteogenic differentiation in MSCs, and that this effect may be mediated by the enhancement of the BMP/Smad signaling transduction triggered by BMP9.

ROOT MALFORMATION OF PERMANENT INCISORS BY ALVEOLAR BONE FRACTURE (치조골 골절을 동반한 유치열기 외상에 의한 영구절치의 치근 형성 이상)

  • Ji, Eun-Hye;Choi, Hyung-Jun;Choi, Byung-Jai;Son, Heung-Kyu;Kim, Seung-Hye;Song, Je-Seon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.3
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    • pp.290-295
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    • 2011
  • During tooth formation, tooth development can be affected by physical action or metabolic changes around dental follicle. Especially trauma to primary dentition is the most representative physical factor that can cause development disorders of succedaneous tooth. Enamel hypoplasia and crown discoloration of succedaneous tooth are common complications of trauma. And impaction, ectopic eruption, arrest of root formation and root dilaceration of succedaneous tooth are rare. In this case, a 6-year and 5-month-old female patient visited for dental evaluation after trauma. She was diagnosed with alveolar bone fracture near upper front teeth, extrusion of the upper right and left primary central incisors, intrusion of the upper right primary lateral incisor, and palatal luxation of the upper left primary lateral incisor. Upper right and left primary central incisors with severe mobility were extracted, with gingival suture on the day of the visit. During 24 months check up, root dilacerations were found near the cemento enamel junction in the upper lateral incisors and arrests of root formation were found on the coronal 1/3 of the root in the upper central incisors. Although alveolar bone fracture is rare type of trauma in children, a thorough examination of alveolar bone is essential for prognosis and following treatment in patients with trauma.

The Effects of rhBMP-2/ACS on the Periodontal Healing of 1-Wall Intrabony Defects in Dogs (성견 1면 골결손부에서 rhBMP-2/ACS가 치주조직 치유에 미치는 영향)

  • Jun, Hye-Young;Cho, Kyoo-Sung;Kim, Chong-Kwan;Chai, Jung-Kiu
    • Journal of Periodontal and Implant Science
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    • v.29 no.4
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    • pp.873-893
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    • 1999
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and repair of function. For more than a decade there have been many efforts to develop materials and bioactive molecule(such as growth factor and differentiation factors) to promote periodontal wound healing. Among the bioactive molecules, bone morphogenetic protein(BMP) was studied for periodontal wound healing. Since Urist demonstrated that demineralized bone matrix could induce the formation of cartilage and bone in ectopic site, many studies on BMP have been reported. Among those BMPs, it was reported that rhBMP-2 enhanced the healing of bone defects in animal studies and clinical studies. However, its efficacy in periodontal regeneration, especially 1-wall intrabony defects is still unknown. The purpose of this study was to examine the effect of rhBMP-2/ACS on the epithelial migration, gingival connective tissue adhesion, cementum formation, alveolar bone regeneration in intrabony defects of dogs. Four millimeter deep and four millimeter wide 1-wall defects were surgically created in the mesial aspects of the 3rd incisors. The test group received rhBMP-2/ACS with a flap procedure and the control underwent buffer/ACS with a flap procedure. Histologic analysis after 8 weeks of healing revealed the following results: 1. The length of epithelial growth(the distance from alveolar crest to the apical end of JE) was $0.9{\pm}1.5mm$ in the control group and $1.2{\pm}1.4mm$ in the test group. There was no statistically significant difference between the two groups. 2. The length of connective tissue adhesion was $2.4{\pm}1.3mm$ in the control group and $1.2{\pm}1.1mm$ in the test group. The control group showed significantly enhanced adhesion(P<0.05). 3. The length of new cementum was $0.9{\pm}1.0mm$ in the control group and $1.7{\pm}0.8mm$ in the test group. The test group showed significantly enhanced cementum regeneration(P<0.05). 4. The length of new bone height was $1.9{\pm}0.6mm$ in the control group and $2.4{\pm}0.9mm$ in the test group. There was no statistically significant difference between the two groups. 5. The new bone area was $4.7{\pm}1.7mm^2$ in the control group and $8.0{\pm}2.0mm^2$ in the test group. The test group showed significantly enhanced bone formed area(P<0.05). 6. The new bone density was $73.0{\pm}8.6%$ in the control group and $66.6{\pm}15.3%$ in the test group. There was no statistically significant difference between the two groups. These results suggest that the use of rhBMP-2 in 1-wall intrabony defects has significant effect on new cementum and new bone formation area, but doesn't have any significant effect on the prevention of junctional epithelium migration and new bone formation height.

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Thoracic Myelopathy and Radiculomyelopathy due to Ossification of the Ligamentum Flavum - A Cases Report - (황색인대 골화에 의한 흉추부 척수증 및 신경근척수증 - 증례보고 -)

  • Lee, Sang Dae;Rhee, Dong Youl;Jeong, Young Gyun;Park, Hyuck;Cho, Bong Soo;Kim, Soo Young
    • Journal of Korean Neurosurgical Society
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    • v.29 no.10
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    • pp.1377-1382
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    • 2000
  • Ossification of ligamentum flavum(OLF) is a type of ectopic bone formation developed within the spinal ligament. Thoracic myelopathy is common symptom of OLF but radiculopathy is uncommon. We present 3 operated cases of OLF. Two showed myelopathy and one radiculomyelopathy. Main symptoms were motor weakness, sensory disturbance, incoordination of both legs, and back pain. One patient complained of left girdle pain in T4 dermatome. Deep tendon reflex(DTR) was increased in all cases. The diagnosis was made from myelogram, CT and MRI. Sufficient posterior decompressive laminectomy and medial facetectomy were performed. Postoperative diagnosis was comfirmed by histopathological examination. One patient could return to work 3 months after operation and two patients showed satisfactory improvement 2-3 months after operation. Early diagnosis and appropriate surgical treatment were prerequisite of good outcome.

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