• Title/Summary/Keyword: osteonectin

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HEALING PROCESS OF THE CALVARIAL DEFECT FILLED WITH HYDROXYLAPATITE AND TGF-β IN RAT (백서 두개골 결손부에 Hydroxylapatitie와 TGF-β 매식 후 치유과정에 관한 연구)

  • Kwon, Hyuk-Do;Lee, Dong-Kuen;Kim, Eun-Chol
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.1
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    • pp.1-12
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    • 1999
  • The purpose of this study was to evaluate the healing process of the calvarial defect filled with hydroxylapatite(HA) and $TGF-{\beta}$ in Rat. 72 Sprague-Dawly rats were divided into 3 groups, control and two experimental groups. Bony defect were artificially prepared in the calvaria of all 72 rats and followed by implantation of HA (experimental group of 24 rats) and HA+$TGF-{\beta}$(another experimental group of 24 rats) into the defects. Sequential sacrifice was performed at 1, 2, 4, 6, 8, 12 weeks of experiment. Obtained specimen was stained with Hematoxylin and Eosin, Masson's Trichrome and Immunohistochemistry. The results were as follows, 1. Granulation tissue was prominent on control group in 1 and 2 weeks. Bony defects were filled with dense fibrous tissue through the whole experimental period and osteoinduction could not be observed in all groups. 2. Inflammatory cell infiltration was prominent on control group in 1 and 2 weeks and osteoclastic activity was high in HA implanted experimental group at 1 and 2 weeks. 3. Inflammatory cell infiltration was less and maturation of fibrous tissue could be found on HA+$TGF-{\beta}$ implanted experimental group at 1 and 2 weeks. 4. Osteoconduction activity was high in HA+$TGF-{\beta}$ implanted experimental group at 2 and 4 weeks but there was no difference after 6 weeks among 3 groups. 5. In grafted site of HA+$TGF-{\beta}$ implanted group, osteonectin expression was slightly increased from 1 week to 6 weeks. In the host site, it was increased from 1 to 4weeks. 6. In grafted site of HA+$TGF-{\beta}$ implanted group, osteocalcin expression was high at 4 weeks. In the host site, we could find the difference among 3 groups. From above results, the HA with mixture of $TGF-{\beta}$ has the potentiality of promoting bone formation in the bony defect area in the rat.

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Effect of Extract of Seeds of Carthamus tinctorius L. on Mineralization in Periodontal Ligament Cells and Osteoblastic Cells (홍화씨 추출물이 치주인대세포와 조골유사세포의 골 광물화 작용에 미치는 효과)

  • Kang, Jung-Goo;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.28 no.3
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    • pp.475-489
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    • 1998
  • The main goal for the treatment of periodontal diseases is the regeneration of lost cementum, bone and connective tissue. Clinical and histological research suggests that it is possible to restore periodontal structures. Seeds of Carthamus tinctorius L. has been used for the treatment of bone fracture and osteoporosis in traditional Korean medicine. The purpose of this study is to examine the effect of extract of seeds of Carthamus tinctorius L. on mineralization in periodontal ligament cells and osteoblastic cells. Periodontal ligament cells were primarily obtained from a extracted premolars with non-periodontal diseases, Osteoblastic cells were obtained from calvariae of a fetal rat, Cells were cultured with DMEM at $37^{\circ}C$ with 5% $CO_2$ in 100% humidity incubator. Alkaline phosphatase(ALP) level and the number of calcification nodules were examined and western blot analysis using osteonectin was performed, Measurements of ALP levels and calcification nodules showed that extract of seeds of Carthamus tinctorius L. had significantly higher activity than control in all of both cells. In western blot analysis, protein expression of osteonectin indicated that extract of seeds of Carthamus tinctorius L. showed an increased pattern than control in all of both cells. From the above results, it seems that extract of seeds of Carthamus tinctorius L. has excellent effect on mineralization in periodontal ligament cells and osteoblastic cells.

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An Evaluation of Low Intensity Ultrasonic Characteristics for Arthritis Healing (저강도 초음파의 관절염 치료 적용성 평가)

  • Hong Sung-Min;Han Seung-Moo
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.29 no.6 s.237
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    • pp.787-794
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    • 2005
  • Nowadays chronic degenerative diseases such as arthritis are increasing rapidly, even though acute infectious diseases are decreasing due to the advance of modern medicine. Although many of remedies are developed for arthritis healing, there is no precise medical prescription, and pathogenesis is not examined exactly. In this study, we confirmed gene expression of BMP 1A, BMP 2B, osteonectin and MGP which are genes related with bone formation in osteoblast by using ultrasonic stimulation. Through this study, we also evaluated the fact that ultrasound could be applied to arthritis healing by making activated osteoblast induce remodeling of cartilage. As a result of this research, BMP 2B and MGP have higher rates of expression in specific ultrasound mode. In conclusion, it is expected that ultrasound could be used efficiently for healing arthritis if we use ultrasounds according to symptoms of arthritis on reasonable terms.

HISTOLOGICAL CHANGES IN THE ELONGATED BONE AFFECTED BY OSTEODISTRACTION OF THE MANDIBLE IN THE DOG (성견 하악골 절단 후 기계적 골 견인에 의해 형성된 골 신장부에 대한 시기별 조직학적 변화)

  • Baek, Sun-Ho;Ahn, Byoung-Keun;Park, Young-Ju;Park, Hee-Geon;Park, Jun-Woo;Rhee, Gun-Joo;Lee, Yong-Chan;Cho, Byoung-Ouck
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.5
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    • pp.404-416
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    • 2001
  • Purpose : Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize serially the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination. Materials and Methods : The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every $360^{\circ}$ turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. The animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned $5{\mu}m$ thick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody (Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed. Results : 1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction.

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Effects of immunosuppressants, FK506 and cyclosporin A, on the osteogenic differentiation of rat mesenchymal stem cells

  • Byun, Yu-Kyung;Kim, Kyoung-Hwa;Kim, Su-Hwan;Kim, Young-Sung;Koo, Ki-Tae;Kim, Tai-Il;Seol, Yang-Jo;Ku, Young;Rhyu, In-Chul;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.42 no.3
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    • pp.73-80
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    • 2012
  • Purpose: The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). Methods: The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7. Results: Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control. Conclusions: These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.

LOCALIZATION OF BONE MATRIX GENE mRNA IN REGENERATING BONE TISSUE DURING THE GUIDED BONE REGENERATION (골재생유도술에 의한 골재생과정에서의 골기질 유전자 발현 양상)

  • Lee, Chang-Kon;Ryoo, Hyun-Mo;Shin, Hong-In
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.3
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    • pp.240-248
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    • 1999
  • To investigate the expression pattern of noncollagenous bone matrix proteins such as osteonectin(OSN), osteopontin(OPN) and osteocalcin(OSC) mRNA during bony healing procedure induced by guided bone regeneration method, we made artificial defects on bilateral femur of rats. Then induced bony healing by application of a nonabsorbable PTFE membrane in experimental sites and without its application in control sites for 3 weeks. The mRNA expression pattern at specimens obtained at 1, 2 and 3 weeks after operation was detected by in situ hybridization method using its antisense mRNA probes. The experimental sites revealed more rapid and favorable bony healing than control sites and new bone formation was limited within defected area by inhibitory activity of bone marrow cells. In experimental sites, the OSN and OSC mRNA were expressed strongly on osteoblasts of regenerating cortical bone at 1st week and on osteoblasts lining the trabecular bone in marrow space at 3rd week, whereas, in control sites, their expression were noted on osteoblasts lining the reactively formed sponge bones at 2nd and 3rd week. In addition, the OPN mRNA was expressed on osteoblasts and osteoclasts at sites of remodeling and osteocytes of remained trabecular bone of defected area in experimental sites and on macrophages at 1st week and osteoclasts at sites of remolding at 2nd and 3rd week in control sites. The above findings suggest that the more rapid and favorable bony healing might be induced by blocking of invading fibrous connective tissue into bony defects. And the earlier expression of OSN and OSC mRNA on osteoblasts of experimental sites suggest that the formation and resorption of regenerating bone was more rapidly progressed in confined spaces made by applicate membranes.

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Characterization of Differentiation of the Supernumerary Dental Pulp Stem Cells toward the Odontoblast by Application Period of Additives (과잉치 치수유래 줄기세포의 분화제 처리 기간에 따른 상아모세포 발현 특성)

  • Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.42 no.4
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    • pp.312-318
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    • 2015
  • The aim of this study was to investigate the possibility of the supernumerary teeth for the stem cell source in dentistry. The Real Time Quantitative Reverse Transcription Polymerase Chain Reaction (Real Time qRT-PCR) method was used to evaluate the differentiation toward the odontoblast of the supernumerary dental pulp stem cells (sDPSCs). Supernumerary dental pulp stem cells were obtained from 3 children (2 males and 1 female, age 7 to 9) diagnosed that the eruption of permanent teeth was disturbed by supernumerary teeth. The common genes for odontoblasts are alkaline phosphatase (ALP), osteocalcin (OC), osteonectin (ON), dentin matrix acidic phosphoprotein 1 (DMP-1), dentin sialophosphoprotein (DSPP). The sDPSCs were treated for 0 days, 8 days and 14 days with additives and then Real Time qRT-PCR was performed in intervals of 0 days, 8 days and 14 days. The alizarin-red solution staining was performed to visualize the stained color for the degree of calcification at 7 days, 14 days, 21 days and 28 days after treating additives to the sDPSCs. From the result of the Real Time qRT-PCR, the manifestation exhibit maximum value at 8 days after additive treatment and shifted to a decrease trend at 14 days. Alizarin-red solution staining exhibit light results at 7 days after staining and generalized dark result at 14 days. Consequently, in studies with sDPSCs, appropriate treatment time of additives for Real Time qRT-PCR is 8 days. Also, a suitable period of Alizarin-red solution staining is 14 days.

Characterization of Odontoblasts in Supernumerary Tooth-derived Dental Pulp Stem Cells between Passages by Real-Time PCR (과잉치 치수유래 줄기세포의 Real-time PCR에 의한 계대간 상아질모세포 발현 특성)

  • Ji, Sangeun;Song, Sol;Lee, Joonhaeng;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.3
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    • pp.291-301
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    • 2021
  • The aim of this study is to compare the properties of odontoblast gene of early passage cells and late passage cells derived from impacted maxillary supernumerary teeth. Impacted supernumerary teeth with maxilla were extracted from 12 patients (8 males, 4 females) between 6 - 9 years old without medical history. Real-time polymerase chain reaction (PCR) was conducted to compare characterization of odontoblast cell in the 3rd and 10th passage, and between with bone inducing additive group and without additive group. Genes for odontoblasts characteristics are osteonectin (ONT), alkaline phosphatase (ALP), osteocalcin (OCN), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP). The level of gene expression was in a decreasing order of ONT, ALP, OCN, DMP-1 and DSPP in the 3rd passage, and in decreasing order of ONT, DMP-1, OCN, ALP, and DSPP in the 10th passage in the undifferentiation and differentiation group. The order of ONT, DMP-1, and OCN did not changed. ALP and DMP-1 were switched in order. ALP and DMP-1 may be used as important markers for differentiating between the 3rd passage and 10th passage cells. Considering that supernumerary tooth was extracted young age and the time required to cultured 10th passage was short, supernumerary tooth can be considered a useful donor site of dental pulp stem cells.

Gene Expression of Supernumerary Dental Pulp Related to the Subculture Speed: A Pilot Study (계대 배양 속도가 다른 과잉치 치수유래 줄기세포 간 유전자 발현 특성)

  • Lee, Yookyung;Kim, Jongsoo;Shin, Jisun;Kim, Jongbin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.2
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    • pp.219-225
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    • 2019
  • The purpose of this study was to investigate the odontoblast gene expression related to the subculture speed of supernumerary dental pulp stem cells (sDPSCs). The stem cell is undifferentiated cells which has the ability to differentiate into various cells. Specific stimulation or environment induces cell differentiation, and these differentiation leads to bone or muscle formation. 20 sDPSCs were obtained from 20 children under aseptic condition. During the culture through the 10th passage, the third passage cells which showed short subculture period and 10th passage cells which showed long subculture period were earned. Each cell was divided into differentiated group and non-differentiated group. Quantitative real-time polychain reaction (q-RT-PCR) was performed for each group. The genes related to odontoblast differentiation, Alkaline Phosphatase (ALP), Osteocalcin (OCN), Osteonectin (ONT), Dentin sialophosphoprotein (DSPP) and Dentin matrix acidic phosphoprotein 1 (DMP-1), were measured. Differentiated cells showed more gene expression levels. Undifferentiated cells showed higher gene expression level in 10th passages but differentiated cells showed higher gene expression level in 3rd passages. Cells that showed faster subculture period showed relatively lower gene expression level except for OCN and DSPP.

Simvastatin Induces Osteogenic Differentiation and Suppresses Adipogenic Differentiation in Primarily Cultured Human Adipose-Derived Stem Cells

  • Sun, So-Hyun;Lee, Il-Kyu;Lee, Jee-Won;Shim, In-Sop;Kim, Se-Hong;Kim, Kyung-Soo
    • Biomolecules & Therapeutics
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    • v.17 no.4
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    • pp.353-361
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    • 2009
  • Recent in vitro and in vivo animal studies have reported that statin, a cholesterol-lowering drug, stimulate osteogenic differentiation. In the present study, we investigated the effect of simvastatin on osteogenic and adipogenic differentiation in primarily cultured human adipose-derived stem cells (hADSCs). The simvastatin treatment significantly increased the positive cell numbers in alkaline phosphatase and von Kossa staining, and enhanced the expression levels of bone morphogenic protein (BMP)-2, core binding factor alpha 1 (cbfa1), collgen type I and osteonectin mRNAs. Lastly, hADSCs were cultured in the adipogenic media with or without simvastatin to examine the effect of simvastatin on adipogenic differentiation. In the RT-PCR analysis, there were notable decreases in mRNA expression of aP1, C/EBP-$\alpha$ and PPAR-$\gamma$ in hADSCs cultivated in simvastatin-added medium, compared to those in simvastatin-free medium. It suggests that the adipogenic differentiation was significantly inhibited by simvastatin treatment. These observations indicate that simvastatin induces osteogenic differentiation and suppresses adipogenic differentiation in hADSCs.