• Title/Summary/Keyword: rat calvarial defect model

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Expression of transforming growth factor-1 in bone regeneration after the implantation of particulate dentin and plaster of Paris

  • Huh, Young-Chul;Kim, Su-Gwan;Kim, Jeong-Sun;Yoon, Jung-Hoon;Kim, Do-Kyung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.1
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    • pp.27-35
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    • 2006
  • Purpose: This study was performed to investigate the expression of the transforming growth factor (TGF)-1, in a rat calvarium defect model using particulate dentin and/or plaster of Paris, and correlate the bone regeneration process with the histologic events. Materials and Methods: Thirty-two Sprague-Dawley rats were divided into 4 groups of 8 animals each. A 1.0 cm-sized calvarial defects were made and the defect was filled with different graft materials as follows : Group A, the defects were filled with a mixture of particulate dentin and plaster of Paris with a 2:1 ratio; Group B, the defects were filled with plaster of Paris only; Group C, defects were filled with particulate dentin only; Group D, untreated control group. The animals were sacrificed by 1, 2, 4, 8 weeks after implantation. Excised wound tissues were processed for histology, immunohistochemistry and RT-PCR for the analysis of TGF-1 expression. Results: Gene expression of TGF-1 was detected for all experimental groups. The highest gene expression was observed in the specimen taken at the first week after implantation in Group A. According to the histologic and immunohistochemical studies, TGF-1 positive osteoblast-like cells were found in the early stage of healing after the implantation of particulate dentin and plaster of Paris. Conclusion: These findings suggest that TGF-1 may be related to new bone formation at the early healing process after the implantation of particulate dentin and plaster of Paris.

The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model (백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향)

  • Nam, Jeong-Hun;Noh, Kyung-Lok;Pang, Eun-O;Yu, Woo-Geun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Seong-Gon;Park, Young-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.366-374
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    • 2010
  • Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

Evaluation the Effectiveness of Fibrinogen to Overcome Bone Radiation Damage (방사성골괴사 극복을 위한 피브린지지체의 효용성 평가)

  • Jung, Hong-Moon
    • Journal of the Korean Society of Radiology
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    • v.15 no.4
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    • pp.539-545
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    • 2021
  • Radiation therapy is accompanied by adverse radiation effective. In particular, it is accompanied by disorders of the vascular system. Therefore, oxygen and nutrient deficiency occurs in the regeneration area. Eventually, osteoradionecrosis is formed in this cellular environment. According to a precedent study, bone morphogenetic protein-2 is used to overcome osteoradionecrosis. The purpose of this study was to investigate the regeneration ability of osteoradionecrosis by treating bone-forming protein-2 on a fibrinogen scaffold which is a biomaterial that is frequently used for bone regeneration after irradiation of the rat head. In addition, the purpose of this study was to verify the bone regeneration effect from the eight weeks. According to the experimental results, in the calvarial defected model of the irradiated mouse, making bone-formation was obtained after 8 weeks rather than bone-formation period in the early 4 weeks. moreover, it was found that the regenerated bone formation of the fibrinogen scaffold is formed from the inside of the bone of the defect area.

Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 Targeted-microRNA-221

  • Chang, Woochul;Kim, Ran;Park, Sang In;Jung, Yu Jin;Ham, Onju;Lee, Jihyun;Kim, Ji Hyeong;Oh, Sekyung;Lee, Min Young;Kim, Jongmin;Park, Moon-Seo;Chung, Yong-An;Hwang, Ki-Chul;Maeng, Lee-So
    • Molecules and Cells
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    • v.38 no.7
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    • pp.643-650
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    • 2015
  • The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule-1 was upregulated by microRNA-221 in mesenchymal stem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targeted-intercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells

  • Katagiri, Wataru;Kawai, Takamasa;Osugi, Masashi;Sugimura-Wakayama, Yukiko;Sakaguchi, Kohei;Kojima, Taku;Kobayashi, Tadaharu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.39
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    • pp.8.1-8.8
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    • 2017
  • Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.