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| Status |
Public on Jun 30, 2011 |
| Title |
Gene Expression Profiles of Jaw and Alveolar Bones Are Uniquely Different from Those of Trunk and Limb Bones |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Certain bone graft materials are employed for alveolar bone regeneration, and autografts are commonly used. Alveolar and jaw bones are developmentally derived from neural crest cells, while most trunk and limb bones are derived from mesodermal mesenchymal cells. Consequently, the chosen bone graft material is likely to have a different developmental origin than the recipient bone. We hypothesized that there are differences in the gene expression profiles and bone healing capacities between bones with different developmental origins. Therefore, we investigated these properties in the maxilla, mandible, ilium and femur. DNA microarray data revealed close homology between the gene expression profiles of the ilium and femur. The gene expressions of Wnt-1, P75, SOX10, Nestin and Musashi-1 were significantly higher in the maxilla and mandible than in the ilium and femur. The frontal bone healed faster than the parietal bone. Parietal bone defects transplanted with maxillary and mandibular bone grafts exhibited closure. Thus, it is suggested that the maxilla and mandible have different gene expression profiles from the other bones examined, and exhibit neural crest cell properties with a marked healing ability in adults. The data further suggest that jaw bones are effective as both bone graft materials and graft beds.
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| Overall design |
Animals:
Twenty-five 5-week-old male C57BL/6J mice were randomly divided into five groups (n=5 each) based on the bone graft materials as follows: group 1, ilium group; group 2, femur group; group 3, maxilla group; group 4, mandible group; group 5, negative control group with no implanted material. All the mice were sacrificed after 4 weeks. All the animal procedures were approved by the Animal Research Committee of Showa University, Tokyo, Japan.
RNA Isolation:
Bones were extirpated from 5-week-old mice and the regenerated tissue in the bone graft areas were extirpated from 9-week-old mice that were grafted bones into bone defects. Total RNA was isolated from the bones and tissue using an RNeasy Lipid Tissue Midi Kit (Qiagen, Hilden, Germany) .
DNA Microarray:
A Whole Mouse Genome Oligo Microarray (Agilent Technologies Inc., Santa Clara, CA, USA), which contains more than 41,252 probe sets, was used to probe the comprehensive gene expression profiles of the maxilla, mandible, ilium and femur. The expression analysis file created from each sample was imported into GeneSpring 10.0.2 (Agilent Technologies Inc.).
Quantitative Real-Time PCR:
We performed quantitative real-time PCR of the maxilla, mandible, ilium and femur, as well as the regenerated tissue in the bone graft areas. An Omniscript Reverse Transcriptase Kit (Qiagen, Valencia, CA, USA) was used to create cDNAs. Real-time PCR was performed for 50 cycles of 95°C for 15 s and 60°C for 1 min, followed by 50°C for 2 min and 95°C for 10 min. We used primers for six genes (Wnt-1, P75, SOX10, Nestin, Musashi-1 and GAPDH). Wnt-1, P75 and SOX10 are neural crest cell markers (Stemple and Anderson, 1992; Kelsh, 2006; Yoshida et al., 2008), while Nestin and Musashi-1 are nerve stem cell markers (Sakakibara et al., 1996; Kawaguchi et al., 2001). The gene expression levels of these five genes were determined after normalization by the expression levels of the control gene GAPDH.
Graft Material Procedure:
All surgical procedures were performed under general anesthesia in sterile conditions. After inhalation of ethyl ether, general anesthesia was achieved with an intraperitoneal injection of pentobarbital sodium. Bones for the graft materials were removed and granulated. The amounts of graft materials transplanted in the four groups were approximately 0.2 g.
Surgical Procedure:
An incision was made in the head, and a full-thickness flap was created exposing the calvarial bone. A diamond bur was used to perforate an area of 2 mm in diameter on the mouse frontal bone and parietal bone, avoiding perforation of the dura mater (Figure 3A). Subsequently, one of the four materials was grafted into each frontal and parietal bone defect. The negative control group did not receive implantation of a graft material (Figure 3B). All of the graft materials were transplanted into the defects within 30 min after extirpation. The flap was repositioned and sutured tightly with non-absorbable sutures, covering the bone defect.
Micro-computed Tomography (µCT) Observations:
A µCT system (eXplore Locus CT System and MicroView; GE Healthcare, Tokyo, Japan) was used to observe new bone formation. Images were acquired immediately after surgery and after 4 weeks.
Histologic Procedures:
The animals were euthanized at 4 weeks after surgery with an overdose of ethyl ether. All of the defects in the groups were dissected together with the surrounding soft and hard tissues. Section blocks were fixed with 4% paraformaldehyde, decalcified for 2 days, neutralized with 5% anhydrous sodium sulfate and then embedded in paraffin. Sections were cut, deparaffinized and stained with hematoxylin and eosin.
Immunohistochemical Procedures:
Sections were cut, mounted on slides, deparaffinized and incubated with an anti-Wnt-1 antibody (AnaSpec Inc., San Jose, CA, USA) at appropriate dilutions.
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| Contributor(s) |
Taguchi T, Watahiki J, Nampo T, Yamamoto G, Enomoto A, Ono M, Irie T, Tachikawa T, Maki K |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Jun 22, 2011 |
| Last update date |
Jan 12, 2017 |
| Contact name |
Tomohiro Taguchi |
| Organization name |
Showa University
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| Department |
School of Dentistry
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| Street address |
1-5-8 Hatanodai
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| City |
Shinagawa-ku |
| State/province |
Tokyo |
| ZIP/Postal code |
142-8555 |
| Country |
Japan |
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| Platforms (1) |
| GPL7202 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Probe Name version) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA143907 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE30167_RAW.tar |
31.7 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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