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Absolute Quantification of Osteogenic Genes in Rat Calvarial Defects
Christy Gliniak
Davood Varghai
Walter M Sweeney
Brendan J. Alleyne
Kate Tobin
Greg M Cooper
Arun K Gosain
University Hospitals Department of Plastic Surgery Research
2012-02-15
Presenter: Brendan Alleyne
Affidavit:
This work was done by lab personnel and students and not by residents of this department.
Director Name: Arun K Gosain
Author Category: Student
Presentation Category: Basic Science Research
Abstract Category: Craniomaxillofacial
How does this presentation meet the established conference educational objectives?
By understanding the current concepts and basic science of calvarial critical sized defects we can develop new techniques and procedures to handle topics such as craniosynostosis. This could lead to better surgical outcomes and patient care in the future.
How will your presentation be used by practicing physicians in the audience?
By further elucidating the mechanisms of healing and regeneration of critical sized defects we are provided with valuable insight into the basic science surrounding one of the most difficult areas to properly address within the field of plastic surgery. This knowledge may assist in paving the way for more appropriate therapies for craniosynostosis.
Introduction: A rat CSD model was used to test osteogenic genes in the dura mater in rats of varying age. Our goal was to identify up regulated factors in dura beneath CSDs and to identify the importance of age and time after healing.
Methods: Parietal defects (5, 6 or 8mm) were created in varied ages of neonate and adult rats. Dura was harvested at 3, 7 or 14 days after surgery. Dura was collected from beneath defects and from the intact parietal bone on contralateral sides. Absolute qPCR was performed for BMP2, FGF2, IGF1 & TGFB1 and the mRNA copy number was expressed per 100 copies of GAPDH control.
Results: In P6 rats, 3 days after surgery showed a significant increase of BMP2, FGF2 & TGF-B1 in dura under defects compared with controls. The statistically significant percent increase of factors in defect dura was 176%, 235%, and 545% for BMP2, FGF2 & TGF-B1 respectively. Gene expression in dura under defects was highest for POD 3 & 7 in P6 rats. Gene expression in defect dura was also highest in P6 rats.
Conclusions: Our results indicate increased growth factor mRNA in dura underlying defects (not IGF-1). The up regulation occurs at POD 3 and later declines. P6 rats have a higher osteogenic potential compared to older rats. Dura under defects showed changes in the ratio of factors compared to controls. This indicates that increased production of growth factors and a decrease in BMP2 and FGF2 ratios are important in regenerating bone.
