FGF-2 Stimulation Affects Calvarial Osteoblast Biology: Quantitative Analysis of Nine Genes Important for Cranial Suture Biology by Real-Time Reverse Transcription Polymerase Chain Reaction

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Abstract
Appropriately timed closure of the cranial sutures is a critical factor in normal postnatal morphogenesis of the cranial vault. Suture patency is necessary to permit rapid neonatal expansion of the cerebral hemispheres, and later ossification is important for bony protection of the cerebrum. Premature suture ossification (craniosynostosis) leads to myriad adverse functional and developmental consequences. Several murine studies have implicated dura-derived fibroblast growth factor-2 (FGF-2) paracrine signaling as a critical factor promoting physiologic posterior frontal suture fusion. In this study, the authors used real-time reverse transcription polymerase chain reaction (RT-PCR) to study an in vitro system that models the in vivo stimulation of suture calvarial osteoblasts by duraderived FGF-2. The authors advocate real-time RT-PCR as a powerful and rapid technique that offers advantages in the highly sensitive, specific, and reproducible analyses of nine genes known to be important in cranial suture biology. The genes studied were growth factors [FGF-2, transforming growth factor (TGF)-β1, TGF-β2, and TGF-β3], growth factor receptors (FGF-R1, FGF-R2, TGF-βRI, and TGF-βRII), and a marker of osteoblast differentiation (Col-IαI). These analyses provide a “snapshot” of several important genes involved in suture fusion that is more inclusive and quantitative than that which has been previously reported. (Plast. Reconstr. Surg.112: 528, 2003.)