title: Iron-induced rat coronal suture fusion in vitro: the role of redox regulation.
abstract: The presumptive coronal sutures of rat fetuses at gestation days 19 and 20 have been shown to fuse prematurely when grown in the absence of dura mater in culture. In the present study, the representative enzymes of glucose metabolism and the antioxidative pathway were assayed during the process of suture fusion. The coronal sutures of fetal day 19.5 (F19) and neonatal day 1 rats were grown in the presence or absence of dura mater in serum-free culture. The enzymes assayed were hexokinase (HK) and pyruvate kinase (PK) of glycolysis, and glucose 6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) of the antioxidative pathway. F19 sutures cultured without dura mater, which fused, showed significant increases in enzyme activities over the preculture levels. HK increased by 200% to 300% of the preculture levels, G6PD by 400% to 500%, GR by 200%, and PK by 400% to 500%. The fetal sutures cultured with dura mater, which did not fuse, showed little alterations of HK, G6PD, and GR activities, but showed a significant 200% to 400% increase in PK activity. Neonatal sutures showed significant increases in enzyme activities during culture, but the presence of dura mater did not significantly affect enzyme activities. High activity levels of enzymes of the antioxidative pathway in F19 sutures coincided with the period of premature suture fusion. Treatment of fetal calvaria with prooxidant (induced by ferrous iron and ascorbic acid) produced suture fusion even in the presence of dura mater. Treatment with deferoxamine (an iron chelator and antioxidant) during the culture prevented suture fusion. The results suggest that fusing sutures experience increased biosynthetic demands and are placed under oxidative stress. When oxidative stress overwhelms the dural influence, the sutures undergo premature fusion.
authors.affiliation: Division of Plastic, Reconstructive and Maxillofacial Surgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: M J
authors.initials: MJ
authors.lastname: Im
authors.forename: J M
authors.initials: JM
authors.lastname: Winograd
authors.forename: P N
authors.initials: PN
authors.lastname: Manson
authors.forename: C A
authors.initials: CA
authors.lastname: Vander Kolk
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001205:Ascorbic Acid / Q000494:pharmacology; D003393:Cranial Sutures / Q000187:drug effects* / Q000196:embryology / Q000201:enzymology*; D003398:Craniosynostoses / Q000196:embryology / Q000201:enzymology* / Q000209:etiology*; D003470:Culture Media; D046508:Culture Techniques; D004388:Dura Mater / Q000502:physiology; D005260:Female; D005296:Ferrous Compounds / Q000494:pharmacology; D007501:Iron / Q000494:pharmacology*; D008297:Male; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D003470:Culture Media; D005296:Ferrous Compounds; C020748:ferrous sulfate; D007501:Iron; D001205:Ascorbic Acid