Transcriptional Regulation of Jaw Osteoblasts: Development to Pathology.

Craniofacial and jaw bones have unique physiological specificities when compared to axial and appendicular bones. However, the molecular profile of the jaw osteoblast (OB) remains incomplete. The present study aimed to decipher the bone site-specific profiles of transcription factors (TFs) expressed in OBs in vivo. Using RNA sequencing analysis, we mapped the transcriptome of confirmed OBs from 2 different skeletal sites: mandible (Md) and tibia (Tb). The OB transcriptome contains 709 TF genes: 608 are similarly expressed in Md-OB and Tb-OB, referred to as "OB-core"; 54 TF genes are upregulated in Md-OB, referred to as "Md-set"; and 18 TF genes are upregulated in Tb-OB, referred to as "Tb-set." Notably, the expression of 29 additional TF genes depends on their RNA transcript variants. TF genes with no previously known role in OBs and bone were identified. Bioinformatics analysis combined with review of genetic disease databases and a comprehensive literature search showed a significant contribution of anatomical origin to the OB signatures. Md-set and Tb-set are enriched with site-specific TF genes associated with development and morphogenesis (neural crest vs. mesoderm), and this developmental imprint persists during growth and homeostasis. Jaw and tibia site-specific OB signatures are associated with craniofacial and appendicular skeletal disorders as well as neurocristopathies, dental disorders, and digit malformations. The present study demonstrates the feasibility of a new method to isolate pure OB populations and map their gene expression signature in the context of OB physiological environment, avoiding in vitro culture and its associated biases. Our results provide insights into the site-specific developmental pathways governing OBs and identify new major OB regulators of bone physiology. We also established the importance of the OB transcriptome as a prognostic tool for human rare bone diseases to explore the hidden pathophysiology of craniofacial malformations, among the most prevalent congenital defects in humans.

Cytotoxicity of glutaraldehyde and formaldehyde in relation to time of exposure and concentration.

The interacting effects of time of exposure and concentration as factors in cytotoxicity were compared for glutaraldehyde and formaldehyde. Cells from a human fibroblast cell line (WI-38) grown to confluence in 24-well trays were exposed to a range of concentrations of each agent, for periods of 4 to 24 hr. Cytotoxicity was measured by its effects on mitochondrial dehydrogenase activity, as assayed biochemically. Cytotoxic effects of formaldehyde occurred over a narrow concentration range from nontoxic to maximally toxic, and the range was little affected by time of exposure. In contrast, glutaraldehyde exerted its effect over a wider concentration range, and longer exposure times were necessary for maximal toxicity. The data suggest that long contact times of glutaraldehyde with dental pulp are necessary for maximum fixation. While 19% formaldehyde appeared to be more toxic than 2.5% glutaraldehyde in terms of serial dilution, little difference in cytotoxicity was observed when the data were calculated in terms of molar concentrations of the two agents.