Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Background: The developing periodontium is sensitive to local levels of inorganic phosphate (P(i)) and inorganic pyrophosphate (PP(i)) as demonstrated by cementum phenotypes resulting from the loss of function of protein regulators of P(i)/PP(i) homeostasis. The progressive ankylosis protein (ANK) regulates the transport of PP(i), and progressive ankylosis gene (Ank) and knock-out (KO) mice feature a rapidly forming and thick cementum. We hypothesized that, besides affecting cementum formation, decreased extracellular PP(i) levels in Ank KO mice would also impact cementum regeneration. Methods: Periodontal fenestration defects (approximate to 2 mm in length, 1 mm in width, and 0.5 mm in depth) were created on buccal aspects of mandibular molars in Ank KO and wild-type (WT) mice. Mandibles were harvested at 15 and 30 days post-surgery for histology, histomorphometry, evaluation of in vivo fluorochrome labeling, and immunohistochemistry (IHC) for proteins including bone sialoprotein (BSP), osteopontin (OPN), dentin matrix protein 1 (DMP1), and ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1). Results: A greater amount of new cementum was observed in Ank KO mice at 15 and 30 days post-surgery (P < 0.05), which was confirmed by fluorochrome labeling showing a higher new cementum appositional activity in defect areas in Ank KO mice versus controls. At days 15 and 30 during healing, regenerating cementum and associated cells in Ank KO samples recapitulated expression patterns mapped during development, including limited BSP and positive OPN and DMP1 in the cementum matrix as well as elevated NPP1 in cemento-blasts. Conclusions: Within the limits of the study, these findings suggest that reduced local levels of PP, could promote increased cementum regeneration. Therefore, the local modulation of P(i)/PP(i) may be a potential therapeutic approach for achieving improved cementum regeneration. J Periodontol 2011;82: 1757-1766.821217571766Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)National Institutes of Health, Bethesda, Maryland [5R03TW007590-03]National Institute of Dental and Craniofacial Research (NIDCR) [DE15109]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [08/00534-7]National Institutes of Health, Bethesda, Maryland [5R03TW007590-03]National Institute of Dental and Craniofacial Research (NIDCR) [DE15109]