Direkt zum Inhalt
Merck

Periodontal Defects in the A116T Knock-in Murine Model of Odontohypophosphatasia.

Journal of dental research (2015-02-27)
B L Foster, C R Sheen, N E Hatch, J Liu, E Cory, S Narisawa, T Kiffer-Moreira, R L Sah, M P Whyte, M J Somerman, J L Millán
ZUSAMMENFASSUNG

Mutations in ALPL result in hypophosphatasia (HPP), a disease causing defective skeletal mineralization. ALPL encodes tissue nonspecific alkaline phosphatase (ALP), an enzyme that promotes mineralization by reducing inorganic pyrophosphate, a mineralization inhibitor. In addition to skeletal defects, HPP causes dental defects, and a mild clinical form of HPP, odontohypophosphatasia, features only a dental phenotype. The Alpl knockout (Alpl (-/-)) mouse phenocopies severe infantile HPP, including profound skeletal and dental defects. However, the severity of disease in Alpl (-/-) mice prevents analysis at advanced ages, including studies to target rescue of dental tissues. We aimed to generate a knock-in mouse model of odontohypophosphatasia with a primarily dental phenotype, based on a mutation (c.346G>A) identified in a human kindred with autosomal dominant odontohypophosphatasia. Biochemical, skeletal, and dental analyses were performed on the resulting Alpl(+/A116T) mice to validate this model. Alpl(+/A116T) mice featured 50% reduction in plasma ALP activity compared with wild-type controls. No differences in litter size, survival, or body weight were observed in Alpl(+/A116T) versus wild-type mice. The postcranial skeleton of Alpl(+/A116T) mice was normal by radiography, with no differences in femur length, cortical/trabecular structure or mineral density, or mechanical properties. Parietal bone trabecular compartment was mildly altered. Alpl(+/A116T) mice featured alterations in the alveolar bone, including radiolucencies and resorptive lesions, osteoid accumulation on the alveolar bone crest, and significant differences in several bone properties measured by micro-computed tomography. Nonsignificant changes in acellular cementum did not appear to affect periodontal attachment or function, although circulating ALP activity was correlated significantly with incisor cementum thickness. The Alpl(+/A116T) mouse is the first model of odontohypophosphatasia, providing insights on dentoalveolar development and function under reduced ALP, bringing attention to direct effects of HPP on alveolar bone, and offering a new model for testing potential dental-targeted therapies in future studies.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Essigsäure, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Essigsäure, glacial, ReagentPlus®, ≥99%
Sigma-Aldrich
Formaldehyd -Lösung, Molecular Biology, 36.5-38% in H2O
Sigma-Aldrich
Molybdatophosphorsäure Hydrat, ACS reagent
Sigma-Aldrich
Natriumchlorid, Molecular Biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Formaldehyd -Lösung, ACS reagent, 37 wt. % in H2O, contains 10-15% Methanol as stabilizer (to prevent polymerization)
SAFC
Formaldehyd -Lösung, contains 10-15% methanol as stabilizer, 37 wt. % in H2O
Sigma-Aldrich
Natriumchlorid -Lösung, 5 M in H2O, BioReagent, Molecular Biology
Sigma-Aldrich
Essigsäure, glacial, ≥99.99% trace metals basis
Sigma-Aldrich
Natriumchlorid, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Natriumchlorid, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Natriumchlorid -Lösung, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Essigsäure -Lösung, suitable for HPLC
Sigma-Aldrich
Pikrinsäure, moistened with water, ≥98%
Sigma-Aldrich
Natriumchlorid -Lösung, 5 M
SAFC
Natriumchlorid -Lösung, 5 M
Sigma-Aldrich
Formaldehyd -Lösung, Molecular Biology, BioReagent, ≥36.0% in H2O (T)
Sigma-Aldrich
Pikrinsäure -Lösung, 1.3% in H2O (saturated)
Sigma-Aldrich
Natriumchlorid, BioUltra, Molecular Biology, ≥99.5% (AT)
Sigma-Aldrich
Natriumchlorid, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%
Sigma-Aldrich
Natriumchlorid -Lösung, BioUltra, Molecular Biology, ~5 M in H2O
Sigma-Aldrich
Natriumchlorid, 99.999% trace metals basis
Sigma-Aldrich
Essigsäure, suitable for luminescence, BioUltra, ≥99.5% (GC)
Supelco
Natriumchlorid, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Eisessig, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Formaldehyd -Lösung, meets analytical specification of USP, ≥34.5 wt. %
Sigma-Aldrich
5α-Androstan-17β-ol-3-on, ≥97.5%
Supelco
Natriumchlorid, reference material for titrimetry, certified by BAM, >99.5%
Supelco
Formaldehyd -Lösung, stabilized with methanol, ~37 wt. % in H2O, certified reference material
Sigma-Aldrich
3-Amino-9-ethylcarbazol, tablet