Human recombinant cementum attachment protein (hrPTPLa/CAP) promotes hydroxyapatite crystal formation in vitro and bone healing in vivo
Por:
Montoya G., Arenas J., Romo E., Zeichner-David M., Alvarez M., Narayanan A.S., Velázquez U., Mercado G., Arzate H.
Publicada:
1 dic 2014
Resumen:
Cementum extracellular matrix is similar to other mineralized tissues;
however, this unique tissue contains molecules only present in cementum.
A cDNA of these molecules, cementum attachment protein (hrPTPLa/CAP) was
cloned and expressed in a prokaryotic system. This molecule is an
alternative splicing of protein tyrosine phosphatase-like A (PTPLa). In
this study, we wanted to determine the structural and functional
characteristics of this protein. Our results indicate that hrPTPLa/CAP
contains a 43.2% alpha-helix, 8,9%beta-sheet, 2%beta-turn and 45.9%
random coil secondary structure. Dynamic light scattering shows that
this molecule has a size distribution of 4.8 nm and aggregates as an
estimated mass of 137 kDa species. AFM characterization and FE-SEM
studies indicate that this protein self-assembles into nanospheres with
sizes ranging from 7.0 to 27 nm in diameter. Functional studies
demonstrate that hrPTPLa/CAP promotes hydroxyapatite crystal nucleation:
EDS analysis revealed that hrPTPLa/CAP-induced crystals had a 1.59 +/-
0.06 Ca/P ratio. Further confirmation with MicroRaman spectrometry and
TEM confirm the presence of hydroxyapatite. In vivo studies using
critical-size defects in rat cranium showed that hrPTPLa/CAP promoted
73% +/- 2.19% and 87% +/- 1.97% new bone formation at 4 and 8 weeks
respectively. Although originally identified in cementum, PTPLa/CAP is
very effective at inducing bone repair and healing and therefore this
novel molecule has a great potential to be used for mineralized tissue
bioengineering and tissue regeneration. (C) 2014 Elsevier Inc. All
rights reserved.
Filiaciones:
Montoya G.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Biol Periodontal, Mexico City 04510, DF, Mexico
Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
Arenas J.:
Univ Nacl Autonoma Mexico, Inst Fis, Mexico City 04510, DF, Mexico
Instituto de Física, Universidad Nacional Autónoma de México, Mexico
Romo E.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Biol Periodontal, Mexico City 04510, DF, Mexico
Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
Zeichner-David M.:
Ostrow School of Dentistry, University of Southern California, United States
Alvarez M.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Bioingn Tejidos, Mexico City 04510, DF, Mexico
Laboratorio de Bioingeniería de Tejidos, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
Narayanan A.S.:
School of Medicine, Department of Pathology, University of Washington, Seattle, United States
Velázquez U.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Biol Periodontal, Mexico City 04510, DF, Mexico
Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
Mercado G.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Biol Periodontal, Mexico City 04510, DF, Mexico
Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
Arzate H.:
Univ Nacl Autonoma Mexico, Fac Odontol, Lab Biol Periodontal, Mexico City 04510, DF, Mexico
Laboratorio de Biología Periodontal, Facultad de Odontología, Universidad Nacional Autónoma de México, Mexico
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