Donor's age and replicative senescence favour the in-vitro mineralization potential of human fibroblasts.
Aberrant mineralization of soft connective tissues (ectopic calcification) may occur as a frequent age-related complication. Still, it remains unclear the role of mesenchymal cell donor's age and of replicative senescence on ectopic calcification. Therefore, the ability of cells to deposit in-vitro hydroxyapatite crystals and the expression of progressive ankylosis protein homolog (ANKH), ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), tissue non specific alkaline phosphatase (TNAP) and osteopontin (OPN) have been evaluated in human dermal fibroblasts derived from neonatal (nHDF) and adult (aHDF) donors (ex-vivo ageing model) or at low and high cumulative population doublings (CPD) up to replicative senescence (in-vitro ageing model). This study demonstrates that: 1) replicative senescence favours hydroxyapatite formation in cultured fibroblasts; 2) donor's age acts as a major modulator of the mineralizing potential of HDF, since nHDF are less prone than aHDF to induce calcification; 3) donor's age and replicative senescence play in concert synergistically increasing the calcification process; 4) the ANKH+ENPP1/TNAP ratio, being crucial for pyrophosphate/inorganic phosphate balance, is greatly influenced by donor's age, as well as by replicative senescence, and regulates mineral deposition; 5) OPN is only modulated by replicative senescence.
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Federica
F
Boraldi
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Angelica
A
Bartolomeo
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Caterina
C
Di Bari
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy.
Andrea
A
Cocconi
Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy. Electronic address: daniela.quaglino@unimore.it.
Daniela
D
Quaglino
D000328:Adult; D000367:Age Factors; D000469:Alkaline Phosphatase / Q000378:metabolism*; D002113:Calcification, Physiologic; D002478:Cells, Cultured; D016922:Cellular Senescence; D011756:Diphosphates / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D053495:Osteopontin / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D011755:Pyrophosphatases / Q000378:metabolism*
D011756:Diphosphates; D010710:Phosphates; C506895:SPP1 protein, human; D053495:Osteopontin; C107241:diphosphoric acid; D000469:Alkaline Phosphatase; D010727:Phosphoric Diester Hydrolases; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; D011755:Pyrophosphatases
Ageing models; Cell culture; Ectopic calcification; Mesenchymal cell