Engineering of artificial bone is a recent approach to treat patients with bone defects. Production of a bone-like tissue in vitro is based on the ability of cells to migrate, proliferate, and differentiate in culture. The cell population called human Adipose-Derived Stem Cells (hASCs) share many of the characteristic of its counterpart of marrow including extensive proliferative potential and the ability to undergo multilineage differentiation along classical mesenchimal lineages. Under osteogenic conditions, hASCs are observed to express genes and proteins associated with the osteoblast phenotype, including alkaline phosphatase, type I collagen, osteopontin, osteocalcin, bone sialoproteins, RunX-1, BMP-2, BMP-4, BMP receptors I and II, PTH receptor. Trabecular titanium scaffold (Ti6Al4V) is an inert biomaterial with an excellent biocompatibility and a great mechanical strength. The aim of this study was to evaluate with biochemical and morphological methods the adhesion and differentiation of hASCs grown in osteogenic medium on trabecular titanium scaffolds. hASCs were positive for CD90, CD73 and CD105 surface antigens and negative for CD34 and CD45. Alkaline phosphatase activity was significantly higher (p<0.05) than in controls. RT-PCR expression of type I collagen in hASCs was higher at day 21 and decreased at day 31 whereas expression of osteopontin and osteocalcin mRNA of hASCs was observed to increase from 21 to 31 days of culture. SEM observations showed cells attached onto the surface of porous scaffold and embedded in a plentiful extracellular matrix (ECM). TEM of cells showed the presence of RER cysternae and an ECM with densely packed collagen fibres exhibiting a regular and parallel arrangement. In the differentiated constructs,the presence of calcium and phosphorus was detected by x-rays to infer that calcium phosphate had formed. In conclusion, hASCs showed to be an ideal source of cells for tissue engineering and are able to acquire an osteoblastic phenotype.
Osteogenic differentiation of human Adipose-Derived Stem Cells hASCs in trabecular titanium scaffold
GASTALDI, GIULIA;SCAFFINO, MANUELA FEDERICA;GIOGLIO, LUCIANA;BENAZZO, FRANCESCO
2009-01-01
Abstract
Engineering of artificial bone is a recent approach to treat patients with bone defects. Production of a bone-like tissue in vitro is based on the ability of cells to migrate, proliferate, and differentiate in culture. The cell population called human Adipose-Derived Stem Cells (hASCs) share many of the characteristic of its counterpart of marrow including extensive proliferative potential and the ability to undergo multilineage differentiation along classical mesenchimal lineages. Under osteogenic conditions, hASCs are observed to express genes and proteins associated with the osteoblast phenotype, including alkaline phosphatase, type I collagen, osteopontin, osteocalcin, bone sialoproteins, RunX-1, BMP-2, BMP-4, BMP receptors I and II, PTH receptor. Trabecular titanium scaffold (Ti6Al4V) is an inert biomaterial with an excellent biocompatibility and a great mechanical strength. The aim of this study was to evaluate with biochemical and morphological methods the adhesion and differentiation of hASCs grown in osteogenic medium on trabecular titanium scaffolds. hASCs were positive for CD90, CD73 and CD105 surface antigens and negative for CD34 and CD45. Alkaline phosphatase activity was significantly higher (p<0.05) than in controls. RT-PCR expression of type I collagen in hASCs was higher at day 21 and decreased at day 31 whereas expression of osteopontin and osteocalcin mRNA of hASCs was observed to increase from 21 to 31 days of culture. SEM observations showed cells attached onto the surface of porous scaffold and embedded in a plentiful extracellular matrix (ECM). TEM of cells showed the presence of RER cysternae and an ECM with densely packed collagen fibres exhibiting a regular and parallel arrangement. In the differentiated constructs,the presence of calcium and phosphorus was detected by x-rays to infer that calcium phosphate had formed. In conclusion, hASCs showed to be an ideal source of cells for tissue engineering and are able to acquire an osteoblastic phenotype.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
