Bone differentiation of marrow-derived mesenchymal stem cells using beta-tricalcium phosphate-alginate-gelatin hybrid scaffolds.
The aim of the present study was to establish a 3D culture system for bone differentiation of mesenchymal stem cells (MSCs), using a new hybrid sponge. To manufacture the scaffold, a composite of beta-tricalcium phosphate-alginate-gelatin was prepared and cast as pellets of 1 cm diameter. The sponge was then fabricated by drying in freeze-dryer for 12 h. The porosity, mean pore size, compressive modulus and strength of the composite sponge fabricated in this study were 89.7%, 325.3 microm, 1.82 and 0.196 MPa, respectively. To establish a 3D culture system, the rat bone marrow-derived MSCs were suspended in 500 microl diluted collagen gel, loaded into the porous sponge and provided with medium with or without osteogenic supplements for 3 weeks. The day after loading, the cells appeared in the scaffold's internal spaces, where later some of them from either culture survived by anchoring on the surfaces. At the end of cultivation period, individually adhered cells from both cultures were observed to be replaced by cell aggregates, in which mineralized matrix was detected by alizarin red staining. Furthermore, RT-PCR analysis indicated that the bone-specific gene osteocalcin was expressed in cultures in both the presence and absence of the osteogenic supplements. Taken together, it seems that the studied scaffolds are cell-compatible and, more importantly, possess some osteo-inductive properties.
Eslaminejad
Mohamadreza Baghaban
MB
Stem Cell Department, Royan Institute, PO Box 19395-4644, Tehran, Iran. bagesla@yahoo.com
Mirzadeh
Hamid
H
Mohamadi
Yossef
Y
Nickmahzar
Aghbibi
A
D000464:Alginates / Q000378:metabolism*; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D018786:Bone Substitutes; D001842:Bone and Bones / Q000166:cytology*; D002130:Calcium Phosphates / Q000378:metabolism*; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D005780:Gelatin / Q000378:metabolism*; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D016062:Porosity; D051381:Rats; D017208:Rats, Wistar; D054457:Tissue Scaffolds / Q000737:chemistry*
D000464:Alginates; D018786:Bone Substitutes; D002130:Calcium Phosphates; D006603:Hexuronic Acids; C485817:beta-tricalcium phosphate; D020723:Glucuronic Acid; D005780:Gelatin