Spatial distribution of mineralized bone matrix produced by marrow mesenchymal stem cells in self-assembling peptide hydrogel scaffold.
We evaluated the osteogenic differentiation of mesenchymal stem cells (MSCs) using a new class of synthetic self-assembling peptide hydrogels, RADA 16, as a scaffold for three-dimensional culture. MSCs derived from rat bone marrow were culture-expanded and seeded into the hydrogel and further cultured in osteogenic medium containing beta-glycerophosphate, ascorbic acid, and dexamethasone for 2-4 weeks. High alkaline phosphatase activity and osteocalcin (OC) contents were detected at both the protein and gene expression levels during the culture periods. Both calcium and the OC contents increased over time, indicating the growth of a mineralized extracellular matrix within the hydrogel. Moreover, the process of the growth of the mineralized matrix determined by three-dimensional microarchitecture images was obtained by confocal laser scanning microscopy. The findings show that MSCs can differentiate into mature osteoblasts to form mineralized matrices within the hydrogel scaffold. Importantly, the differentiation can occur three dimensionally within the hydrogel, indicating that RADA 16 can be considered attractive synthetic biomaterial for use in bone tissue engineering.
Hamada
Kazunori
K
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology, 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974, Japan.
Hirose
Motohiro
M
Yamashita
Toshihiko
T
Ohgushi
Hajime
H
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015519:Bone Density / Q000187:drug effects*; D001857:Bone Matrix / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005452:Fluoresceins / Q000378:metabolism; D020100:Hydrogels / Q000737:chemistry*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D015675:Osteocalcin / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011916:Rats, Inbred F344
D005452:Fluoresceins; D020100:Hydrogels; D010455:Peptides; D012333:RNA, Messenger; D015675:Osteocalcin; D000469:Alkaline Phosphatase; C007740:fluorexon