title: Perfusion culture enhances osteogenic differentiation of rat mesenchymal stem cells in collagen sponge reinforced with poly(glycolic Acid) fiber.
abstract: The objective of this study was to obtain fundamental knowledge about in vitro culture systems to enhance the proliferation and differentiation of mesenchymal stem cells (MSCs) in collagen sponge reinforced by the incorporation of poly(glycolic acid) (PGA) fiber. A collagen solution with PGA fiber homogeneously localized at PGA:collagen weight ratios of 0.67, 1.25, 2.5, and 5 was freezedried, followed by cross-linking of combined dehydrothermal, glutaraldehyde, and ultraviolet treatment. Scanning electron microscopy revealed that collagen sponges exhibited homogeneous and interconnected pore structures with an average size of 180 microm, irrespective of PGA fiber incorporation. When rat MSCs were seeded into collagen sponge with or without PGA fiber incorporation, more attached cells were observed in collagen sponge incorporating PGA fiber than in collagen sponge without PGA fiber incorporation, irrespective of the PGA:collagen ratio. The proliferation and osteogenic differentiation of MSCs in PGA-reinforced sponge at a weight ratio of 5 were greatly influenced by the culture method and growth conditions. Alkaline phosphatase (ALP) activity and osteocalcin content of MSCs cultured in PGA-reinforced sponge by the perfusion method became maximum at a flow rate of 0.2 mL/min, although they increased with culture time period. It may be concluded that appropriate perfusion conditions enable MSCs to positively improve the extent of proliferation and differentiation.
authors.affiliation: Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
authors.forename: Hossein
authors.initials: H
authors.lastname: Hosseinkhani
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Inatsugu
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Hiraoka
authors.forename: Sachiko
authors.initials: S
authors.lastname: Inoue
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tabata
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000528:radiation effects; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000737:chemistry* / Q000528:radiation effects; D003890:Desiccation; D005269:Femur / Q000166:cytology; D005976:Glutaral / Q000737:chemistry; D006358:Hot Temperature; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis; D010477:Perfusion / Q000379:methods*; D011100:Polyglycolic Acid / Q000737:chemistry* / Q000528:radiation effects; D016062:Porosity; D051381:Rats; D011916:Rats, Inbred F344; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods; D014466:Ultraviolet Rays
chemical_list: D001672:Biocompatible Materials; D024042:Collagen Type I; D015675:Osteocalcin; D011100:Polyglycolic Acid; D000469:Alkaline Phosphatase; D005976:Glutaral