Bone tissue engineering using human adipose‐derived stem cells and honeycomb collagen scaffold

Abstract

Human adipose-derived stem cells (ASCs) have the capacity to regenerate and the potential to differentiate into multiple lineages of mesenchymal cells. The aim of this study was to investigate the possibility of using honeycomb collagen scaffold to culture ASCs in bone tissue engineering. The osteogenic capacity of ASCs in vitro, was confirmed by histology and measuring the expression of cbfa-1. After that, ASCs were cultured for up to 14 days in the honeycomb scaffold to allow a high density, three-dimensional culture. Scanning electron microscopy data showed that the scaffold was filled with the grown ASCs, and calcification, stained black with von Kossa, was confirmed. Furthermore, The ASC-loaded honeycomb collagen scaffolds cultured for 14 days were subcutaneously transplanted into nude mice, and excised after 8 weeks. Bone formation in vivo was examined using HE stain, von Kossa stain, and osteocalcin immunostain. Those histological views showed significant positive stains in the samples of osteogenic medium in the three types of stain. These results suggest that this carrier is a suitable scaffold for ASCs and will be useful as a three-dimensional bone tissue engineering scaffold in vitro and in vivo.

Review of biophysical factors affecting osteogenic differentiation of human adult adipose-derived stem cells

“…Hattori and colleagues describe better osteoconductivity on a β-TCP scaffold for ASCs, as compared with scaffolds of hydroxyapatite (HA) and atelocollagen honeycomb scaffolds (Hattori et al 2006). Kakudo and colleagues showed better osteogenesis in cells cultured with osteogenic chemical supplements as compared to control growth medium (Kakudo et al 2008). Taken together, the results of these studies suggest that β-TCP is a better osteogenic scaffold material for ASCs than HA and that atelocollagen scaffolds hold promise for ASC bone engineering applications where their low mechanical strength is acceptable.…”

“…For evaluation of in vivo osteogenesis, an immunohistochemistry assay was performed to identify osteocalcin in histological sections after 2 weeks culture in osteogenic medium followed by 8 weeks subcutaneous implantation in mice. To gauge osteogenic capacity in vitro, Kakudo and colleagues assessed mineralization and alkaline phosphatase activity at day 14 after induction, also measuring calcification with scanning electron microscopy (Kakudo et al 2008). This group used qRT-PCR to verify increase in expression of osteogenic marker transcription factor Runx2 with respect to control at days 7 and 14 after induction.…”

Mesenchymal Stem Cells and Tooth Engineering

“…It showed that the scaffold was filled with the grown ADSCs and calcification materials. When the ADSCs-loaded honeycomb collagen scaffolds were subcutaneously transplanted into nude mice, bone formation in vivo was identified after 8 weeks (Kakudo et al, 2008).…”

The Role of the Wnt Signaling Pathway in the Osteogenic Differentiation of Human Adipose-derived Stem Cells under Mechanical Stimulation

“…Recently, many studies have demonstrated that hASCs represent an important new cell source for bone tissue engineering [11][12][13] .…”