Development of thick and highly cell-incorporated engineered tissues by hydrogel template approach with basic fibroblast growth factor or ascorbic acid.
We have previously reported a hydrogel template approach for the construction of centimeter-sized three-dimensional (3D) engineered tissues composed of cultured cells and extracellular matrices (ECM) produced by the cells. However, the interior of the engineered tissues was low in cell density; thus, it was pouched and non-dense morphologies. In this study, we developed thick and highly cell-incorporated engineered tissues by using basic fibroblast growth factor (bFGF) and ascorbic acid 2-phosphate (Asc 2-P). bFGF was loaded into freeze-dried poly(gamma-glutamic acid) hydrogels with disulfide cross-links (gamma-PGA-SS gels) as a template to enhance cell growth. After prescribed times of cell culture, the scaffolds were decomposed by adding a biocompatible reductant, cysteine, and cell proliferation and collagen production were investigated. The loading of bFGF into the scaffolds enhanced cellular invasion and proliferation. Meanwhile, the addition of Asc 2-P to the culture medium induced collagen production from the adhered fibroblasts. However, Asc 2-P did not affect cellular invasion into the template gamma-PGA-SS gels. The volume and weight of the obtained tissues using bFGF after 28 days of culture were 1.3- and 1.4-fold higher than those of control tissues. The thick and highly cell-incorporated 3D-engineered tissues can be useful as a novel cell implantation material for tissue engineering.
Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita 565-0871, Japan.
Hiroaki
H
Yoshida
Michiya
M
Matsusaki
Mitsuru
M
Akashi
D001205:Ascorbic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism / Q000648:ultrastructure; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000378:metabolism*; D011099:Polyglutamic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
C511775:poly(gamma-glutamic acid); D016222:Fibroblast Growth Factor 2; C011669:ascorbate-2-phosphate; D011099:Polyglutamic Acid; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D003094:Collagen; D001205:Ascorbic Acid