Regulation of human mesenchymal stem cells differentiation into chondrocytes in extracellular matrix-based hydrogel scaffolds.
To induce human mesenchymal stem cells (hMSCs) to differentiate into chondrocytes in three-dimensional (3D) microenvironments, we developed porous hydrogel scaffolds using the cartilage extracellular matrix (ECM) components of chondroitin sulfate (CS) and collagen (COL). The turbidity and viscosity experiments indicated hydrogel could form through pH-triggered co-precipitation when pH=2-3. Enzyme-linked immunosorbent assay (ELISA) confirmed the hydrogel scaffolds could controllably release growth factors as envisaged. Transforming growth factor-β (TGF-β) was released to stimulate hMSCs differentiation into chondrocytes; and then collagen binding domain-basic fibroblast growth factor (CBD-bFGF) was released to improve the differentiation and preserve the chondrocyte phenotype. In in vitro cell culture experiments, the differentiation processes were compared in different microenvironments: 2D culture in culture plate as control, 3D culture in the fabricated scaffolds without growth factors (CC), the samples with CBD-bFGF (CC-C), the samples with TGF-β (CC-T), the samples with CBD-bFGF/TGF-β (CC-CT). Real-time polymerase chain reaction (RT-PCR) revealed the hMSC marker genes of CD44 and CD105 decreased; at the same time the chondrocyte marker genes of collagen type II and aggrecan increased, especially in the CC-CT sample. Immunostaining results further confirmed the hMSC marker protein of CD 44 disappeared and the chondrocyte marker protein of collagen type II emerged over time in the CC-CT sample. These results imply the ECM-based hydrogel scaffolds with growth factors can supply suitable 3D cell niches for hMSCs differentiation into chondrocytes and the differentiation process can be regulated by the controllably released growth factors.
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Mingchun
M
Du
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Hui
H
Liang
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Chenchen
C
Mou
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Xiaoran
X
Li
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Jie
J
Sun
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China.
Yan
Y
Zhuang
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), Beijing 100190, PR China.
Zhifeng
Z
Xiao
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), Beijing 100190, PR China.
Bing
B
Chen
Division of Nanobiomedicine Research and Suzhou Key Laboratory of Nanobiomedical Characterization, Suzhou Institute of Nano-Tech and Nano-Bionics (SINANO), Chinese Academy of Sciences (CAS), 398 Ruoshui Road, Suzhou Dushu Lake Science and Education Innovation District (SEID), Suzhou Industrial Park (SIP), Suzhou 215123, PR China; State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences (CAS), Beijing 100190, PR China. Electronic address: jwdai@genetics.ac.cn.
Jianwu
J
Dai
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D013194:Staining and Labeling; D054457:Tissue Scaffolds / Q000737:chemistry*; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D015415:Biomarkers; D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
Chondrocyte; Controlled release; Differentiation; Extracellular matrix; Growth factor; Human mesenchymal stem cell