Abstract
Transient cartilage and a mineralizing microenvironment play pivotal roles in mesenchymal cell ossification during bone formation. In order to recreate these microenvironmental cues, C3H10T1/2 murine mesenchymal stem cells (MSCs) were exposed to chondrocyte-conditioned medium (CM) and seeded onto three-dimensional mineralized scaffolds for bone regeneration. Expansion of C3H10T1/2 cells with CM resulted in enhanced expression levels of chondrogenic markers such as aggrecan, type II collagen, type X collagen, and Sox9, rather than of osteogenic genes. Interestingly, CM expansion led to reduced expression levels of osteogenic genes such as alkaline phosphatase (ALP), type I collagen, osteocalcin, and Runx2. However, CM-expanded C3H10T1/2 cells showed enhanced osteogenic differentiation as indicated by increased ALP and Alizarin Red S staining upon osteogenic factor exposure. In vivo, CM-expanded C3H10T1/2 mesenchymal cells were seeded onto mineralized scaffolds (fabricated with polydopamine and coated with simulated body fluids) and implanted into critical-sized calvarial-defect mouse models. After 8 weeks of implantation, mouse skulls were collected, and bone tissue regeneration was evaluated by micro-computed tumography and Masson’s trichrome staining. In accordance with the in vitro analysis, CM-expanded C3H10T1/2 cells gave enhanced bone mineral deposition. Thus, chondrocyte-conditioned factors and a mineralized microenvironment stimulate the bone formation of MSCs.
| Original language | English |
|---|---|
| Pages (from-to) | 115-126 |
| Number of pages | 12 |
| Journal | Cell and Tissue Research |
| Volume | 362 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2015 Oct 22 |
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All Science Journal Classification (ASJC) codes
- Pathology and Forensic Medicine
- Histology
- Cell Biology
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Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix. / Ro, Hyunuk; Park, Jungha; Yang, Kisuk; Kim, Jiyong; Yim, Hyun Gu; Jung, Giyoung; Lee, Hyukjin; Cho, Seung Woo; Hwang, Nathaniel S.
In: Cell and Tissue Research, Vol. 362, No. 1, 22.10.2015, p. 115-126.Research output: Contribution to journal › Article
TY - JOUR
T1 - Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix
AU - Ro, Hyunuk
AU - Park, Jungha
AU - Yang, Kisuk
AU - Kim, Jiyong
AU - Yim, Hyun Gu
AU - Jung, Giyoung
AU - Lee, Hyukjin
AU - Cho, Seung Woo
AU - Hwang, Nathaniel S.
PY - 2015/10/22
Y1 - 2015/10/22
N2 - Transient cartilage and a mineralizing microenvironment play pivotal roles in mesenchymal cell ossification during bone formation. In order to recreate these microenvironmental cues, C3H10T1/2 murine mesenchymal stem cells (MSCs) were exposed to chondrocyte-conditioned medium (CM) and seeded onto three-dimensional mineralized scaffolds for bone regeneration. Expansion of C3H10T1/2 cells with CM resulted in enhanced expression levels of chondrogenic markers such as aggrecan, type II collagen, type X collagen, and Sox9, rather than of osteogenic genes. Interestingly, CM expansion led to reduced expression levels of osteogenic genes such as alkaline phosphatase (ALP), type I collagen, osteocalcin, and Runx2. However, CM-expanded C3H10T1/2 cells showed enhanced osteogenic differentiation as indicated by increased ALP and Alizarin Red S staining upon osteogenic factor exposure. In vivo, CM-expanded C3H10T1/2 mesenchymal cells were seeded onto mineralized scaffolds (fabricated with polydopamine and coated with simulated body fluids) and implanted into critical-sized calvarial-defect mouse models. After 8 weeks of implantation, mouse skulls were collected, and bone tissue regeneration was evaluated by micro-computed tumography and Masson’s trichrome staining. In accordance with the in vitro analysis, CM-expanded C3H10T1/2 cells gave enhanced bone mineral deposition. Thus, chondrocyte-conditioned factors and a mineralized microenvironment stimulate the bone formation of MSCs.
AB - Transient cartilage and a mineralizing microenvironment play pivotal roles in mesenchymal cell ossification during bone formation. In order to recreate these microenvironmental cues, C3H10T1/2 murine mesenchymal stem cells (MSCs) were exposed to chondrocyte-conditioned medium (CM) and seeded onto three-dimensional mineralized scaffolds for bone regeneration. Expansion of C3H10T1/2 cells with CM resulted in enhanced expression levels of chondrogenic markers such as aggrecan, type II collagen, type X collagen, and Sox9, rather than of osteogenic genes. Interestingly, CM expansion led to reduced expression levels of osteogenic genes such as alkaline phosphatase (ALP), type I collagen, osteocalcin, and Runx2. However, CM-expanded C3H10T1/2 cells showed enhanced osteogenic differentiation as indicated by increased ALP and Alizarin Red S staining upon osteogenic factor exposure. In vivo, CM-expanded C3H10T1/2 mesenchymal cells were seeded onto mineralized scaffolds (fabricated with polydopamine and coated with simulated body fluids) and implanted into critical-sized calvarial-defect mouse models. After 8 weeks of implantation, mouse skulls were collected, and bone tissue regeneration was evaluated by micro-computed tumography and Masson’s trichrome staining. In accordance with the in vitro analysis, CM-expanded C3H10T1/2 cells gave enhanced bone mineral deposition. Thus, chondrocyte-conditioned factors and a mineralized microenvironment stimulate the bone formation of MSCs.
UR - http://www.scopus.com/inward/record.url?scp=84941877298&partnerID=8YFLogxK
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U2 - 10.1007/s00441-015-2195-7
DO - 10.1007/s00441-015-2195-7
M3 - Article
C2 - 25956591
AN - SCOPUS:84941877298
VL - 362
SP - 115
EP - 126
JO - Cell and Tissue Research
JF - Cell and Tissue Research
SN - 0302-766X
IS - 1
ER -