Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix

• Published in: Cell and tissue research 2015-10, Vol.362 (1), p.115-126
• Main Authors: Ro, Hyunuk, Park, Jungha, Yang, Kisuk, Kim, Jiyong, Yim, Hyun-Gu, Jung, Giyoung, Lee, Hyukjin, Cho, Seung-Woo, Hwang, Nathaniel S
• Format: Article
• Language: English
• Subjects: alizarin; alkaline phosphatase; Analysis; animal models; Animals; Biomedical and Life Sciences; Biomedicine; body fluids; bone formation; Bones; Calcification, Physiologic - physiology; cartilage; Cell Differentiation; Cellular biology; Chondrocytes - metabolism; Collagen; exposure; genes; Human Genetics; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mice; Mice, Inbred BALB C; mineralization; Molecular Medicine; osteocalcin; Osteogenesis - physiology; Proteomics; Regular Article; Rodents; Signal transduction; skull; Stem cells; Tissue Engineering; tissue repair
• ISSN: 0302-766X; 1432-0878
• DOI: 10.1007/s00441-015-2195-7

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.