title: Osteogenic priming of mesenchymal stem cells by chondrocyte-conditioned factors and mineralized matrix.
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.
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Hyunuk
authors.initials: H
authors.lastname: Ro
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Jungha
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul, 120-749, Republic of Korea.
authors.forename: Kisuk
authors.initials: K
authors.lastname: Yang
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Kim
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea.
authors.forename: Hyun-Gu
authors.initials: HG
authors.lastname: Yim
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea.|College of Pharmacy, Ewha Womans University, Seoul, 120-750, Republic of Korea.
authors.forename: Giyoung
authors.initials: G
authors.lastname: Jung
authors.affiliation: College of Pharmacy, Ewha Womans University, Seoul, 120-750, Republic of Korea.
authors.forename: Hyukjin
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul, 120-749, Republic of Korea.
authors.forename: Seung-Woo
authors.initials: SW
authors.lastname: Cho
authors.affiliation: School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, Seoul, 151-742, Republic of Korea. nshwang@snu.ac.kr.|The NBio Institute of Seoul National University, Seoul, 151-742, Republic of Korea. nshwang@snu.ac.kr.
authors.forename: Nathaniel S
authors.initials: NS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D002113:Calcification, Physiologic / Q000502:physiology*; D002454:Cell Differentiation; D019902:Chondrocytes / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010012:Osteogenesis / Q000502:physiology*; D023822:Tissue Engineering
keywords: Conditioned medium; In vivo implantation; Mineralized scaffolds; Mouse (BALB/c); Osteogenic differentiation; Simulated body fluids; Stem cells; Tissue engineering