In vivo osteogenic differentiation of human turbinate mesenchymal stem cells in an injectable in situ-forming hydrogel.
Human turbinate mesenchymal stromal cells (hTMSCs) are an alternate source of adult stem cells for regenerative medicine. In this work, we demonstrated that hTMSCs are easily harvested from turbinate tissue using a minimal surgical procedure. hTMSCs showed positive expression of mesenchymal stem cell markers and proliferated at a high rate. The specific surface proteins of harvested hTMSCs were relatively tolerant of ex vivo manipulation in culture. hTMSCs exhibited osteogenic differentiation in vitro in the presence of osteogenic factors. To examine osteogenic differentiation of hTMSCs in vivo in an injectable hydrogel, cells were incorporated into a methoxy polyethylene glycol-polycaprolactone block copolymer (MPEG-PCL (MP)) solution simply by mixing. hTMSC-loaded MP solutions exhibited a temperature-dependent solution-to-gel phase transition. The hTMSC attached and grew well on in vitro- and in vivo-formed MP hydrogels. hTMSC-loaded MP solutions formed a hydrogel almost immediately upon injection into animals and the cells remained viable, even after 12 weeks. Injected hTMSCs in in situ-formed MP hydrogels differentiated into osteogenic cells, mainly in the presence of osteogenic factors. Differentiated osteoblasts were identified by Alizarin Red S, von Kossa, and alkaline phosphatase (ALP) staining, and osteonectin, osteopontin, and osteocalcin mRNA expression. To the best of our knowledge, this is the first study to show hTMSCs undergoing osteogenic differentiation in in vivo-formed MP hydrogels. In conclusion, hTMSCs could serve as adult stem cell sources and, when embedded in an in situ-formed hydrogel, may provide numerous benefits as a noninvasive alternative for bone tissue engineering applications.
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
Jin Seon
JS
Kwon
Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul 137-701, Republic of Korea.
Sung Won
SW
Kim
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
Doo Yeon
DY
Kwon
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
Seung Hun
SH
Park
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
A Reum
AR
Son
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
Jae Ho
JH
Kim
Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea. Electronic address: moonskim@ajou.ac.kr.
Moon Suk
MS
Kim
D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D015870:Gene Expression; D006801:Humans; D020100:Hydrogels / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D008855:Microscopy, Electron, Scanning; D010012:Osteogenesis; D044968:Regenerative Medicine; D023822:Tissue Engineering; D014420:Turbinates / Q000166:cytology*
D020100:Hydrogels
Injectable hydrogel; Osteogenic differentiation; Tissue engineering; Turbinate mesenchymal stem cells