Age-related decline in the matrix contents and functional properties of human periodontal ligament stem cell sheets.
In this study, periodontal ligament (PDL) stem cells (PDLSCs) derived from different-aged donors were used to evaluate the effect of aging on cell sheet formation. The activity of PDLSCs was first determined based on their colony-forming ability, surface markers, proliferative/differentiative potentials, senescence-associated β-galactosidase (SA-βG) staining, and expression of pluripotency-associated transcription factors. The ability of these cells to form sheets, based on their extracellular matrix (ECM) contents and their functional properties necessary for osteogenic differentiation, was evaluated to predict the age-related changes in the regenerative capacity of the cell sheets in their further application. It was found that human PDLSCs could be isolated from the PDL tissue of different-aged subjects. However, the ability of the PDLSCs to proliferate and to undergo osteogenic differentiation and their expression of pluripotency-associated transcription factors displayed age-related decreases. In addition, these cells exhibited an age-related increase in SA-βG expression. Aged cells showed an impaired ability to form functional cell sheets, as determined by morphological observations and Ki-67 immunohistochemistry staining. Based on the production of ECM proteins, such as fibronectin, integrin β1, and collagen type I; alkaline phosphatase (ALP) activity; and the expression of osteogenic genes, such as ALP, Runt-related transcription factor 2, and osteocalcin, cell sheets formed by PDLSCs derived from older donors demonstrated a less potent osteogenic capacity compared to those formed by PDLSCs from younger donors. Our data suggest that the age-associated decline in the matrix contents and osteogenic properties of PDLSC sheets should be taken into account in cell sheet engineering research and clinical periodontal regenerative therapy.
State Key Laboratory of Military Stomatology Biomaterials Unit, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, PR China.
Rui-Xin
RX
Wu
State Key Laboratory of Military Stomatology Biomaterials Unit, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, PR China.
Chun-Sheng
CS
Bi
State Key Laboratory of Military Stomatology Biomaterials Unit, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, PR China.
Yang
Y
Yu
State Key Laboratory of Military Stomatology Biomaterials Unit, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, PR China.
Lin-Lin
LL
Zhang
State Key Laboratory of Military Stomatology Biomaterials Unit, Department of Periodontology, School of Stomatology, Fourth Military Medical University, Xi'an, PR China. Electronic address: cfmsunhh@fmmu.edu.cn.
Fa-Ming
FM
Chen
D050156:Adipogenesis; D000293:Adolescent; D000328:Adult; D000375:Aging / Q000502:physiology*; D015415:Biomarkers / Q000378:metabolism; D002113:Calcification, Physiologic / Q000235:genetics; D049109:Cell Proliferation; D048430:Cell Shape; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D016130:Immunophenotyping; D008297:Male; D008875:Middle Aged; D010012:Osteogenesis; D010513:Periodontal Ligament / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D055815:Young Adult; D001616:beta-Galactosidase / Q000378:metabolism
D015415:Biomarkers; D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D001616:beta-Galactosidase
Cell sheet engineering; Matrix production; Osteogenic differentiation; Periodontal regeneration; Stem cell aging