Tissue engineering of cementum/periodontal-ligament complex using a novel three-dimensional pellet cultivation system for human periodontal ligament stem cells.
Limitations of conventional regeneration modalities underscore the necessity of recapitulating development for periodontal tissue engineering. In this study, we proposed a novel three-dimensional pellet cultivation system for periodontal ligament stem cells (PDLSCs) to recreate the biological microenvironment similar to those of a regenerative milieu. Monodispersed human PDLSCs were cultured in medium with ascorbic acid and conditioned medium from developing apical tooth germ cells and were subsequently harvested from culture plate as a contiguous cell sheet with abundant extracellular matrix. The detached cell-matrix membrane spontaneously contracted to produce a single-cell pellet. The PDLSCs embedded within this cell-matrix complex exhibited several phenotypic characteristics of cementoblast lineages, as indicated by upregulated alkaline phosphatase activity, accelerated mineralization, and the expression of bone sialoprotein and osteocalcin genes. When this PDLSC pellets were transplanted into immunocompromised mice, a regular aligned cementum/PDL-like complex was formed. These results suggest that the combination of apical tooth germ cell-conditioned medium and endogenous extracellular matrix could maximally mimic the microenvironment of root/periodontal tissue development and enhance the reconstruction of physiological architecture of a cementum/PDL-like complex in a tissue-mimicking way; on the other hand, such PDLSC pellet may also be a promising alternative to promote periodontal defect repair for future clinical applications.
Yang
Zhenhua
Z
Department of Orthodontics, School of Stomatology, Fourth Military Medical University , Xi'an, People's Republic of China.
Jin
Fang
F
Zhang
Xiaojun
X
Ma
Dandan
D
Han
Chun
C
Huo
Na
N
Wang
Yinxiong
Y
Zhang
Yunfei
Y
Lin
Zhu
Z
Jin
Yan
Y
D000818:Animals; D001842:Bone and Bones / Q000187:drug effects; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation; D002516:Ceramics / Q000494:pharmacology; D003739:Dental Cementum / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D003804:Dentin / Q000187:drug effects / Q000502:physiology; D006801:Humans; D051379:Mice; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D023822:Tissue Engineering / Q000379:methods*; D014083:Tooth Germ / Q000166:cytology / Q000187:drug effects