Lei Hu1,2, Bin Zhao1, Zhenhua Gao1, Junji Xu1, Zhipeng Fan3, Chunmei Zhang1, Jinsong Wang4, Songlin Wang1,4. 1. Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, Beijing, China. 2. Department of Prosthodontics, Capital Medical University School of Stomatology, Beijing, China. 3. Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, School of Stomatology, Capital Medical University, Beijing, China. 4. Department of Biochemistry and Molecular Biology, Capital Medical University School of Basic Medical Sciences, Beijing, China.
Abstract
BACKGROUND: Although cell sheets have gained much interest as a non-scaffold strategy for tissue regeneration, the regenerative features of different cell sheets remain unclear. OBJECTIVE: In this study, we aimed to compare the regeneration characteristics of cell sheets derived from dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs) and stem cells of the apical papilla (SCAPs). METHODS: Dental pulp stem cells, PDLSCs and SCAPs from the same individual were acquired and induced to form sheets using 20 μg/mL vitamin C. Immunofluorescence staining was used to detect the expression of collagen I, fibronectin, integrin β1 and vimentin. Real-time PCR was used to determine NANOG, OCT4, SOX2 and TERT gene expression. The cell sheets with hydroxyapatite/tricalcium phosphate were transplanted into nude mice subcutaneously to evaluate tissue regeneration characteristics. RESULTS: No obvious differences were found in the histological structure and extracellular matrix protein expression between DPSC, PDLSC and SCAP sheets. Dental pulp stem cell sheet showed higher expression of OCT4 and TERT than PDLSC and SCAP sheets. All three cell sheets displayed the ability of mineral tissue formation and highly expressed periostin. The tissue derived from DPSC sheet showed higher CD31 expression and porous fibres compared with that from the others. The tissue fibres formed from PDLSC sheet were directionally arranged, while the tissue derived from SCAP sheet showed highest mineral tissue formation. CONCLUSION: Although in vitro DPSC, PDLSC and SCAP cell sheets have similar characteristics, their regenerative characteristics in vivo are different, with each showing potential application for regeneration of different tissues.
BACKGROUND: Although cell sheets have gained much interest as a non-scaffold strategy for tissue regeneration, the regenerative features of different cell sheets remain unclear. OBJECTIVE: In this study, we aimed to compare the regeneration characteristics of cell sheets derived from dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs) and stem cells of the apical papilla (SCAPs). METHODS: Dental pulp stem cells, PDLSCs and SCAPs from the same individual were acquired and induced to form sheets using 20 μg/mL vitamin C. Immunofluorescence staining was used to detect the expression of collagen I, fibronectin, integrin β1 and vimentin. Real-time PCR was used to determine NANOG, OCT4, SOX2 and TERT gene expression. The cell sheets with hydroxyapatite/tricalcium phosphate were transplanted into nude mice subcutaneously to evaluate tissue regeneration characteristics. RESULTS: No obvious differences were found in the histological structure and extracellular matrix protein expression between DPSC, PDLSC and SCAP sheets. Dental pulp stem cell sheet showed higher expression of OCT4 and TERT than PDLSC and SCAP sheets. All three cell sheets displayed the ability of mineral tissue formation and highly expressed periostin. The tissue derived from DPSC sheet showed higher CD31 expression and porous fibres compared with that from the others. The tissue fibres formed from PDLSC sheet were directionally arranged, while the tissue derived from SCAP sheet showed highest mineral tissue formation. CONCLUSION: Although in vitro DPSC, PDLSC and SCAP cell sheets have similar characteristics, their regenerative characteristics in vivo are different, with each showing potential application for regeneration of different tissues.
Authors: Harim T Dos Santos; Kyungsook Kim; Teruo Okano; Jean M Camden; Gary A Weisman; Olga J Baker; Kihoon Nam Journal: Cells Date: 2020-12-09 Impact factor: 6.600