Age-related decline in the matrix contents and functional properties of human periodontal ligament stem cell sheets

Wu, Ruixue; Bi, Chun‐Sheng; Yu, Yang; Zhang, Lin-Lin; Chen, Fa‐Ming

doi:10.1016/j.actbio.2015.04.024

Cited by 58 publications

(72 citation statements)

References 70 publications

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“…The use of cell sheets can improve the survival rate of therapeutic cells and allows them to easily attach to host tissues. Moreover, the cell-matrix and cell-cell interactions in an intact sheet play an essential role in maintaining their cell genomic stability as 'building blocks' and their biochemical signaling and modulation of stem cell behavior toward therapeutic regeneration (Patel and Zhang 2014;Wu et al 2015). Most importantly, cell sheets make delivering and securing PDLSCs onto the tooth root surface possible, which is a key to PDL tissue regeneration (Dan et al 2014).…”

Section: Discussionmentioning

confidence: 99%

“…Briefly, the cells (P4) were seeded in 6-well culture dishes at a density of 2 × 10 5 cells per dish and pre-incubated for 24 h as described in the group design. Then, all the media were changed to a cell sheet-induction medium (complete medium supplemented with 50 μg/mL vitamin C) (Wu et al 2015) for cell sheet production. After 14 days, the resulting cell sheets were harvested using sterile tweezers and fixed in 4 % paraformaldehyde for 12 h. The obtained specimens were embedded in paraffin blocks and then cut into 5-μm-thick sections.…”

Section: Effects Of Different Pretreatments On Cell Sheet Productionmentioning

confidence: 99%

“…Then, the cell suspensions were washed twice, resuspended in PBS and analyzed using a Beckman Coulter Epics XL cytometer (Beckman Coulter, Fullerton, CA, USA). To further characterize the stem features of isolated PDLSCs, their proliferation ability was detected by the colony-forming assay and their differentiation potential was evaluated via osteogenic and adipogenic differentiation assays as described in our previous work (Wu et al 2015;Yu et al 2016).…”

Section: Identification Of Isolated Pdlscsmentioning

confidence: 99%

“…The experimental protocol was approved by the Institutional Review Board (IRB) of FMMU, School of Stomatology and an informed consent form agreeing to the contribution of their teeth for research purposes was signed by all donors. Human PDLSCs were isolated as previously described (Wu et al 2015). Briefly, the PDL tissue, which was scratched from the middle part of the root surface of the extracted teeth, was cut into small blocks and then digested for 1 h. After the tissue was digested, the digested tissue suspension was transferred to 6-well plates containing complete medium, i.e., α-minimum essential medium (α-MEM; Gibco BRL, Gaithersburg, MD, USA) supplemented with 10 % fetal bovine serum (FBS; Hangzhou Sijiqing Biological Engineering Materials, Zhejiang, China), 0.292 mg/mL glutamine (Invitrogen Life Technologies, Carlsbad, CA, USA), 100 U/mL penicillin (Gibco BRL) and 100 mg/mL streptomycin (Gibco BRL), for culturing the primary cells.…”

Section: Isolation and Culture Of Human Pdlscsmentioning

confidence: 99%

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Effects of short-term inflammatory and/or hypoxic pretreatments on periodontal ligament stem cells: in vitro and in vivo studies

et al. 2016

Cell Tissue Res

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In this study, we extensively screened the in vitro and in vivo effects of PDLSCs following short-term inflammatory and/or hypoxic pretreatments. We found that the 24-h hypoxic pretreatment of PDLSCs significantly enhanced cell migration and improved cell surface CXCR4 expression. In addition, hypoxia-pretreated PDLSCs exhibited improved cell colony formation and proliferation. Cells that were dually stimulated also formed more colonies compared to untreated cells but their proliferation did not increase. Importantly, the hypoxic pretreatment of PDLSCs enhanced cell differentiation as determined by elevated RUNX-2 and ALP protein expression. In this context, the inflammatory stimulus impaired cell OCN protein expression, while dual stimuli led to decreased RUNX-2 and OCN mRNA levels. Although preconditioning PDLSCs with inflammatory and/or hypoxic pretreatments resulted in no differences in the production of matrix proteins, hypoxic pretreatment led to the generation of thicker cell sheets; the inflammatory stimulus weakened the ability of cells to form sheets. All the resultant cell sheets exhibited clear bone regeneration following ectopic transplantation as well as in periodontal defect models; the amount of new bone formed by hypoxia-preconditioned cells was significantly greater than that formed by inflammatory stimulus- or dual-stimuli-treated cells or by nonpreconditioned cells. The regeneration of new cementum and periodontal ligaments was only identified in the hypoxia-stimulus and no-stimulus cell groups. Our findings suggest that PDLSCs that undergo short-term hypoxic pretreatment show improved cellular behavior in vitro and enhanced regenerative potential in vivo. The preconditioning of PDLSCs via combined treatments or an inflammatory stimulus requires further investigation.

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Section: Discussionmentioning

confidence: 99%

Section: Effects Of Different Pretreatments On Cell Sheet Productionmentioning

confidence: 99%

Section: Identification Of Isolated Pdlscsmentioning

confidence: 99%

Section: Isolation and Culture Of Human Pdlscsmentioning

confidence: 99%

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Effects of short-term inflammatory and/or hypoxic pretreatments on periodontal ligament stem cells: in vitro and in vivo studies

et al. 2016

Cell Tissue Res

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“…Periodontitis has a relatively high incidence; 46% of Americans exhibit chronic periodontitis (Eke et al, 2015), and the incidence is even higher in developing countries (Corbet, 2006). China exhibits a high incidence of periodontal disease; 80-97% of adults are affected (Wu et al, 2015). Periodontal health has therefore become a global health problem.…”

Section: Introductionmentioning

confidence: 99%

Hypoxia enhances periodontal ligament stem cell proliferation via the MAPK signaling pathway

He¹,

Cao²,

Zhang³

et al. 2016

Genet. Mol. Res.

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ABSTRACT. There is high incidence of periodontal disease in highaltitude environments; hypoxia may influence the proliferation and clone-forming ability of periodontal ligament stem cells (PDLSCs). The MAPK signaling pathway is closely correlated with cell proliferation, differentiation, and apoptosis. Thus, we isolated and cultured PDLSCs under hypoxic conditions to clarify the impact of hypoxia on PDLSC proliferation and the underlying mechanism. PDLSCs were separated and purified by the limiting dilution method and identified by flow cytometry. PDLSCs were cultured under hypoxic or normoxic conditions to observe their cloning efficiency. PDLSC proliferation at different oxygen concentrations was evaluated by MTT assay. Expression of p38/MAPK and MAPK/ERK signaling pathway members was detected by western blotting. Inhibitors for p38/MAPK or ERK were applied to PDLSCs to observe their impacts on clone formation and proliferation. Isolated PDLSCs exhibited typical stem cell morphological characteristics, strong abilities of globular clone formation and proliferation, and upregulated expression of mesenchymal stem cell markers. Stem cell marker expression was not statistically different between PDLSCs cultured under hypoxia and normoxia (P > 0.05). The clone number in the hypoxia group was significantly higher than that in the control (P < 0.05). PDLSC proliferation under hypoxia was higher than that of the control (P < 0.001). p38 and ERK1/2 phosphorylation in hypoxic PDLSCs was markedly enhanced compared to that in the control (P < 0.05). Either P38/MAPK inhibitor or ERK inhibitor treatment reduced clone formation and proliferation. Therefore, hypoxia enhanced PDLSC clone formation and proliferation by activating the p38/MAPK and ERK/MAPK signaling pathways.

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Analysis of senescence in gingival tissues and gingival fibroblast cultures

Furukawa

Matsuda

Aoki

et al. 2022

Clinical & Exp Dental Res

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Objective To determine senescence‐associated changes in the gingival tissues of aged mice and gingival fibroblast cultures. Materials and Methods The production of senescence‐associated β‐galactosidase (SA‐β‐gal) and mRNA expression of p16, p21, interleukin (IL)‐1β, and tumor necrosis factor α (TNF‐α) were evaluated in gingival tissues, gingival fibroblasts of 10‐ and 20‐month‐old C57BL/6NCrl mice, and multiple‐passaged and hydrogen peroxide‐stimulated human gingival fibroblasts (HGFs). Changes in molecular expression in HGF cultures due to senescent cell elimination by the senolytic drug ABT‐263 (Navitoclax) were analyzed. Results Compared to 10‐week‐old mice, the 20‐month‐old mice had higher numbers of M1 macrophages. The proportion of cells expressing SA‐β‐gal were also higher in 20‐ month‐old mice than in 10‐week‐old‐mice. Gingival fibroblasts in 20‐month‐old mice expressed less collagen 1a1, collagen 4a1, and collagen 4a2 mRNA than those in 10‐week‐old mice. Compared to control cells, H2O2 treated HGF cells expressed higher levels of SA‐β‐gal and p16, p21, IL‐1β, and TNF‐α. Furthermore, ABT‐263 suppressed HGF cell expression of cytokines after senescence induction. Conclusions Senescence‐associated changes were observed in the gingival tissues of aged mice and HGF cultures. In addition, the potential of senolytic drugs to modify aging‐related changes in the gingiva was shown.

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Age-related decline in the matrix contents and functional properties of human periodontal ligament stem cell sheets

Cited by 58 publications

References 70 publications

Effects of short-term inflammatory and/or hypoxic pretreatments on periodontal ligament stem cells: in vitro and in vivo studies

Effects of short-term inflammatory and/or hypoxic pretreatments on periodontal ligament stem cells: in vitro and in vivo studies

Hypoxia enhances periodontal ligament stem cell proliferation via the MAPK signaling pathway

Analysis of senescence in gingival tissues and gingival fibroblast cultures

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