LIPUS promotes FOXO1 accumulation by downregulating miR-182 to enhance osteogenic differentiation in hPDLCs

Chen, Duanjing; Xiang, Mingli; Yang, Gong; Xu, Ling; Zhang, Tingwei; He, Yao; Zhou, Menglong; Xin, Liangjing; Li, Jie; Song, Jinlin

doi:10.1016/j.biochi.2019.08.005

Cited by 19 publications

(19 citation statements)

References 39 publications

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“…This could be because ALP is a marker of early osteogenic differentiation, and alizarin red staining may be more obvious in the later stage of osteoinduction. These results are similar to the previous cell osteogenesis studies, 27 suggesting that LIPUS could promote osteogenic differentiation of hPDLSCSs in vitro .…”

Section: Discussionsupporting

confidence: 92%

“…In this research, H&E staining results revealed that LIPUS‐treated hPDLSCSs are rich in cells and abundant ECM components, which may be one of the important reasons for promoting tissue regeneration. However, in our previous research, LIPUS did not significantly promote the proliferation of PDLCs 27 . A possible explanation is that LIPUS promotes further accumulation of ECM in cell sheets, which in turn regulates cell proliferation.…”

Section: Discussionmentioning

confidence: 56%

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Low‐intensity pulsed ultrasound promotes the formation of periodontal ligament stem cell sheets and ectopic periodontal tissue regeneration

Han

Zhou

Zhu

et al. 2020

J Biomedical Materials Res

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Human periodontal ligament stem cells (hPDLSCs) sheets play an important role in periodontal tissue engineering. Low‐intensity pulsed ultrasound (LIPUS) has been reported as an effective stimulus to regulate cell biological behavior. The present study aims to explore the potential of LIPUS to promote the formation and function of hPDLSC sheets (hPDLSCSs). Hematoxylin–eosin (H&E) staining, western blot, real‐time PCR, alkaline phosphatase (ALP), and alizarin red staining were used to evaluate the formation and osteogenic effect of LIPUS on hPDLSCSs in vitro. Hydroxyapatite with or without hPDLSCSs was transplanted in the subcutaneous pockets on the back of nude mice and histological analysis was performed. H&E staining showed increased synthesis of extracellular matrix (ECM) and real‐time PCR detected a significant increase in ECM‐related genes after LIPUS treatment. In addition, LIPUS could promote the expression of osteogenic differentiation‐related genes and proteins. ALP and alizarin red staining also found LIPUS enhanced the osteogenesis of hPDLSCSs. After transplantation in vivo, more dense collagen fibers similar to periodontal ligament were regenerated. Collectively, these results indicate that LIPUS not only promotes the formation and osteogenic differentiation of hPDLSCSs but also is a potential treatment strategy for periodontal tissue engineering.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 56%

Low‐intensity pulsed ultrasound promotes the formation of periodontal ligament stem cell sheets and ectopic periodontal tissue regeneration

Han

Zhou

Zhu

et al. 2020

J Biomedical Materials Res

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“…implied that LIPUS treatment effectively impaired osteoclast differentiation and produced a reduction in osteoclast markers. Most importantly, our previous studies have shown that LIPUS exerts potential protective effects on alveolar bone regeneration in periodontal injury models 11 and facilitates osteogenic differentiation of PDLCs 12 . These results were confirmed by micro-CT imaging, CEJ-ABC distances, BV/TV, Tb.Th., and H&E staining, which revealed that LIPUS significantly attenuated ligature-induced alveolar bone loss (Figure 1 ).…”

Section: Discussionmentioning

confidence: 96%

“…Multiple experiments have demonstrated that LIPUS modulates bone tissue regeneration and accelerates bone repair processes by upregulating bone specific genes and downregulating osteoclast differentiation 9 , 10 . Our latest studies indicated that LIPUS promoted alveolar bone regeneration in vivo 11 and facilitated osteogenic differentiation in vitro 12 . However, the precise mechanism by which LIPUS exerts protective effects on periodontitis remains unclear, thereby limiting the clinical application of LIPUS.…”

Section: Introductionmentioning

confidence: 99%

Low-intensity Pulsed Ultrasound regulates alveolar bone homeostasis in experimental Periodontitis by diminishing Oxidative Stress

Ying¹,

Tan²,

Ge³

et al. 2020

Theranostics

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Periodontitis is a widespread oral disease that results in the loss of alveolar bone. Low-intensity pulsed ultrasound (LIPUS), which is a new therapeutic option, promotes alveolar bone regeneration in periodontal bone injury models. This study investigated the protective effect of LIPUS on oxidative stress in periodontitis and the mechanism underlying this process. Methods: An experimental periodontitis model was induced by administering a ligature . Immunohistochemistry was performed to detect the expression levels of oxidative stress, osteogenic, and osteoclastogenic markers in vivo . Cell viability and osteogenic differentiation were analyzed using the Cell Counting Kit-8, alkaline phosphatase, and Alizarin Red staining assays. A reactive oxygen species assay kit, lipid peroxidation MDA assay kit, and western blotting were used to determine oxidative stress status in vitro . To verify the role of nuclear factor erythroid 2-related factor 2 (Nrf2), an oxidative regulator, during LIPUS treatment, the siRNA technique and Nrf2 -/- mice were used. The PI3K/Akt inhibitor LY294002 was utilized to identify the effects of the PI3K-Akt/Nrf2 signaling pathway. Results: Alveolar bone resorption, which was experimentally induced by periodontitis in vivo , was alleviated by LIPUS via activation of Nrf2. Oxidative stress, induced via H 2 O 2 treatment in vitro , inhibited cell viability and suppressed osteogenic differentiation. These effects were also alleviated by LIPUS treatment via Nrf2 activation. Nrf2 silencing blocked the antioxidant effect of LIPUS by diminishing heme oxygenase-1 expression. Nrf2 -/- mice were susceptible to ligature-induced periodontitis, and the protective effect of LIPUS on alveolar bone dysfunction was weaker in these mice. Activation of Nrf2 by LIPUS was accompanied by activation of the PI3K/Akt pathway. The oxidative defense function of LIPUS was inhibited by exposure to LY294002 in vitro . Conclusions: These results demonstrated that LIPUS regulates alveolar bone homeostasis in periodontitis by attenuating oxidative stress via the regulation of PI3K-Akt/Nrf2 signaling. Thus, Nrf2 plays a pivotal role in the protective effect exerted by LIPUS against ligature-induced experimental periodontitis.

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“…Since healthy PDLSCs are critical for maintaining tissue homeostasis, repair, and remodeling [28], many methods have been used to improve the differentiation ability of PDLSCs. Our previous studies have demonstrated that LIPUS causes increased H-PDLC ALP staining and Runx2 expression and matrix mineralization [16,29], and it also enhances new bone formation and periodontal regeneration in beagle dog models [30]. However, the effect of LIPUS on P-PDLSCs was unclear, which has limited the application of LIPUS in the treatment of periodontitis, because the inflammatory environment is unavoidable during periodontitis treatment.…”

Section: Discussionmentioning

confidence: 99%

Low-intensity pulsed ultrasound upregulates osteogenesis under inflammatory conditions in periodontal ligament stem cells through unfolded protein response

Deng

Tan

et al. 2020

Stem Cell Res Ther

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Background: In periodontal tissue engineering, periodontal ligament stem cells derived from patients with periodontitis (P-PDLSCs) are among the most promising and accessible stem cells for repairing disrupted alveolar bone and other connective tissues around the teeth. However, the inflammatory environment influences the osteogenic differentiation ability of P-PDLSCs. We examined low-intensity pulsed ultrasound (LIPUS) in P-PDLSCs in vitro and in rats with experimental periodontitis to determine whether LIPUS can enhance the osteogenic differentiation of stem cells. Materials and methods: P-PDLSCs were harvested and isolated from the periodontal tissues around the teeth of periodontitis patients, and healthy PDLSCs (H-PDLSCs) were obtained from tissues around healthy teeth. After validation by flow cytometry analysis, the P-PDLSCs were cultured in osteogenic medium either pretreated with the endoplasmic reticulum stress (ERS) inhibitor 4-phenyl butyric acid (4-PBA) or not pretreated and then treated with or without LIPUS (90 mW/cm 2 , 1.5 MHz) for 30 min per day. Cell viability, ERS marker expression, and osteogenic potential were determined between the different treatment groups. LPS-induced H-PDLSCs were used to mimic the inflammatory environment. In addition, we established a model of experimental periodontitis in rats and used LIPUS and 4-PBA as treatment methods. Then, the maxillary bone was collected, and micro-CT and histology staining methods were used to detect the absorption of alveolar bone. Results: Our data showed that the P-PDLSCs derived from periodontitis tissues were in a more pronounced ERS state than were the H-PDLSCs, which resulted in the former being associated with increased inflammation and decreased osteogenic ability. LIPUS can alleviate ERS and inflammation while increasing the bone formation capacity of P-PDLSCs in vivo and in vitro.

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LIPUS promotes FOXO1 accumulation by downregulating miR-182 to enhance osteogenic differentiation in hPDLCs

Cited by 19 publications

References 39 publications

Low‐intensity pulsed ultrasound promotes the formation of periodontal ligament stem cell sheets and ectopic periodontal tissue regeneration

Low‐intensity pulsed ultrasound promotes the formation of periodontal ligament stem cell sheets and ectopic periodontal tissue regeneration

Low-intensity Pulsed Ultrasound regulates alveolar bone homeostasis in experimental Periodontitis by diminishing Oxidative Stress

Low-intensity pulsed ultrasound upregulates osteogenesis under inflammatory conditions in periodontal ligament stem cells through unfolded protein response

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