No abstract

Hesperetin (3′,5,7-trihydroxy-4-methoxyflavanone) is a metabolite of hesperidin (hesperetin-7-O-rutinoside), which belongs to the flavanone subgroup and is found mainly in citrus fruits. Hesperetin has been reported to be an effective osteoinductive compound in various in vivo and in vitro models. However, how hesperetin effects osteogenic differentiation is not fully understood. In this study, we investigated the capacity of hesperetin to stimulate the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) and to relieve the anti-osteogenic effect of high glucose. Osteogenesis of PDLSCs was assessed by measurement of alkaline phosphatase (ALP) activity, and evaluation of the mRNA expression of ALP, runt-related gene 2 (Runx2), osterix (OSX), and FRA1 as osteogenic transcription factors, as well as assessment of protein expression of osteopontin (OPN) and collagen type IA (COLIA). When PDLSCs were exposed to a high concentration (30 mM) of glucose, osteogenic activity decreased compared to control cells. Hesperetin significantly increased ALP activity at doses of 1, 10, and 100 µM. Pretreatment of cells with hesperetin alleviated the high-glucose-induced suppression of the osteogenic activity of PDLSCs. Hesperetin scavenged intracellular reactive oxygen species (ROS) produced under high glucose condition. Furthermore, hesperetin increased the activity of the PI3K/Akt and β-catenin pathways. Consistent with this, blockage of Akt or β-catenin diminished the protective effect of hesperetin against high glucose-inhibited osteogenic differentiation. Collectively, our results suggest that hesperetin alleviates the high glucose-mediated suppression of osteogenic differentiation in PDLSCs by regulating ROS levels and the PI3K/Akt and β-catenin signaling pathways.

Low-level laser irradiation can promote the healing process in soft and hard tissue but the precise mechanisms are unclear. In this study, we examined the effect of LLLT (low-level laser therapy) on the healing of extraction sockets in diabetic and healthy rats. Forty-eight Sprague-Dawley rats were divided into normal (n = 24) and diabetic (n = 24) rats, and streptozotocin (STZ) injection was used to induce diabetes in the latter. The left and right maxillary first molars of all the rats were extracted. In the non-diabetic rats, the left extraction sockets were not irradiated (group 1) and the right ones were irradiated daily for 3, 5, 7, and 14 days after extraction with a galium-aluminum-arsenide (GaAlAs) diode laser (group 2), and in the diabetic rats, similarly the left ones were not irradiated (group 3) and the right ones were irradiated (group 4). Specimens acquired at these intervals were examined by hematoxylin and eosin (H&E) staining and reverse transcription polymerase chain reaction (RT-PCR). Histological observations and gene expression analyses revealed that groups 2 (normal rats with LLLT) and 4 (diabetic rats with LLLT) showed faster initial healing and more new alveolar bone formation than group 1 (normal rats without LLLT) and group 3 (diabetic rats without LLLT), respectively. We conclude that 980-nm GaAlAs low-intensity diode laser irradiation is beneficial for the initial stages of alveolar bone healing and for further calcification in both diabetic and normal rats when applied every day at a dose of 13.95 J/cm(2) for 60 s.

Objectives:To evaluate the long-term influence of residual bone height (RBH), sinus membrane perforation and the presence of voids on the survival of implants placed simultaneously with lateral maxillary sinus floor augmentation (MSFA). Materials and Methods:A total of 207 patients (613 implants) who underwent MSFA from 1999 to 2003 and followed up until 2018 were included in this study. RBH and marginal bone loss were assessed using a series of panoramic and periapical radiographs. Cumulative survival rates (CSR) with log-rank tests and hazard ratios of implant failure according to RBH, membrane perforation, and the presence of voids were assessed. Results:The overall 10-year and 20-year CSRs were 95% (95% CI: 84.95%-85.05%) and 85% (95% CI: 84.95%-85.05%), respectively. CSR was significantly higher for implants with ≥3 mm RBH (n = 260, 92.4%) than those with <3 mm RBH (n = 353, 78.8%) (p = .002). CSR of non-smoking group (n = 312, 90.0%) was higher than that of smoking group (n = 301, 77.1%) (p = .009). There was no significant difference in the hazard ratio of implant failure between the perforated membrane group (n = 245) and the non-perforated group (n = 368). In two out of 11 patients with voids, the inflammation of peri-implantitis progressed into voids, resulting in severe bone resorption. Conclusions:The survival of implants placed in <3 mm of RBH, even though it showed a lower CSR, could be considered acceptable. Neither sinus membrane perforation nor the presence of voids appeared to affect implant survival as long as supportive maintenance therapy with proper oral hygiene was provided. K E Y W O R D Sbone regeneration, bone substitutes, guided tissue regeneration, sinus floor elevation, smoking

Lee S‐I, Kang K‐L, Shin S‐I, Herr Y, Lee Y‐M, Kim E‐C. Endoplasmic reticulum stress modulates nicotine‐induced extracellular matrix degradation in human periodontal ligament cells. J Periodont Res 2012; 47: 299–308. © 2012 John Wiley & Sons A/S Background and Objective:  Tobacco smoking is considered to be one of the major risk factors for periodontitis. For example, about half the risk of periodontitis can be attributable to smoking in the USA. It is evident that smokers have greater bone loss, greater attachment loss and deeper periodontal pockets than nonsmoking patients. It has recently been reported that endoplasmic reticulum (ER) stress markers are upregulated in periodontitis patients; however, the direct effects of nicotine on ER stress in regard to extracellular matrix (ECM) degradation are unclear. The purpose of this study was to examine the effects of nicotine on cytotoxicity and expression of ER stress markers, selected ECM molecules and MMPs, and to identify the underlying mechanisms in human periodontal ligament cells. We also examined whether ER stress was responsible for the nicotine‐induced cytotoxicity and ECM degradation. Material and Methods:  Cytotoxicity and cell death were measured by 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5 diphenyltetrazolium bromide assay and flow cytometric annexin V and propidium iodide staining. The mRNA and protein expressions of MMPs and ER markers were examined by RT‐PCR and western blot analysis. Results:  Treatment with nicotine reduced cell viability and increased the proportion of annexin V‐negative, propidium iodide‐positive cells, an indication of cell death. Nicotine induced ER stress, as evidenced by survival molecules, such as phosphorylated protein kinase‐like ER‐resident kinase, phosphorylated eukaryotic initiation factor‐2α and glucose‐regulated protein‐78, and apoptotic molecules, such as CAAT/enhancer binding protein homologous protein (CHOP). Nicotine treatment led to the downregulation of ECM molecules, including collagen type I, elastin and fibronectin, and upregulation of MMPs (MMP‐1, MMP‐2, MMP‐8 and MMP‐9). Inhibition of ER stress by salubrinal and transfection of CHOP small interfering RNA attenuated the nicotine‐induced cell death, ECM degradation and production of MMPs. Salubrinal and CHOP small interfering RNA inhibited the effects of nicotine on the activation of Akt, JNK and nuclear factor‐κB. Conclusion:  These results indicate that nicotine‐induced cell death is mediated by the ER stress pathway, involving ECM degradation by MMPs, in human periodontal ligament cells.