Uncoupling protein-2 regulates M1 macrophage infiltration of gingiva with periodontitis.

Periodontitis is an inflammatory disease accompanied by alveolar bone loss. Moreover, M1 macrophages play a critical role in the development of periodontal disease. Uncoupling protein-2 (UCP2) is a mitochondrial transporter protein that controls M1 macrophage activation by modulating reactive oxygen species (ROS) production. We investigated the role of UCP2 in M1 macrophage infiltration in gingival tissues with periodontitis. We found that the expression of UCP2 was upregulated in M1 macrophages infiltrating human periodontal tissues with periodontitis. Macrophage-specific knockout of UCP2 could increase the infiltration of macrophage and exacerbate inflammatory response in a mouse gingiva affected with periodontitis, induced by Porphyromonas gingivalis-LPS (Pg-LPS) injection. The loss of UCP2 may contribute to the enhanced abilities of proliferation, migration, pro-inflammatory cytokine secretion, and ROS production in Pg-LPS-treated macrophages. Our results indicate that UCP2 has an important role in M1 macrophage polarization in the periodontal tissue with periodontitis. It might be helpful to provide theoretical basis for design of new therapeutic strategies for periodontitis.

A comparative study of HAMSCs/HBMSCs transwell and mixed coculture systems.

Our previous studies indicated that a coculture system containing human amnion-derived mesenchymal stem cells (HAMSCs) and human bone marrow mesenchymal stem cells (HBMSCs) has the potential of application for bone regeneration. However, there is currently no enough comparative investigation between HAMSCs/HBMSCs transwell and mixed coculture systems. This study aimed to assess the phenotype and mechanisms regulated by indirect and direct coculture systems, respectively. Two in vitro models were employed with HAMSCs and HBMSCs at a ratio of 3:1, and then were analyzed by a series of processes, including flow cytometry, alkaline phosphatase (ALP) substrate assays, Alizarin red S staining, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and Western blot analysis. We found that cell proliferation, ALP activity, mineralized matrix formation, and osteoblast-related mRNA expression were accelerated in transwell coculture system compared with mixed coculture system. Conditioned medium from transwell coculture system achieved an elevated level of vascular endothelial growth factor and induced more vascular structures in human umbilical vein endothelial cells than those of mixed coculture system. Moreover, we observed that transwell coculture system, promoted osteogenesis and angiogenesis by maintaining stemness through extracellular regulated protein kinases 1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) signaling pathway. U0126, a selective inhibitor of ERK1/2 MAPK signaling, significantly suppressed maintaining of the stemness-based effects on transwell coculture system. Taken together, our results compared the merits of two different models and clarified the role of HAMSCs/HBMSCs transwell coculture system in the development of bone tissue engineering. © 2019 IUBMB Life, 2019.

Bioactive materials from berberine-treated human bone marrow mesenchymal stem cells promote alveolar bone regeneration by regulating macrophage polarization.

Alveolar bone regeneration has been strongly linked to macrophage polarization. M1 macrophages aggravate alveolar bone loss, whereas M2 macrophages reverse this process. Berberine (BBR), a natural alkaloid isolated and refined from Chinese medicinal plants, has shown therapeutic effects in treating metabolic disorders. In this study, we first discovered that culture supernatant (CS) collected from BBR-treated human bone marrow mesenchymal stem cells (HBMSCs) ameliorated periodontal alveolar bone loss. CS from the BBR-treated HBMSCs contained bioactive materials that suppressed the M1 polarization and induced the M2 polarization of macrophages in vivo and in vitro. To clarify the underlying mechanism, the bioactive materials were applied to different animal models. We discovered macrophage colony-stimulating factor (M-CSF), which regulates macrophage polarization and promotes bone formation, a key macromolecule in the CS. Injection of pure M-CSF attenuated experimental periodontal alveolar bone loss in rats. Colony-stimulating factor 1 receptor (CSF1R) inhibitor or anti-human M-CSF (M-CSF neutralizing antibody, Nab) abolished the therapeutic effects of the CS of BBR-treated HBMSCs. Moreover, AKT phosphorylation in macrophages was activated by the CS, and the AKT activator reversed the negative effect of the CSF1R inhibitor or Nab. These results suggest that the CS of BBR-treated HBMSCs modulates macrophage polarization via the M-CSF/AKT axis. Further studies also showed that CS of BBR-treated HBMSCs accelerated bone formation and M2 polarization in rat teeth extraction sockets. Overall, our findings established an essential role of BBR-treated HBMSCs CS and this might be the first report to show that the products of BBR-treated HBMSCs have active effects on alveolar bone regeneration.

An Injectable Black Phosphorus Hydrogel for Rapid Tooth Extraction Socket Healing.

The excess production of reactive oxygen species (ROS) will delay tooth extraction socket (TES) healing. In this study, we developed an injectable thermosensitive hydrogel (NBP@BP@CS) used to treat TES healing. The hydrogel formulation incorporated black phosphorus (BP) nanoflakes, recognized for their accelerated alveolar bone regeneration and ROS-scavenging properties, and dl-3-n-butylphthalide (NBP), a vasodilator aimed at enhancing angiogenesis. In vivo investigations strongly demonstrated that NBP@BP@CS improved TES healing due to antioxidation and promotion of alveolar bone regeneration by BP nanoflakes. The sustained release of NBP from the hydrogel promoted neovascularization and vascular remodeling. Our results demonstrated that the designed thermosensitive hydrogel provided great opportunity not only for ROS elimination but also for the promotion of osteogenesis and angiogenesis, reflecting the "three birds with one stone" concept, and has tremendous potential for rapid TES healing.

Injectable Thermosensitive Gel CH-BPNs-NBP for Effective Periodontitis Treatment through ROS-Scavenging and Jaw Vascular Unit Protection.

Periodontitis, a prevalent inflammatory condition in the oral cavity, is closely associated with oxidative stress-induced tissue damage mediated by excessive reactive oxygen species (ROS) production. The jaw vascular unit (JVU), encompassing both vascular and lymphatic vessels, plays a crucial role in maintaining tissue fluid homeostasis and contributes to the pathological process in inflammatory diseases of the jaw. This study presents a novel approach for treating periodontitis through the development of an injectable thermosensitive gel (CH-BPNs-NBP). The gel formulation incorporates black phosphorus nanosheets (BPNs), which are notable for their ROS-scavenging properties, and dl-3-n-butylphthalide (NBP), a vasodilator that promotes lymphatic vessel function within the JVU. These results demonstrate that the designed thermosensitive gel serve as a controlled release system, delivering BPNs and NBP to the site of inflammation. CH-BPNs-NBP not only protects macrophages and human lymphatic endothelial cells from ROS attack but also promotes M2 polarization and lymphatic function. In in vivo studies, this work observes a significant reduction in inflammation and tissue damage, accompanied by a notable promotion of alveolar bone regeneration. This research introduces a promising therapeutic strategy for periodontitis, leveraging the unique properties of BPNs and NBP within an injectable thermosensitive gel.

Morphometric analysis of paired fibula and mandible for optimal fibular mandibular reconstruction in a Chinese population.

To analyze the morphology of paired fibula and mandible aiming to choose optimal fibular segments for mandibular reconstruction in a Chinses population. A total of 118 cases of paired mandible and fibula was collected. All patients were received preoperative cone beam CT (CBCT) scans for mandibular evaluation and CT-angiographical (CTA) examination of the bilateral lower legs, respectively. The cross-sectional morphological differences between proximal (Side P), middle (Side M) and distal (Side D) segments of fibula and anterior, premolar and molar areas of mandible were compared. The most frequent cross-sectional shape at Side D, Side M and Side P portion of fibula was circular (75.4%), triangular (67.8%) and circular (49.2%), respectively. In anterior, premolar and molar areas of mandible, the most of the cross-section was s-shape (90.82%), straight (83.64%) and oblique (91.89%), respectively. The height and width of upper one third (W1) at Side M were significantly larger than those of Side D and Side P (p < 0.0001). There was significantly difference of width of lower one third (W2) among three groups (p < 0.0001). As for the height and widths of mandible, there was significant difference among anterior, premolar and molar regions (p < 0.0001). The rate of height between Side M of fibula and mandible (H (Side M/area)) was significantly larger than H (Side D/area) and H (Side P/area) (p < 0.01). The ratio of W1 between Side D of fibula and mandible (W1 (Side D/area)) was significantly larger than that of W1 (Side M/area) and W1 (side P/area) (p < 0.05). As for the ratio of W2 between fibula and mandible (W2 (plane/area)), there was significant difference among groups (p < 0.01). The distal and middle segments of fibula were suitable for reconstructing the anterior area of mandible and the proximal segment of fibula was more compatible with the premolar and molar areas of mandible.Clinical Relevance Presurgical morphometric analysis of paired fibula and mandible aids for optimal fibular-based mandibular reconstruction.

Polydopamine-Ag composite surface guides HBMSCs adhesion and proliferation.

Human bone marrow mesenchymal stem cells (HBMSCs) are regarded as an important resource in the field of maxillofacial bone regeneration because of their favorable properties when compared with other stem cells. Hence, finding suitable materials that could extend the application of HBMSCs has become an emerging medical topic and socioeconomic problem. In this work, polydopamine (PDA)-Ag surface was fabricated by PDA assisted photoreduction method, and the obtained PDA-Ag composite surface significantly promoted HBMSCs adhesion and proliferation. This effect is highly related to the amount of Ag nanoparticles (Ag NPs) present on the PDA surface. The behavior of HBMSCs on PDA-Ag surface could be spatially manipulated by controlling the distribution of Ag NPs on PDA surface (by controlling UV light). The general adhesion property allows the PDA-Ag surface to be fabricated on various substrates, making it a simple, general and controllable method for the fabrication of bioactive surface for HBMSCs.

Reconfigurable Surface with Photodefinable Physicochemical Properties for User-Designable Cell Scaffolds.

Surfaces with specific topography and chemical composition are quite useful in many applications ranging from functional interfaces to cell incubation scaffolds. Although these surfaces can be easily fabricated by combining topography-construction methods and surface-functionalization strategies, their properties are often static after fabrication or merely switchable between "on" and "off" states. Developing surfaces that can be on-demand regulated are quite important for the generation of smart surfaces for future applications. In this paper, we present a reconfigurable surface with adjustable topography and chemical functionality utilizing the photodynamic feature of the disulfide bond. Structured surfaces, composed of disulfide-cross-linked polymer networks, were prepared by using disulfide-containing methacrylate as the monomer. We show that the topography and chemical functionality of the surface can be on-demand regulated after its fabrication, with 254 and 365 nm UV light, respectively, allowing to "define" the physicochemical properties of the surface using light before the usage. We also demonstrate the application of such surface as a user-designable cell scaffold, that different cell scaffolds can be generated from one original surface with a simple exposure process, to define the desired bioactivity onto every point of the surface and therefore exactly control cell behaviors on the scaffold.

Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of human bone marrow mesenchymal stem cells via H19/miR-675/APC axis.

Bone volume inadequacy is an emerging clinical problem impairing the feasibility and longevity of dental implants. Human bone marrow mesenchymal stem cells (HBMSCs) have been widely used in bone remodeling and regeneration. This study examined the effect of long noncoding RNAs (lncRNAs)-H19 on the human amnion-derived mesenchymal stem cells (HAMSCs)-droved osteogenesis in HBMSCs. HAMSCs and HBMSCs were isolated from abandoned amniotic membrane samples and bone marrow. The coculture system was conducted using transwells, and H19 level was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). The mechanism was further verified. We here discovered that osteogenesis of HBMSCs was induced by HAMSCs, while H19 level in HAMSCs was increased during coculturing. H19 had no significant effect on the proliferative behaviors of HBMSCs, while its overexpression of H19 in HAMSCs led to the upregulated osteogenesis of HBMSCs in vivo and in vitro; whereas its knockdown reversed these effects. Mechanistically, H19 promoted miR-675 expression and contributed to the competitively bounding of miR-675 and Adenomatous polyposis coli (APC), thus significantly activating the Wnt/ß-catenin pathway. The results suggested that HAMSCs promote osteogenic differentiation of HBMSCs via H19/miR-675/APC pathway, and supply a potential target for the therapeutic treatment of bone-destructive diseases.

Development of hollow ferrogadolinium nanonetworks for dual-modal MRI guided cancer chemotherapy.

The development of hollow ferrogadolinium nanonetworks has not been reported for nanomedicine application until now. In this study, we developed a hollow and porous ferrogadolinium nanonetwork structure using the one-pot solvothermal method. This nanoparticle could be simultaneously used as a T 1 and T 2 dual-modal magnetic resonance imaging (MRI) contrast agent. In addition, the hollow lumen and abundant pores of the nanonetworks maximized the loading capacity and conferred the nanoplatforms for suitable anticancer drug loading capacity. Using these nanonetworks, MRI and anticancer experiments were conducted in vitro and satisfactory dual-modal MRI and cancer chemotherapy results were obtained. Therefore, the nanonetworks with dual-modal MRI and drug loading abilities effectively complement the ferrogadolinium composites' library and hold great promise in nanomedicine for simultaneous cancer diagnosis and chemotherapy.

Correction: Development of hollow ferrogadolinium nanonetworks for dual-modal MRI guided cancer chemotherapy.

[This corrects the article DOI: 10.1039/C8RA09102A.].

Composite of Gadolinium-Labeled Dendrimer Nanocluster And Graphene Oxide Nanosheet for Highly Efficient Liver T1-Weighted Imaging Probe.

Magnetic resonance contrast agent employs the use of gadolinium chelates, which has many limitations for clinical use including a low contrast effect, a short diagnostic window, and a brief blood circulation time. On this basis, we designed a gadolinium-labeled dendrimer nanocluster (GdDN) loaded graphene oxide nanosheet (GO-GdDN) to boost T1 contrast ability for imaging in vivo and improve blood circulation time. GO-GdDN presented an ultrahigh r1 relaxivity up to 19.07 mM-1 s-1 in a 9.4 T MR scanner, and a bright contrast image in vitro experiment. In addition, GO-GdDN could be internalized by HepG2 cells and presented strong cell contrast enhancement and reduction of the T1 value in HepG2 cells. In vivo, the retention time of GO-GdDN was significantly improved, so that the accumulation of GO-GdDN in liver was enhanced. Moreover, compared to Gd-DTPA, systemic delivery of GO-GdDN dramatically enhanced the signal-to-noise ratio of liver images, which was helpful for accurate imaging of liver and detection of liver lesions in vivo. Thus, this study demonstrates the utility of a powerful diagnosis tool for liver tumor or lesions.

LncRNA H19 Knockdown in Human Amniotic Mesenchymal Stem Cells Suppresses Angiogenesis by Associating with EZH2 and Activating Vasohibin-1.

Human amniotic mesenchymal stem cells (HAMSCs) are promising seed cells with great advantages in promoting angiogenesis. However, the mechanisms underlying angiogenesis facilitated by HAMSCs are still unclear. Long noncoding RNA H19 is involved in many biological processes, such as enhancing angiogenesis and proliferation, invasion, and migration of cancer cells. In this study, we constructed HAMSCs of stable low-expression H19 (HAMSC-shH19) and the scramble control (HAMSC-shNC) using lentiviral vectors, and in a three-dimensional coculture with human umbilical vein endothelial cells (HUVECs) to investigate the effect of H19 knockdown in HAMSCs on angiogenesis. Our results demonstrated that H19 knockdown significantly inhibited the angiogenic function of HAMSCs at an early stage in vitro and in vivo. The results of CCK8 and transwell assays demonstrated that the conditioned medium secreted by HAMSCs reduced proliferation and migration of HUVECs after downregulating H19. The angiogenesis factors expressed and secreted by HAMSC-shH19 were decreased compared with those secreted by the control, while angiogenesis inhibitors were elevated. Furthermore, we conducted chromatin immunoprecipitation and RNA-binding protein immunoprecipitation assays and found that H19 could interact with the histone methyltransferase Enhancer of Zeste homolog 2 (EZH2) and that H19 knockdown inhibited the ability of EZH2 to recruit methyl groups to the promoter region of the angiogenesis inhibitor gene vasohibin-1 (VASH1), thus increasing VASH1 expression and secretion of HAMSCs, suppressing angiogenesis. In summary, our study identified H19 as an important regulator in HAMSCs for promoting angiogenesis, which would help to construct ideal gene-modified seed cells to enhance angiogenesis in regenerative medicine.

Human amnion-derived mesenchymal stem cells promote osteogenesis of human bone marrow mesenchymal stem cells against glucolipotoxicity.

Epidemiological evidence suggests that diabetes mellitus (DM) is an important factor in promoting periodontitis. It not only affects the attachment of connective tissue but also causes loss of alveolar bone. Hence, there is an urgent need to find an effective treatment for DM-induced bone deficiency. This study aimed to investigate the effects of human amniotic mesenchymal stem cells (HAMSCs) on the proliferation and osteogenic differentiation of DM-induced human bone marrow mesenchymal stem cells (HBMSCs). High glucose and palmitic acid (GP) were used to mimic DM-induced glucolipotoxicity. The proliferation levels were measured using flow cytometry. Alkaline phosphatase activity substrate assays, Alizarin red S staining, and western blotting were used to investigate osteogenic differentiation. Oxidative stress was measured by assaying the levels of reactive oxygen species. This study found that glucolipotoxicity caused by GP remarkably inhibited cell proliferation and osteogenesis, and upregulated the oxidative stress level in HBMSCs. However, HAMSCs attenuated HBMSC dysfunction through antioxidant activity by influencing p38 mitogen-activated protein kinase and vascular endothelial growth factor secretion. In conclusion, our findings indicate that HAMSCs might be suitable for treating DM-mediated bone deficiency.

Fast Strategy to Functional Paper Surfaces.

Paper, with advantages of low-cost, easy fabrication and disposal, flexibility and renewability, is a suitable substrate material for various applications. Functionalization and patterning on paper substrates are commonly required in many applications. Although many methods have been developed to achieve this, they typically suffer from some drawbacks such as time-consuming process, specific device dependence, lack of flexibility, low patterning resolution, and so forth. Herein, we present a general and fast method to functionalize paper sheets and create patterns. The whole modification process can be completed in 10 min and can be applied on various types of paper substrates and other natural materials such as natural fabrics. By our method, many commonly used functional groups can be covalently attached and patterned on paper substrates, while the characteristic features of the original paper substrates, for example, color, transparency, and conductivity, can be perfectly retained after modification to allow these properties to be incorporated into the resultant materials. High-resolution patterns can be created on paper by applying a photomask during the modification or controlling the time of modification to precisely control the functionality at any area on the obtained paper substrates. We also show the potential applications of our method in the fabrication of superhydrophobic coatings and biomaterials.

The lateral cervical stria approach to selective neck dissection: a preliminary study

Background: This study aims to propose a lateral cervical stria approach for selective neck dissection (SND) in patients of early-stage oral malignancies. Material and methods: The lateral cervical stria approach was used in 11 patients undergoing SND between December 2020 and March 2022. The surgical incision was located in submandibular cervical stria, with a length of 5.0 cm. The ipsilateral SND was performed according to the pathological type, covering part or all of I-V levels. Perioperative variables including operation time, blood loss, drainage volume, number of lymph node as well as complications were assessed. The score of appearance using the University of Washington Quality of Life Questionnaire (UW-QOL) was recorded 6-month postoperatively. Results: Direct closure of primary lesion was performed in ten patients and a forearm free flap reconstruction was used in one patient. No wound breakdown or infection was found in all cases. The mean operative time of SND was 157.63±27.39 min. The volume of intraoperative blood loss and postoperative drainage was 120.45±36.77 ml and 314.09±98.82 ml, respectively. The mean number of retrieved lymph nodes was 17.89±6.03 (ranging from 12 to 31). Postoperative complications included mild static lower lip deviation (n=1), shoulder discomfort (n=1) and mild auricular paraesthesia (n=1). The mean score of appearance was 86.36±13.06, with 100 scores in 5 patients and 75 scores in 6 patients. Conclusions: The lateral cervical stria approach for SND in early-stage oral malignancies is reliable, achieving to satisfactory functional and aesthetic outcomes. (AU)