Concentrated growth factor combined with iRoot BP Plus promotes inflamed pulp repair: an in vitro and in vivo study.

BACKGROUND: Platelet concentrates combined with calcium silicate cements may promote reparative dentin formation. However, few studies have reported their effect on dental pulp inflammation. This study aimed to evaluate the effects of concentrated growth factor (CGF) combined with iRoot BP Plus on inflammatory human dental pulp stem cells (hDPSCs) in vitro and inflamed pulp in rats in vivo. METHODS: The proliferation of LPS-stimulated hDPSCs treated with 50% CGF with/without 25% iRoot BP Plus was evaluated using Cell Counting Kit-8 on days 1, 4 and 7. The expression of genes associated with inflammation on day 1 and differentiation on day 14 was analysed by real-time polymerase chain reaction. The exposed pulp of rat maxillary molars was injected with 10 mg/mL LPS and directly capped with CGF membrane with/without iRoot BP Plus extract for 1, 7 and 28 days. The teeth were subjected to histologic analyses and immunohistochemistry. RESULTS: The proliferation rates of the inflammatory hDPSCs after the combination treatment were significantly higher than those after the other treatments on days 4 and 7 (P < 0.05). IL-1ß, IL-6, and TNF-α levels were increased in inflammatory hDPSCs but decreased after treatment with CGF combined with iRoot BP Plus extract, whereas IL-4 and IL-10 showed the opposite expression patterns. Expression of the odontogenesis-related genes OCN, Runx2, and ALP was dramatically enhanced by combined treatment with CGF and iRoot BP Plus extract. In rat pulp, the average inflammation scores of the CGF and CGF-iRoot BP Plus groups significantly decreased in comparison with those of the LPS group (P < 0.05), and the CGF-iRoot BP Plus group had more reparative dentin than the CGF and BP groups. Immunohistochemical staining showed fewer M1 macrophages on day 1 and more M2 macrophages on day 7 in the CGF-iRoot BP Plus group than in the other groups. CONCLUSIONS: The combination of CGF and iRoot BP Plus showed a synergistic effect on anti-inflammatory potential and promoted greater pulp healing than CGF or iRoot BP Plus alone.

Selective and Tunable Galectin Binding of Glycopolymers Synthesized by a Generalizable Conjugation Method.

Glycopolymers, conjugates of synthetic polymers with pendant carbohydrates, are becoming increasingly important to probe the role of carbohydrates in cellular processes and for applications like biosensors and drug delivery. A library of glycopolymers bearing different sugar moieties was synthesized by grafting amino-functionalized sugars to poly(acrylic acid) via DMTMM coupling. Primary amines were introduced at the anomeric (C-1) position to a number of unprotected mono-, di-, and trisaccharides using ammonium carbamate and conjugated to poly(acrylic acid) of different molecular weights, synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. This approach provides a simple and efficient route for the preparation of glycopolymers that differ only in the identity or degree of substitution of the sugar moiety on the polymer. The binding parameters (ka, kd, and KD) of these new glycopolymers to galectins-1 and -3 were quantified using surface plasmon resonance. The galectins selectively bound only to lactose-containing polymers, and the binding affinity was dependent on the galectin type, degree of sugar substitution and the molecular weight of polymer chains. Binding to both galectin-1 and -3 increased with a higher degree of sugar substitution, and higher molecular weight of the polymer backbone, reaching KD values on the order of 10-11 M.

Megalin-targeting and ROS-responsive elamipretide-conjugated polymeric prodrug for treatment of acute kidney injury.

Acute kidney injury (AKI) is associated with both kidney function loss and increased mortality. In the pathological progression of ischemia-reperfusion-induced AKI, the surge of reactive oxygen species (ROS) plays a crucial role. To combat this, mitochondrial-targeted antioxidant therapy shows great promise as mitochondria are the primary source of ROS in AKI. However, most strategies aiming to target mitochondria directly result in nanodrugs that are too large to pass through the glomerular system and reach the renal tubules, which are the main site of damage in AKI. This study focused on synthesizing a Megalin receptor-targeted polymeric prodrug, low molecular weight chitosan-thioketal-elamipretide (LMWC/TK/Ela), to mitigate excessive ROS in renal tubular epithelial cells for AKI. This soluble polymeric prodrug has the ability to successfully reach the tubular site by crossing the glomerular barrier. Once there, it can responsively release elamipretide, which possesses excellent antioxidative properties. Therefore, this research offers a novel approach to actively target renal tubular epithelial cells and intracellular mitochondria for the relief of AKI.

Tough and elastic hydrogels based on robust hydrophobicity-assisted metal ion coordination for flexible wearable devices.

Flexible wearable sensors that combine excellent flexibility, high elasticity, sensing capabilities, and outstanding biocompatibility are gaining increasing attention. In this study, we successfully develop a robust and elastic hydrogel-based flexible wearable sensor by modulating molecular structures combined with metal ion coordination. We leverage three N-acryloyl amino acid monomers, including N-acryloyl glycine (AG), N-acryloyl alanine (AA), and N-acryloyl valine (AV) with different hydrophobic groups adjacent to the carboxyl group, to copolymerize with acrylamide (AM) in the presence of Zr4+ for hydrogel preparation in one step (P(AM3-AG/AA/AV0.06)-Zr0.034+ hydrogels). Our investigation reveals that the P(AM3-AV0.06)-Zr0.034+ hydrogel with the most hydrophobic side group demonstrates superior mechanical properties (1.1 MPa tensile stress, 3566 kJ m-3 toughness and 1.3 kJ m-2 fracture energy) and resilience to multiple tensile (30% strain, 500 cycles) and compression cycling (50% strain, 500 cycles). Moreover, the P(AM3-AV0.06)-Zr0.034+ hydrogel exhibits good biocompatibility and high conductivity (1.1 S m-1) and responsivity (GF = 16.21), and is proved to be suitable as a flexible wearable sensor for comprehensive human activity monitoring.

The pluripotent factor OCT4A enhances the self-renewal of human dental pulp stem cells by targeting lncRNA FTX in an LPS-induced inflammatory microenvironment.

BACKGROUND: Regulating the pluripotency of human dental pulp stem cells (hDPSCs) is key for the self-repair of injured dental pulp. We previously found that OCT4A promotes the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs). Recent studies have shown the interaction between OCT4A and lncRNAs in pluripotency maintenance of various stem cells. The aim of this study was to explore the underlying roles and mechanisms of OCT4A and its related lncRNAs in the proliferation and multidirectional differentiation of hDPSCs in an inflammatory microenvironment. METHODS: Human lncRNA microarrays were applied to screen out the differentially expressed lncRNAs in hDPSCs between the OCT4A-overexpressing and vector groups. Lipopolysaccharide (LPS) was used to simulate the inflammatory microenvironment. The effects of OCT4A and the lncRNA FTX on the proliferation and multidifferentiation of hDPSCs were observed by the CCK-8 assay, EdU staining, real-time PCR, western blotting, and Alizarin red and oil red O staining. Bioinformatics analysis and chromatin immunoprecipitation (ChIP) assays were performed to clarify the targeted mechanism of OCT4A on FTX. The regulation by FTX of the expression of OCT4A and its downstream pluripotent transcription factors SOX2 and c-MYC was further detected by real-time PCR and western blotting. RESULTS: The microarray results showed that 978 lncRNAs (250 of which were upregulated and 728 downregulated) were potentially differentially expressed genes (fold change ≥ 2, P < 0.05). LPS stimulation attenuated the self-renewal of hDPSCs. OCT4A enhanced the cell proliferation and multidifferentiation capacities of hDPSCs in an inflammatory microenvironment, while FTX exhibited the opposite effects. OCT4A negatively regulated FTX function by binding to specific regions on the FTX promoter, thereby inhibiting the transcription of FTX. Moreover, overexpression of FTX downregulated the expression of OCT4A, SOX2 and c-MYC, whereas knockdown of FTX facilitated their expression. CONCLUSIONS: OCT4A was found to be a crucial factor maintaining the self-renewal of hDPSCs by transcriptionally targeting FTX in an inflammatory microenvironment. Moreover, we proposed a novel function of FTX in negatively regulating the pluripotency and multilineage differentiation capacity of hDPSCs. The hierarchical organization between OCT4A and FTX expanded the understanding of the network between transcription factors and lncRNAs in fine-tuning the pluripotency/differentiation balance of adult stem cells, and provided prospective targets for optimizing dental-derived stem cell sources for regenerative endodontics.

Efficient formation of multicompartment hydrogels by stepwise self-assembly of thermoresponsive ABC triblock terpolymers.

The gelation behavior of a poly(ethylene-alt-propylene)-b-poly(ethylene oxide)-b-poly(N-isopropylacrylamide) (PON) triblock terpolymer and a poly(N-isopropylacrylamide)-b-poly(ethylene oxide)-b-poly(N-isopropylacrylamide) (NON) triblock copolymer was studied by rheology over the concentration range 1-5 wt %. In comparison to the NON copolymer, gelation of the PON terpolymer was achieved at a much lower concentration, with a much sharper sol-gel transition. This is due to a stepwise gelation of PON terpolymers involving micellization at room temperature and gelation at elevated temperatures. The separation of micellization and gelation leads to the formation of a two-compartment network as observed by cryoTEM. The results highlight the intricate and tunable nanostructures and new properties accessible from ABC terpolymer hydrogels.

ABC triblock terpolymers exhibiting both temperature- and pH-sensitive micellar aggregation and gelation in aqueous solution.

Two poly(ethylene-alt-propylene)-b-poly(ethylene oxide)-b-poly(N-isopropylacrylamide-co-acrylic acid) (PEP-PEO-P(NIPAm-co-AA)) triblock terpolymers were synthesized by a combination of anionic and RAFT polymerizations, followed by acid hydrolysis. Micellar aggregation and gelation behavior in aqueous solutions were studied by dynamic light scattering (DLS) and rheology, respectively. DLS measurements on dilute solutions revealed that the triblock terpolymers form micelles with PEP cores and PEO-P(NIPAm-co-AA) coronae at room temperature and undergo a micelle to micellar aggregate transition upon heating. Rheological measurements showed that micellar aggregation manifests itself as gelation at higher concentrations (~4 wt %). The observed thermoresponsive aggregation and gelation is due to the intermicellar association of P(NIPAm-co-AA) blocks in the coronae above the lower critical solution temperature of the P(NIPAm-co-AA) block. The critical micellar aggregation and gelation temperatures are controlled by the mole fraction and degree of acrylic acid (AA) ionization in the P(NIPAm-co-AA) block, and therefore they can be modulated as functions of both pH and AA content in the polymer.

Polydopamine constructed interfacial molecular bridge in nano-hydroxylapatite/polycaprolactone composite scaffold.

Nano-hydroxylapatite (nano-HAP)/polycaprolactone (PCL) composite scaffold is proved to possess great potential for bone tissue engineering application since the biocompatibility of PCL and the osteoinduction ability of nano-HAP. However, the interfacial bonding between nano-HAP and PCL is weak by reason of the difference in thermodynamic properties. Herein, nano-HAP was modified by polydopamine (PDA) and then added to the PCL matrix to enhance their interface bonding in bone scaffold manufactured by selective laser sintering (SLS). The results indicated that PDA acted as an interfacial molecular bridge between PCL and nano-HAP. On one hand, the amino groups of PDA formed hydrogen bonding with the hydroxyl groups of nano-HAP, and on the other hand, the catechol groups of PDA formed hydrogen bonding with the ester groups of PCL. Compared with the HAP/PCL scaffolds, the tensile and compressive strength of the P-HAP/PCL scaffolds loading 12 wt% P-HAP were increased by 10% and 16%, respectively. Meanwhile, the scaffold possessed great bioactivity and cytocompatibility that could accelerate the formation of apatite layers and promote the cell adhesion, proliferation and differentiation.

[Expression of Forkhead boxO1 and Runt-related transcription factor 2 in periodontal tissue during orthodontic movement of teeth in rats].

PURPOSE: To investigate the expression of FoxO1 and Runx2 in periodontal tissue and their effect during orthodontic teeth movement(OTM) in rats. METHODS: Forty 8-week old male Sprague-Dawley (SD) rats were used to establish animal models of orthodontic teeth movement and divided into 5 groups randomly. The right side of jaws of each rat was set as experimental side, and the left side as control side. At 1, 3, 5, 7, 14 d after orthodontic treatment, the rats were sacrificed and the maxillary bone containing the first molar was dissected. H-E staining and immunohistochemical staining were used to detect the morphological changes, the expression of FoxO1 and Runx2 of the periodontal tissue at different points. Computer image analysis was used to evaluate the expression of FoxO1 and Runx2 in the periodontal tissues of the rats. The differences were analyzed by using SPSS19.0 software package. RESULTS: The expression of FoxO1 in periodontal ligament was mainly in osteoblasts and cementoblasts; and the expression of Runx2 was mainly in osteoblasts, fibroblasts and cementoblasts. In the experimental group,the expressions of FoxO1 and Runx2 in the periodontal tissues of rats increased significantly, reached the peak within 3-5 days, then decreased. There was no significant difference between the experimental group and the control group at 14th day (P>0.05), but significant difference was found between other group and control group (P<0.05). CONCLUSIONS: FoxO1 and Runx2 play a role in the reconstruction of periodontal tissue during orthodontic movement of teeth, and they are mainly involved in the process of osteoblast formation and bone formation.

A Selective Joint Determination of Salicylic Acid in Actinidia chinensis Combining a Molecularly Imprinted Monolithic Column and a Graphene Oxide Modified Electrode.

A new combination between selective polymer monolith microextraction (PMME) and sensitive differential pulse voltammetry (DPV) was developed for the determination of the phytohormone salicylic acid (SA) in Actinidia chinensis. A molecularly imprinted monolithic column (MIMC) thermally in-situ polymerized in a micropipette tip by using SA as a template, 4-vinyl pyridine (4-VP) as a functional monomer and ethylene glycol dimethacrylate (EGDMA) as a cross-linker in the mixed porogen of toluene and dodecanol, was employed for the microextraction of SA. The prepared MIMC was characterized by a Fourier transform infrared spectrometer (FI-TR), scanning electron microscope (SEM) and thermo gravimetric analysis (TGA). The results confirmed the binary continuous structure of the porous network. The extracted SA was determined by DPV on a graphene oxide (GO) modified electrode. The joint conditions between MIMC and DPV were investigated practically. Under the optimum conditions, SA could be determined selectively and sensitively in a linear range from 0.1 to 60.0 µg g-1. The limit of detection was 0.03 µg g-1 and the recoveries were between 86.2 and 105.2%. The proposed joint method was successfully used to determine SA in Actinidia chinensis.

[Relationship between masseter muscle electromyographic activities and the position of the mandible].

OBJECTIVE: To investigate the changes in electromyographic activities of the temporal and masseter muscles at different positions of the mandible. METHODS: Twenty orthodontic patients with Angle Class II malocclusion and mandibular retrusion (ANB<6°) aged 10-14 years were enrolled in this study. All the patients were treated with Forsus fixed functional appliance combined with MBT straight-wire appliance. The electromyographic activities of the temporal (T) and masseter (M) muscles before, during and after functional treatment were evaluated by assessing the average integrated electromyogram (EMG) and T/M ratio at clenching status in different mandibular positions. RESULTS: After functional forward positioning of the mandible, the electromyographic activities of the temporal and masseter muscles decreased and T/M ratio increased significance at the clenching status (P<0.05). CONCLUSION: The appliance insertion and activation is associated with a decreased EMG activity of the temporal and masseter muscles, and the T/M ratio is correlated to the position of the mandible.