COBL9 and COBL7 synergistically regulate root hair tip growth via controlling apical cellulose deposition.

Root hairs are cylindrical extensions of root epidermal cells that are important for the acquisition of water and minerals, interactions between plant and microbes. The deposition of cell wall materials in the tip enables root hairs to maintain elongation constantly. To date, our knowledge of the regulators that connect the architecture of cell wall and the root hair development remains very limited. Here, we demonstrated that COBL9 and COBL7, two genes of COBRA-Like family in Arabidopsis as well as their counterparts in rice, OsBC1L1 and OsBC1L8, regulate root hair growth. Single mutant cobl9, double mutants cobl7 cobl9 and double mutants osbc1l1 osbc1l8 all displayed prematurely terminated root hair elongation, though at varying levels. COBL7-YFP and COBL9-YFP accumulate prominently in the growing tips of newly emerged root hairs. Furthermore, cobl9, cobl7 cobl9 and osbc1l1 osbc1l8 mutants were defective in the enrichment of cellulose in the tips of the growing root hairs. We also discovered that overexpression of COBL9 could promote root hair elongation and salinity tolerance. Taken together, these results provide compelling evidence that the polarized COBL7 and COBL9 in the tip of the emerging root hairs have conserved roles in regulating root hair development and stress adaptation in dicots and monocots.

Accuracy of computer-aided prediction in soft tissue changes after orthodontic treatment.

INTRODUCTION: An accurate prediction in the soft tissue changes is of great importance for orthodontic treatment planning. Previous studies on the accuracy of the Dolphin visual treatment objective (VTO) in predicting treatment results were mainly focused on orthognathic treatment. The accuracy of Dolphin VTO prediction for orthodontic treatment is, however, poorly understood. The aim of this study was to evaluate the accuracy of Dolphin VTO prediction in soft tissue changes after orthodontic treatment by comparing the changes between predicted and actual values. METHODS: A total of 157 patients were screened for eligibility, and 34 young adult patients (8 males, 26 females; mean age 24.8 ± 3.9 years) were finally included in the study based on the inclusion and exclusion criteria. The landmarks and parameters of the Holdaway soft tissue analysis were used for the cephalometric analyses. The cephalometric tracings of the actual treatment result and the Dolphin predicted treatment outcome were superimposed to calculate the prediction errors. Paired t test was used to compare the statistical differences between the predicted and actual treatment outcomes of the parameters used in the Holdaway soft tissue analysis. RESULTS: There were significant differences between the predicted and actual values in parameters of the Holdaway soft tissue analysis (P < 0.05). The prediction of the landmarks in the lips region (ie, subnasale, soft tissue A-point, upper lip, lower lip, and soft tissue B-point) was inclined to be overestimated horizontally and underestimated vertically, whereas the prediction of the landmarks belonging to the chin region (ie, soft tissue pogonion, soft tissue gnathion, and soft tissue menton) was inclined to be underestimated horizontally and overestimated vertically. The most accurate prediction was found in the soft tissue A-point, whereas the least accurate one was found in the soft tissue in the chin region. The prediction was relatively more accurate in the vertical direction than in the horizontal direction. CONCLUSIONS: The Dolphin VTO prediction in soft tissue changes after the orthodontic treatment in patients with bimaxillary protrusion is the most accurate for the soft tissue A-point and the least accurate for the soft tissue chin region.

Thermosensitive progesterone hydrogel: a safe and effective new formulation for vaginal application.

PURPOSE: The safe and functional delivery of progesterone through the vaginal route remains an unmet clinical need. The purpose of this work is to prepare a new progesterone (P4) gel for vaginal application using a thermosensitive mucoadhesive polymer, glycol chitin (GC). METHOD: Thermogelling, mucoadhesive, mechanical, and viscoelastic properties of GC and the new formulation were evaluated using rheometry. In vitro release profile and the bioactivity of P4 were determined using vaginal fluid simulant (VFS) pH 4.2, and PR-reporter gene assay, respectively. In vitro safety of the formulations was tested using (VK2/E6E7) vaginal epithelial cell line and Lactobacillus Crispatus. Finally, in vivo safety and the efficacy of this formulation were evaluated using an endometrial hypoplasia mouse model. RESULTS: Results shows the aqueous solution of 5%; (w/v) GC loaded with 0.1%; (w/v) P4 prepared in pH 4.2, (GC-P4), forms a thermosensitive mucoadhesive hydrogel and can maintain stable physical properties at 37 °C. GC-P4 gel release 50% of P4 in 4 h after exposure to VFS, and no significant decrease in % viability of VK2/E6E7 or Lactobacillus was found after exposure to 5% GC or GC-P4. GC-P4 does not exhibit obvious toxicities to vaginal tissue in vivo even after repeated application. Efficacy studies indicated that GC-P4 was capable of preventing the progression of simple endometrial hyperplasia (SEH) to complex atypical endometrial hyperplasia (CAEH) in vivo. CONCLUSIONS: Results indicates that GC-P4 retains many characteristics for an effective vaginal delivery system for P4. Therefore we believe that GC-P4 formulation is a promising alternative to current vaginal P4 formulation.

Sema3A secreted by sensory nerve induces bone formation under mechanical loads.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling. Here, we focused on the role of Semaphorin 3A (Sema3A), expressed by sensory nerves, in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement (OTM) model. Firstly, bone formation was activated after the 3rd day of OTM, coinciding with a decrease in sensory nerves and an increase in pain threshold. Sema3A, rather than nerve growth factor (NGF), highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM. Moreover, in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells (hPDLCs) within 24 hours. Furthermore, exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload. Mechanistically, Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway, maintaining mitochondrial dynamics as mitochondrial fusion. Therefore, Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation, both as a pain-sensitive analgesic and a positive regulator for bone formation.

Prevalence and severity of apical root resorption during orthodontic treatment with clear aligners and fixed appliances: a cone beam computed tomography study.

BACKGROUND: Fixed appliances have been the mainstream for orthodontic treatment, while clear aligners, such as Invisalign system, have become increasingly popular. The prevalence of apical root resorption (ARR) in patients with clear aligners is still controversial. The aim of this study was to investigate and compare the prevalence and severity of ARR in patients treated with clear aligners and fixed appliances using cone beam computed tomography (CBCT). MATERIALS AND METHODS: A total of 373 roots from 70 subjects, with similar baseline characteristics and the ABO discrepancy index scores (i.e., treatment difficulty), were included into two groups: the clear aligners group (Invisalign, Align Technology, California, USA) and fixed appliances group (Victory Series; 3 M Unitek, California, USA). Root length of each anterior tooth was measured on the CBCT images by two blinded investigators. The ARR on each tooth was calculated as the difference of root length before and after orthodontic treatment. Chi-square test and paired t test was used to compare the ARR between the two groups as well as before and after orthodontic treatments. RESULTS: Prevalence of ARR in the clear aligners group (56.30%) was significantly lower than that in the fixed appliances group (82.11%) (P < 0.001). The severity of ARR in the clear aligners group (0.13 ± 0.47 mm) was significantly less than that in the fixed appliances group (1.12 ± 1.34 mm) (P < 0.001). The most severe ARR was found on the maxillary canine (1.53 ± 1.92 mm) and lateral incisor (1.31 ± 1.33 mm) in the fixed appliances group; the least ARR was found on the mandibular canine (- 0.06 ± 0.47 mm) and lateral incisor (0.04 ± 0.48 mm) in the clear aligners group (P < 0.001). CONCLUSIONS: The prevalence and severity of ARR measured on CBCT in patients with clear aligners were less than those in patients with fixed appliances.

Synthesis and characterization of a new photo-crosslinkable glycol chitosan thermogel for biomedical applications.

The major limitations of typical thermogelling polymers for practical applications are low gel stability and weak mechanical properties under physiological conditions. In this study, we have synthesized a new polysaccharide-based thermogelling polymer that can be photo-crosslinked by UV irradiation to form a mechanically resilient and elastic hydrogel. Methacrylated hexanoyl glycol chitosan (M-HGC), was synthesized by a series of chemical modifications, N-hexanoylation and N-methacrylation, of glycol chitosan (GC). Various M-HGC polymers with different methacryl group contents were synthesized and their thermogelling and photo-crosslinkable properties were evaluated. The M-HGCs demonstrated a thermo-reversible sol-gel transition behavior in aqueous solutions. The thermally-induced hydrogels could be chemically crosslinked by UV-triggered photo-crosslinking. From the cytotoxicity studies using MTT and the live/dead assay, the M-HGC hydrogels showed non-cytotoxicity. These photo-crosslinkable thermogelling M-HGC polymers may hold great promises for various biomedical applications, such as an injectable delivery system and 3D cell culture.

Glioma-targeting micelles for optical/magnetic resonance dual-mode imaging.

Surgical resection is the primary mode for glioma treatment, while gross total resection is difficult to achieve, due to the invasiveness of the gliomas. Meanwhile, the tumor-resection region is closely related to survival rate and life quality. Therefore, we developed optical/magnetic resonance imaging (MRI) bifunctional targeted micelles for glioma so as to delineate the glioma location before and during operation. The micelles were constructed through encapsulation of hydrophobic superparamagnetic iron oxide nanoparticles (SPIONs) with polyethylene glycol-block-polycaprolactone (PEG-b-PCL) by using a solvent-evaporation method, and modified with a near-infrared fluorescent probe, Cy5.5, in addition to the glioma-targeting ligand lactoferrin (Lf). Being encapsulated by PEG-b-PCL, the hydrophobic SPIONs dispersed well in phosphate-buffered saline over 4 weeks, and the relaxivity (r 2) of micelles was 215.4 mM(-1)·s(-1), with sustained satisfactory fluorescent imaging ability, which might have been due to the interval formed by PEG-b-PCL for avoiding the fluorescence quenching caused by SPIONs. The in vivo results indicated that the nanoparticles with Lf accumulated efficiently in glioma cells and prolonged the duration of hypointensity at the tumor site over 48 hours in the MR image compared to the nontarget group. Corresponding with the MRI results, the margin of the glioma was clearly demarcated in the fluorescence image, wherein the average fluorescence intensity of the tumor was about fourfold higher than that of normal brain tissue. Furthermore, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay results showed that the micelles were biocompatible at Fe concentrations of 0-100 µg/mL. In general, these optical/MRI bifunctional micelles can specifically target the glioma and provide guidance for surgical resection of the glioma before and during operation.

Glycol chitin-based thermoresponsive hydrogel scaffold supplemented with enamel matrix derivative promotes odontogenic differentiation of human dental pulp cells.

INTRODUCTION: Hydrogels have been widely studied as tissue engineering scaffolds over the past 2 decades because of their favorable biological properties. Recently, a new biodegradable glycol chitin-based thermoresponsive hydrogel scaffold (GC-TRS) was developed that can be easily applied as a mild viscous solution at room temperature but quickly transforms into a durable hydrogel under physiological conditions. The aim of this study was to investigate the effects of GC-TRS on the proliferation and odontogenic differentiation of colony-forming human dental pulp cells (hDPCs) in the presence of enamel matrix derivative. METHODS: Glycol chitin was synthesized by N-acetylation of glycol chitosan. The morphology of the thermoresponsive hydrogel scaffold was observed by using scanning electron microscopy. The sol gel phase transition of the aqueous solution of glycol chitin was investigated by using the tilting method and rheometer studies. hDPCs were isolated based on their ability to generate clonogenic adherent cell clusters. The effect of GC-TRS and collagen on cell viability was examined by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Expression of markers for odontogenic/osteogenic differentiation (ie, dentin sialophosphoprotein, dentin matrix protein-1, osteonectin, and osteopontin) was analyzed by performing real-time polymerase chain reaction. RESULTS: GC-TRS exhibited a highly macroporous and well-interconnected porous structure. The polymer solution existed in a mildly viscous sol state, but it transitioned to a gel state and did not flow above approximately 37°C. Rheometer studies showed that the glycol chitin solution exhibited a fast sol gel transition approximately at body temperature. GC-TRS and collagen did not inhibit cell viability until 7 days. Dentin sialophosphoprotein and dentin matrix protein-1 were expressed by cells cultured in GC-TRS at a higher level than that in cells cultured in collagen (P < .05). In both the scaffold groups, dentin sialophosphoprotein, dentin matrix protein-1, and osteopontin messenger RNA was up-regulated significantly in EMD-treated hDPCs when compared with the nontreated cells (P < .05). CONCLUSIONS: GC-TRS allowed the proliferation and odontogenic differentiation of hDPCs. Furthermore, the differentiation was facilitated by EMD. These results suggest that GC-TRS has the potential to be used in tissue engineering techniques for dentin regeneration.

Preparation and characterization of glycol chitin as a new thermogelling polymer for biomedical applications.

In this study, a new thermo-sensitive polymer, glycol chitin, was synthesized by controlled N-acetylation of glycol chitosan and evaluated as a thermogelling system. The physico-chemical properties of glycol chitins with different degrees of acetylation (DA) were investigated in terms of degradation, cytotoxicity, rheological properties, and in vitro and in vivo gel formation. Aqueous solutions of glycol chitins were flowable freely at room temperature but quickly became a durable gel at body temperature. Thermo-reversible sol-gel transition properties were observed with fast gelation kinetics. Glycol chitins with higher DA showed faster degradation in the presence of lysozyme. They exhibited no significant biological toxicity against human cell lines. An anti-cancer drug, doxorubicin, could be incorporated into the hydrogel by a simple mixing process and released in a sustained pattern over 13 days. Our findings suggest that glycol chitins could be useful as a new thermogelling biomaterial for drug delivery and injectable tissue engineering.