Exosome-targeted delivery of METTL14 regulates NFATc1 m6A methylation levels to correct osteoclast-induced bone resorption.

Osteoporosis has a profound influence on public health. First-line bisphosphonates often cause osteonecrosis of the jaw meanwhile inhibiting osteoclasts. Therefore, it is important to develop effective treatments. The results of this study showed that the increased level of NFATc1 m6A methylation caused by zoledronic acid (ZOL), with 4249A as the functional site, is highly correlated with the decreased bone resorption of osteoclasts. Upstream, METTL14 regulates osteoclast bone absorption through the methylation functional site of NFATc1. Downstream, YTHDF1 and YTHDF2 show antagonistic effects on the post-transcriptional regulation of NFATc1 after the m6A methylation level is elevated by METTL14. In this study, meRIP-Seq, luciferase reporter assays, meRIP and other methods were used to elucidate the NFATc1 regulatory mechanism of osteoclasts from the perspective of RNA methylation. In addition, EphA2 overexpression on exosomes is an effective biological method for targeted delivery of METTL14 into osteoclasts. Importantly, this study shows that METTL14 released by exosomes can increase the m6A methylation level of NFATc1 to inhibit osteoclasts, help postmenopausal osteoporosis patients preserve bone mass, and avoid triggering osteonecrosis of the jaw, thus becoming a new bioactive molecule for the treatment of osteoporosis.

Polymeric coating on ß-TCP scaffolds provides immobilization of small extracellular vesicles with surface-functionalization and ZEB1-Loading for bone defect repair in diabetes mellitus.

Repair of critical-size bone defects in patients with diabetes mellitus (DM) has always been a challenge in clinical treatment. The process of bone defect regeneration can be impaired by underlying diseases including DM, but the mechanism remains unclear. In bone tissue engineering, the integration of bionic coatings and bioactive components into basic scaffolds are common function-enhancing strategies. Small extracellular vesicles (sEVs) have been applied for cell-free tissue regeneration in the last few years. We previously reported that sEVs have flexible and easily-extensible potential, through modular design and engineering modification. The impairment of CD31hiendomucinhi endothelial cells (ECs) whose function is coupling of osteogenesis and angiogenesis, is considered an important contributor to diabetic bone osteopathy, and ZEB1, which is highly expressed in CD31hiendomucinhi ECs, promotes angiogenesis-dependent bone formation. Thus we believe these ECs hold much promise for use in bone regeneration. In addition, c(RGDfC) has been reported to be a highly-effective peptide targeting αvß3, which is highly expressed in the bone microenvironment. In this study, we developed a hyaluronic acid (HA)/poly-L-lysine (PLL) layer-by-layer (LbL) self-assembly coating on ß-TCP (ß-tricalcium phosphate) scaffolds providing immobilization of modularized engineered sEVs (with c(RGDfC) surface functionalization and ZEB1 loading) to facilitate bone defect regeneration under DM conditions. RNA-seq was used to explore possible molecular mechanisms, and the therapeutic effects of bone regeneration were systematically evaluated in vitro and in vivo. Our data demonstrated that this strategy could be very effective in promoting the repair of diabetic bone defects, by enhancing angiogenesis, promoting osteogenesis and inhibiting osteoclast formation.

The biomechanical role of periodontal ligament in bonded and replanted vertically fractured teeth under cyclic biting forces.

After teeth are replanted, there are two possible healing responses: periodontal ligament healing or ankylosis with subsequent replacement resorption. The purpose of this study was to compare the fatigue resistance of vertically fractured teeth after bonding the fragments under conditions simulating both healing modes. Thirty-two human premolars were vertically fractured and the fragments were bonded together with Super-Bond C&B. They were then randomly distributed into four groups (BP, CP, CA, BA). The BP and CP groups were used to investigate the periodontal ligament healing mode whilst the BA and CA groups simulated ankylosis. All teeth had root canal treatment performed. Metal crowns were constructed for the CP and CA groups. The BP and BA groups only had composite resin restorations in the access cavities. All specimens were subjected to a 260 N load at 4 Hz until failure of the bond or until 2 × 106 cycles had been reached if no fracture occurred. Cracks were detected by stereomicroscope imaging and also assessed via dye penetration tests. Finally, interfaces of the resin luting agent were examined by scanning electron microscope. The results confirmed that the fatigue resistance was higher in the groups with simulated periodontal ligament healing. Periodontal reattachment showed important biomechanical role in bonded and replanted vertically fractured teeth.

[Effect of curing modes on knoop hardness and microtensile bond strength of two dentin adhesives].

OBJECTIVE: To evaluated the effect of curing modes and light-cure times on knoop hardness (KH) and microtensile bond strength (µTBS) of dentin adhesives in vitro. METHODS: Twenty molars were made into 80 dentin slices (about 1 mm thick). The dentin slices were prepared with an etch&rinse adhesive A (ONE-STEP PLUS) and a self-etch adhesive B (Clearfil SE Bond), and light-cured respectively under fast mode, i.e.1250 mW/cm(2) light intensity for 10 s, 15 s, 20 s, and ramp mode (soft start curing mode), i.e.initial 0 mW/cm(2) gradually increasing to 1250 mW/cm(2) in first 10 s, then steady for the next 10 s. The prepared dentin slices were kept in dark dry room for 24 h at 37°C, and KH were tested. The other 40 molars were flattened to expose coronal dentin, prepared with adhesives as above. Then the prepared teeth were restored with resin composites incrementally and cured under fast mode. The restored teeth were stored in water for 24 h at 37°C, and slowly sectioned to obtain multiple bonded beams. After 7 d water-storage, the samples received microtensile bond test, and the failure models of beams were observed under a stereomicroscope. Data were analyzed by ANOVA and LSD test (α = 0.05). RESULTS: No statistical difference in KH [(28.20 ± 5.36), (29.13 ± 5.60), (28.13 ± 4.40), (27.06 ± 3.77) MPa] and µTBS [(22.30 ± 5.07), (22.73 ± 6.59), (26.32 ± 6.17), (25.67 ± 4.31) MPa] of adhesive A were found between four curing conditions (fast mode for 10 s, 15 s, 20 s and ramp mode for 20 s) (P > 0.05). In adhesive B, KH of Fast 20 s [(28.23 ± 3.67) MPa] were significantly higher than those of Fast 10 s [(14.15 ± 2.24) MPa] and Fast 15 s [(17.63 ± 2.17) MPa] (P < 0.05). The µTBS of Fast 20 s [(42.52 ± 3.59) MPa] were significantly higher than those of Fast 10 s [(24.21 ± 3.60) MPa], Fast 15 s [(22.25 ± 4.16) MPa] and Ramp 20 s [(31.12 ± 5.40) MPa] (P < 0.05). In Fast 20 s and Ramp 20 s modes, there were no statistical difference in KH of adhesive A and B, while µTBS of adhesive B were higher than that of adhesive A(P < 0.05). CONCLUSIONS: As for different type dentin adhesives, the appropriate curing time in fast mode is different, and ramp mode (soft start curing mode) has no advantage over fast mode.