Traumatic occlusion aggravates bone loss during periodontitis and activates Hippo-YAP pathway.

AIM: This study aimed at exploring changes in YAP expression and their effect on periodontitis (PD) combined with traumatic occlusion (TO). MATERIALS AND METHODS: BALB/cJ mice were used to establish a PD model by local administration of Porphyromonas gingivalis (P.g, ATCC 33277) and a TO model by occlusal elevation (OE) using composite resin bonding on the bilateral maxillary molar. The mouse fibroblast cell line (L929) and pre-osteoblast cell line (MC3T3-E1) were subjected to cyclic tensile/compressive stress and inflammatory stimuli (lipopolysaccharide from Escherichia coli) to verify in vivo results. RESULTS: Severe bone resorption was observed by microCT scanning in OE with P.g group, when compared to OE only and P.g only groups. Mechanical stress caused by OE activated the Hippo-YAP pathway in periodontal tissues and upregulated the expression of JNK/AP-1. OE with P.g further promoted the expression of YAP and JNK/AP1, leading to the upregulation of the JNK/AP-1 related inflammatory cytokines TNF-α and IL6. Similar results were obtained when osteoblasts were subjected to mechanical stress in vitro. CONCLUSIONS: Our study demonstrated that periodontitis with TO caused severe inflammation-induced bone resorption. Activation of YAP and upregulation of JNK/AP-1 induced by TO potentially aggravated the symptoms of PD.

The comparison of biocompatibility and osteoinductivity between multi-walled and single-walled carbon nanotube/PHBV composites.

The applications of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) in tissue engineering have been widely studied. This study aimed to compare the biocompatibility and osteoinductivity of single-walled carbon nanotubes (SWCNTs)/PHBV composites with multi-walled CNTs (MWCNTs)/PHBV composites. CNTs were dispersed in PHBV by ultrasonication and composites were created using thermal injection moulding. In order to test their biocompatibility and osteoinductivity. Rat osteoblasts (rOBs) were then cultured and seeded on the composites. The composites were implanted in rat femoral bone defects. Our results showed that lower weight percentages of SWCNTs and MWCNTs (2-4%) improved both their mechanical and thermal decomposition properties. However, further reduction of rOBs cell death was observed in MWCNTs/PHBV. SWCNTs were shown to upregulate the expression of Runx-2 and Bmp-2 in early stage significantly, while MWCNTs showed a stronger long-term effect on Opn and Ocn. The in vivo result was that MWCNTs/PHBV composites induced intact rounding new bone, increased integration with new bone, and earlier completed bone remodeling when compared with SWCNTs. Immunohistochemistry also detected higher expression of RUNX-2 around MWCNTs/PHBV composites. In conclusion, there were no differences observed between SWCNTs and MWCNTs in the reinforcement of PHBV, while MWCNTs/PHBV composites showed better biocompatibility and osteoinductivity both in vitro and in vivo.

Inhibition of Ctsk modulates periodontitis with arthritis via downregulation of TLR9 and autophagy.

OBJECTIVES: The mechanisms underlying the effects of Toll-like receptor 9 (TLR9) and autophagy on rheumatoid arthritis (RA)-aggravated periodontitis are unclear. We aimed to explore a novel target, cathepsin K (Ctsk)-mediated TLR9-related autophagy, during the progress of periodontitis with RA. MATERIALS AND METHODS: DBA/J1 mouse model of periodontitis with RA was created by local colonization of Porphyromonas gingivalis (Pg) and injection of collagen. The expression of Ctsk was inhibited by adeno-associated virus (AAV). Micro-CT, immunohistochemistry (IHC), Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to detect the expression of TLR9-related autophagy in periodontitis with RA. Small interfering RNA (siRNA) and CpG oligodeoxynucleotides (CpG ODN) were applied in macrophages. Western blot, immunofluorescence (IF) and qRT-PCR were used to verify the in vivo results. RESULTS: RA can promote periodontitis bone destruction in the lesion area, while inhibiting Ctsk could effectively alleviate this effect. The infiltration of macrophages, TLR9, autophagy proteins (TFEB and LC3) and inflammatory cytokines increased in the periodontitis-with-RA group and was reduced by the inhibition of Ctsk in the periodontal region. Macrophage stimulation confirmed the in vivo results. With the activation of TLR9 by CpG ODN, inhibition of Ctsk could suppress both TLR9 downstream signalling proteins and autophagy-related proteins. CONCLUSIONS: This study advanced a novel role for Ctsk in TLR9 and autophagy to explain the interaction between periodontitis and RA.