Therapeutic Potential of Vasoactive Intestinal Peptide and its Derivative Stearyl-Norleucine-VIP in Inflammation-Induced Osteolysis.

The common use of dental and orthopedic implants calls for special attention to the immune response leading to peri-prosthetic bone loss and implant failure. In addition to the well-established microbial etiology for oral implant failure, wear debris and in particular titanium (Ti) particles (TiP) in the implant vicinity are an important trigger of inflammation and activation of bone resorption around oral and orthopedic implants, presenting an unmet medical need. Here, we employed bacterial-derived lipopolysaccharides (LPS) to model infection and TiP to model aseptic inflammation and osteolysis. We assessed inflammation in vitro by measuring IL1ß, IL6 and TNFα mRNA expression in primary macrophages, osteoclastogenesis in RANKL-induced bone marrow derived pre-osteoclasts and osteolysis in vivo in a mouse calvarial model. We also assessed the trans-epithelial penetrability and safety of the tested compound in rats. Our results show that a lipophilic super-active derivative of vasoactive intestinal peptide (VIP), namely stearyl-norleucine-VIP (SNV) presented superior anti-inflammatory and anti-osteoclastogenic effects compared to VIP in vitro. In the bacterial infection model (LPS), SNV significantly reduced IL1ß expression, while VIP increased IL6 expression. In the aseptic models of osteolysis, SNV showed greater suppression of in vitro osteoclastogenesis than VIP, and significantly inhibited inflammation-induced osteolysis in vivo. We also observed that expression levels of the VIP receptor VPAC-2, but not that of VPAC-1, dramatically decreased during osteoclast differentiation. Importantly, SNV previously shown to have an increased stability compared to VIP, showed here significant trans-epithelial penetration and a clean toxicological profile, presenting a novel drug candidate that could be applied topically to counter both aseptic and infection-related bone destruction.

Author Correction: Bone Anabolic Response in the Calvaria Following Mild Traumatic Brain Injury is Mediated by the Cannabinoid-1 Receptor.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Bone Anabolic Response in the Calvaria Following Mild Traumatic Brain Injury is Mediated by the Cannabinoid-1 Receptor.

Brain trauma was clinically associated with increased osteogenesis in the appendicular skeleton. We showed previously in C57BL/6J mice that mild traumatic brain injury (mTBI) transiently induced bone formation in the femur via the cannabinoid-1 (CB1) receptor. Here, we subjected ICR mice to mTBI and examined the bone response in the skull using microCT. We also measured mast cell degranulation (MCD)72 h post-injury. Finally, we measured brain and calvarial endocannabinoids levels post-mTBI. mTBI led to decreased bone porosity on the contralateral (untouched) side. This effect was apparent both in young and mature mice. Administration of rimonabant (CB1 inverse agonist) completely abrogated the effect of mTBI on calvarial porosity and significantly reduced MCD, compared with vehicle-treated controls. We also found that mTBI resulted in elevated levels of anandamide, but not 2-arachidonoylglycerol, in the contralateral calvarial bone, whereas brain levels remained unchanged. In C57BL/6J CB1 knockout mice, mTBI did not reduce porosity but in general the porosity was significantly lower than in WT controls. Our findings suggest that mTBI induces a strain-specific CB1-dependent bone anabolic response in the skull, probably mediated by anandamide, but seemingly unrelated to inflammation. The endocannabinoid system is therefore a plausible target in management of bone response following head trauma.

Preliminary Study to Evaluate Marginal Bone Loss in Cases of 2- and 3-Implant-Supported Fixed Partial Prostheses of the Posterior Mandible.

PURPOSE: The purpose of this study was to evaluate, by radiographic examinations, the marginal bone resorption around implants in cases of 2 and 3 implant-supported fixed partial prostheses (FPPs) at the posterior mandible. METHODS: A retrospective study of 41 patients (23 males, 18 females) of an average age of 67 years (range, 53-85), with 2 and 3 implants-supported FPPs in the posterior mandible that were treated during 2006 to 2015. The mean follow-up time was 6.32 years (range, 2-10). Twenty-four patients had FPPs on 2 implants (a total of 48 implants). Seventeen patients had FPPs on 3 implants (a total of 51 implants). Clinical and radiographic follow-up examinations were performed. All radiographs were analyzed for changes in marginal bone height surrounding the implants. RESULTS: The mean marginal bone loss around the most mesial implant was slightly higher in the 2-implant group (0.833 mm) compared with the 3-implant group (0.431 mm). The correlation between the mean marginal bone loss around the most mesial implant and the number of implants was of borderline value (P = 0.055). CONCLUSIONS: Considering the limitations of this preliminary study, the authors found that the mesial implant in the 2-implant group is more susceptible to marginal bone loss.

Effect of Two Traditional Polyacrylic Acid Conditioners and 2% Chlorhexidine Digluconate on Cavosurface Microleakage of Glass Ionomer Restorations.

OBJECTIVES: A lack of appropriate adhesiveness is one of the biggest problems in restorative dentistry today and the main cause of microleakage. This is especially true in pediatric dentistry where moisture control is more difficult to achieve. Glass ionomer restorative materials increase adhesion and decrease microleakage given their chemical adhesion to the remaining tooth substance. Pretreatment improves the adhesion quality. The aim of this study was to assess the microleakage of Glass ionomer restorative materials following application of 20% polyacrylic acid, 10% polyacrylic acid or 2% chlorhexidine digluconate in Class V cavities. STUDY DESIGN: Two Class V preparations were prepared on the buccal and lingual surfaces of 24 extracted human molars. The gingival wall was set below or above the CEJ. The teeth were divided into 2 groups. Group 1 was treated with 20% polyacrylic acid or 10% polyacrylic acid. Group 2 was treated with 10% polyacrylic acid or 2% chlorhexidine digluconate. Microleakage was evaluated using a light-reflecting stereomicroscope and stain penetration test. RESULTS: Two percent chlorhexidine digluconate was as efficient as the other conditioners. No statistically significant differences were found among the three types of conditioners. Dye penetration was significantly greater into dentin than into enamel among all three conditioners in both groups (P<0.001). CONCLUSION: Two percent chlorhexidine digluconate, with its known added advantages, can be used as a pretreatment conditioner in GI restorations.

Mechanism and Prevention of Titanium Particle-Induced Inflammation and Osteolysis.

The worldwide number of dental implants and orthopedic prostheses is steadily increasing. Orthopedic implant loosening, in the absence of infection, is mostly attributable to the generation of wear debris. Dental peri-implantitis is characterized by a multifactorial etiology and is the main cause of implant failure. It consists of a peri-implant inflammatory lesion that often results in loss of supporting bone. Disease management includes cleaning the surrounding flora by hand instruments, ultrasonic tips, lasers, or chemical agents. We recently published a paper indicating that US scaling of titanium (Ti) implants releases particles that provoke an inflammatory response and osteolysis. Here we show that a strong inflammatory response occurs; however, very few of the titanium particles are phagocytosed by the macrophages. We then measured a dramatic Ti particle-induced stimulation of IL1ß, IL6, and TNFα secretion by these macrophages using multiplex immunoassay. The particle-induced expression profile, examined by FACS, also indicated an M1 macrophage polarization. To assess how the secreted cytokines contributed to the paracrine exacerbation of the inflammatory response and to osteoclastogenesis, we treated macrophage/preosteoclast cultures with neutralizing antibodies against IL1ß, IL6, or TNFα. We found that anti-TNFα antibodies attenuated the overall expression of both the inflammatory cytokines and osteoclastogenesis. On the other hand, anti-IL1ß antibodies affected osteoclastogenesis but not the paracrine expression of inflammatory cytokines, whereas anti-IL6 antibodies did the opposite. We then tested these neutralizing antibodies in vivo using our mouse calvarial model of Ti particle-induced osteolysis and microCT analysis. Here, all neutralizing antibodies, administered by intraperitoneal injection, completely abrogated the particle-induced osteolysis. This suggests that blockage of paracrine inflammatory stimulation and osteoclastogenesis are similarly effective in preventing bone resorption induced by Ti particles. Blocking both the inflammation and osteoclastogenesis by anti-TNFα antibodies, incorporated locally into a slow-release membrane, also significantly prevented osteolysis. The osteolytic inflammatory response, fueled by ultrasonic scaling of Ti implants, results from an inflammatory positive feedback loop and osteoclastogenic stimulation. Our findings suggest that blocking IL1ß, IL6, and/or TNFα systemically or locally around titanium implants is a promising therapeutic approach for the clinical management of peri-implant bone loss.

Scaling of titanium implants entrains inflammation-induced osteolysis.

With millions of new dental and orthopedic implants inserted annually, periprosthetic osteolysis becomes a major concern. In dentistry, peri-implantitis management includes cleaning using ultrasonic scaling. We examined whether ultrasonic scaling releases titanium particles and induces inflammation and osteolysis. Titanium discs with machined, sandblasted/acid-etched and sandblasted surfaces were subjected to ultrasonic scaling and we physically and chemically characterized the released particles. These particles induced a severe inflammatory response in macrophages and stimulated osteoclastogenesis. The number of released particles and their chemical composition and nanotopography had a significant effect on the inflammatory response. Sandblasted surfaces released the highest number of particles with the greatest nanoroughness properties. Particles from sandblasted/acid-etched discs induced a milder inflammatory response than those from sandblasted discs but a stronger inflammatory response than those from machined discs. Titanium particles were then embedded in fibrin membranes placed on mouse calvariae for 5 weeks. Using micro-CT, we observed that particles from sandblasted discs induced more osteolysis than those from sandblasted/acid-etched discs. In summary, ultrasonic scaling of titanium implants releases particles in a surface type-dependent manner and may aggravate peri-implantitis. Future studies should assess whether surface roughening affects the extent of released wear particles and aseptic loosening of orthopedic implants.