Development of an antimicrobial peptide-loaded mineralized collagen bone scaffold for infective bone defect repair.

The repair of infective bone defects is a great challenge in clinical work. It is of vital importance to develop a kind of bone scaffold with good osteogenic properties and long-term antibacterial activity for local anti-infection and bone regeneration. A porous mineralized collagen (MC) scaffold containing poly(d,l-lactide-co-glycolic acid) (PLGA) microspheres loaded with two antibacterial synthetic peptides, Pac-525 or KSL-W was developed and characterized via scanning electron microscopy (SEM), porosity measurement, swelling and mechanical tests. The results showed that the MC scaffold embedded with smooth and compact PLGA microspheres had a positive effect on cell growth and also had antibacterial properties. Through toxicity analysis, cell morphology and proliferation analysis and alkaline phosphatase evaluation, the antibacterial scaffolds showed excellent biocompatibility and osteogenic activity. The antibacterial property evaluated with Staphylococcus aureus and Escherichia coli suggested that the sustained release of Pac-525 or KSL-W from the scaffolds could inhibit the bacterial growth aforementioned in the long term. Our results suggest that the antimicrobial peptides-loaded MC bone scaffold has good antibacterial and osteogenic activities, thus providing a great promise for the treatment of infective bone defects.

Integrative Analysis for Elucidating Transcriptomics Landscapes of Glucocorticoid-Induced Osteoporosis.

Osteoporosis is the most common bone metabolic disease, characterized by bone mass loss and bone microstructure changes due to unbalanced bone conversion, which increases bone fragility and fracture risk. Glucocorticoids are clinically used to treat a variety of diseases, including inflammation, cancer and autoimmune diseases. However, excess glucocorticoids can cause osteoporosis. Herein we performed an integrated analysis of two glucocorticoid-related microarray datasets. The WGCNA analysis identified 3 and 4 glucocorticoid-related gene modules, respectively. Differential expression analysis revealed 1047 and 844 differentially expressed genes in the two datasets. After integrating differentially expressed glucocorticoid-related genes, we found that most of the robust differentially expressed genes were up-regulated. Through protein-protein interaction analysis, we obtained 158 glucocorticoid-related candidate genes. Enrichment analysis showed that these genes are significantly enriched in the osteoporosis related pathways. Our results provided new insights into glucocorticoid-induced osteoporosis and potential candidate markers of osteoporosis.

Comprehensive Analysis of the Genetic and Epigenetic Mechanisms of Osteoporosis and Bone Mineral Density.

Osteoporosis is a skeletal disorder characterized by a systemic impairment of bone mineral density (BMD). Genome-wide association studies (GWAS) have identified hundreds of susceptibility loci for osteoporosis and BMD. However, the vast majority of susceptibility loci are located in non-coding regions of the genome and provide limited information about the genetic mechanisms of osteoporosis. Herein we performed a comprehensive functional analysis to investigate the genetic and epigenetic mechanisms of osteoporosis and BMD. BMD and osteoporosis are found to share many common susceptibility loci, and the corresponding susceptibility genes are significantly enriched in bone-related biological pathways. The regulatory element enrichment analysis indicated that BMD and osteoporosis susceptibility loci are significantly enriched in 5'UTR and DNase I hypersensitive sites (DHSs) of peripheral blood immune cells. By integrating GWAS and expression Quantitative Trait Locus (eQTL) data, we found that 15 protein-coding genes are regulated by the osteoporosis and BMD susceptibility loci. Our analysis provides new clues for a better understanding of the pathogenic mechanisms and offers potential therapeutic targets for osteoporosis.

A Mendelian Randomization Analysis to Expose the Causal Effect of IL-18 on Osteoporosis Based on Genome-Wide Association Study Data.

Accumulating evidence showed that Interleukin (IL) level is associated with Osteoporosis. Whereas, most of these associations are based on observational studies. Thus, their causality was still unclear. Mendelian randomization (MR) is a widely used statistical framework that uses genetic instrumental variables (IVs) to explore the causality of intermediate phenotype with disease. To classify their causality, we conducted a MR analysis to investigate the effect of IL-18 level on the risk of Osteoporosis. First, based on summarized genome-wide association study (GWAS) data, 8 independent IL-18 SNPs reaching genome-wide significance were deemed as IVs. Next, Simple median method was used to calculate the pooled odds ratio (OR) of these 8 SNPs for the assessment of IL-8 on the risk of Osteoporosis. Then, MR-Egger regression was utilized to detect potential bias due to the horizontal pleiotropy of these IVs. As a result of simple median method, we get the SE (-0.001; 95% CI-0.002 to 0; P = 0.042), which means low IL-18 level could increases the risk of the development of Osteoporosis. The low intercept (0; 95% CI -0.001 to 0; P = 0.59) shows there is no bias due to the horizontal pleiotropy of the IVs.

Recurrent sialoliths after excision of the bilateral submandibular glands for sialolithiasis treatment: A case report.

Sialolithiasis is a common disease that is characterized by the obstruction of the salivary gland. Sialolithiasis mainly affects the submandibular glands and the Wharton's duct. However, bilateral sialolithiasis is a rare condition. In addition, recurrence of sialoliths subsequent to surgical excision of the submandibular gland for the treatment of sialolithiasis has been rarely reported. The present study reported a case presenting with recurrent sialoliths with sialadenitis in the residual Wharton's duct following the excision of bilateral submandibular glands. An 81-year-old man presented with a solid and painful mass in the left submandibular area. The patient had a history of bilateral submandibular sialolithiasis, and had undergone excision of bilateral submandibular glands with the right Wharton's duct 4 years earlier. Computed tomography scans demonstrated two calculi in the residual Wharton's duct, which were surgically removed without any complications. The present study discussed the mechanisms underlying sialolith formation subsequent to the excision of submandibular glands.

Effects of fluoride-ion-implanted titanium surface on the cytocompatibility in vitro and osseointegatation in vivo for dental implant applications.

As an attractive technique for the improvement of biomaterials, Plasma immersion ion implantation (PIII) has been applied to modifying the titanium material for dental implant application. The present study investigated the cytocompatibility and early osseointegration of fluoride-ion-implanted titanium (F-Ti) surface and implants, both characterizing in their composition of titanium oxide and titanium fluoride. The cytocompatibility of F-Ti was evaluated in vitro by using scanning electron microscope, Cell Counting Kit-8 assay, alkaline phosphatase activity assay, and quantitative real-time polymerase chain reaction. The results showed that the F-Ti weakened the effects that Porphyromonas gingivalis exerted on the MG-63 cells in terms of morphology, proliferation, differentiation, and genetic expression when MG-63 cells and Porphyromonas gingivalis were co-cultured on the surface of F-Ti. Meanwhile, the osteogenic activity of F-Ti implants was assessed in vivo via evaluating the histological morphology and estimating histomorphometric parameters. The analysis of toluidine blue staining indicated that the new bone was more mature in subjects with F-Ti group, which exhibited the Haversian system, and the mean bone-implant contact value of F-Ti group was slightly higher than that of cp-Ti group (p>0.05). Fluorescence bands were wider and brighter in the F-Ti group, and the intensity of fluorochromes deposited at the sites of mineralized bone formation was significantly higher for F-Ti surfaces than for cp-Ti surfaces, within the 2nd, 3rd and 4th weeks (p<0.05). An indication is that the fluoride modified titanium can promote cytocompatibility and early osseointegration, thus providing a promising alternative for clinical use.

High in vitro antibacterial activity of Pac-525 against Porphyromonas gingivalis biofilms cultured on titanium.

In order to investigate the potential of short antimicrobial peptides (AMPs) as alternative antibacterial agents during the treatment of peri-implantitis, the cytotoxic activity of three short AMPs, that is, Pac-525, KSL-W, and KSL, was determined using the MTT assay. The antimicrobial activity of these AMPs, ranging in concentration from 0.0039 mg/mL to 0.5 mg/mL, against the predominant planktonic pathogens, including Streptococcus sanguis, Fusobacterium nucleatum, and Porphyromonas gingivalis, involved in peri-implantitis was investigated. Furthermore, 2-day-old P. gingivalis biofilms cultured on titanium surfaces were treated with Pac-525 and subsequently observed and analysed using confocal laser scanning microscopy (CLSM). The average cell proliferation curve indicated that there was no cytotoxicity due to the three short AMPs. The minimum inhibitory concentration and minimum bactericidal concentration values of Pac-525 were 0.0625 mg/mL and 0.125 mg/mL, respectively, for P. gingivalis and 0.0078 mg/mL and 0.0156 mg/mL, respectively, for F. nucleatum. Using CLSM, we confirmed that compared to 0.1% chlorhexidine, 0.5 mg/mL of Pac-525 caused a significant decrease in biofilm thickness and a decline in the percentage of live bacteria. These data indicate that Pac-525 has unique properties that might make it suitable for the inhibition the growth of pathogenic bacteria around dental implants.

[Correlation analysis of EphA7 expression with clinico-pathological parameters and prognosis in tongue squamous cell carcinoma].

PURPOSE: To evaluate the relationship between the expression of EphA7 and its clinical correlation and function with tongue squamous cell carcinoma (TSCC). METHODS: The expression of EphA7 was determined in 54 pairs of human TSCC tissues and pair-matched adjacent noncancerous tissues by immunohistochemistry. Furthermore, association between EphA7 expression and patients' clinicopathological characteristics, overall and disease-free survival were evaluated. Invasion and metastasis of SCC9 cell were detected before and after down regulation of EphA7 expression. Differences in measurement data were compared with paired-t test, and survival analysis was made by Kaplan-Meier method using SPSS17.0 software package. RESULTS: EphA7 was positive in all examined specimens. Significant associations were noted between high EphA7 expression and absence of lymph node metastasis, absence of vascular invasion, dense stromal inflammatory reaction and female gender. TSCC patients with higher EphA7 expression presented longer overall and disease-free survival compared with low EphA7 expression. The invasion and metastasis of SCC9 cell increased significantly after down regulation of EphA7. CONCLUSIONS: The study indicated that EphA7 may participate in the malignant transformation of TSCC, reinforcing their utility as targets for potential therapeutic intervention.

Bio-Oss(®) for delayed osseointegration of implants in dogs: a histological study.

We evaluated the effects of Bio-Oss® (a natural bone substitute derived from the mineral portion of bovine bone) on delayed osseointegration of implants. The bilateral third and fourth mandibular premolars of 4 adult, healthy, male and female dogs were extracted. We randomly selected 2 extraction sockets in each dog to be filled with Bio-Oss® (the experimental group); the other 2 extraction sockets, which were not treated, served as controls. Dental implants were inserted into the alveolar bone of the experimental group and the control group 3 months after insertion of the Bio-Oss®. The osteogenic activity in the bone around the implants was assessed by evaluating the histological morphology and estimating histomorphometric variables at 3 and 6 months after delayed implantation. After 3 months, Goldner's trichrome staining analysis showed that the rate of content between the bone and the implant and the mineralised area of bone around the implant were significantly higher in the experimental group (76%(9%) and 69.5% (9.6%), respectively) than those in the control group (56.1% (8.2%) and 52.8% (7.3%), respectively, p=0.003 and 0.000). However, the 2 groups did not differ significantly at 6 months. Fluorescence microscopy showed that the mean rates of mineralisation of the bony tissue around the implant in the experimental group at months 3 and 6 were 6.8 (0.4) µm and 8.4 (0.8) µm, respectively, which were significantly higher than those in the control group (p=0.000 and 0.03). These data indicate that putting Bio-Oss® into the extraction sockets can promote osseointegration after delayed implantation, and may be a promising option for clinical use.