Yanmin Zhou

Poly(lactic-co-glycolic) Acid/Nanohydroxyapatite Scaffold Containing Chitosan Microspheres with Adrenomedullin Delivery for Modulation Activity of Osteoblasts and Vascular Endothelial Cells

Adrenomedullin (ADM) is a bioactive regulatory peptide that affects migration and proliferation of diverse cell types, including endothelial cells, smooth muscle cells, and osteoblast-like cells. This study investigated the effects of sustained release of ADM on the modulation activity of osteoblasts and vascular endothelial cells in vitro. Chitosan microspheres (CMs) were developed for ADM delivery. Poly(lactic-co-glycolic) acid and nano-hydroxyapatite were used to prepare scaffolds containing microspheres with ADM. The CMs showed rough surface morphology and high porosity, and they were well-distributed. The scaffolds exhibited relatively uniform pore sizes with interconnected pores. The addition of CMs improved the mechanical properties of the scaffolds without affecting their high porosity. In vitro degradation tests indicated that the addition of CMs increased the water absorption of the scaffolds and inhibited pH decline of phosphate-buffered saline medium. The expression levels of osteogenic-related and angiogenic-related genes were determined in MG63 cells and in human umbilical vein endothelial cells cultured on the scaffolds, respectively. The expression levels of osteogenic-related and angiogenic-related proteins were also detected by western blot analysis. Their expression levels in cells were improved on the ADM delivery scaffolds at a certain time point. The in vitro evaluation suggests that the microsphere-scaffold system is suitable as a model for bone tissue engineering.

Novel bioactive root canal sealer to inhibit endodontic multispecies biofilms with remineralizing calcium phosphate ions

OBJECTIVE The objectives of this study were to: (1) develop a bioactive endodontic sealer via dimethylaminohexadecyl methacrylate (DMAHDM), 2-methacryloyloxyethyl phosphorylcholine (MPC) and nanoparticles of amorphous calcium phosphate (NACP) for the first time; and (2) evaluate inhibition of early-stage and mature multispecies endodontic biofilm, bond strength to root canal dentine, and calcium (Ca) and phosphate (P) ion release. METHODS A series of bioactive endodontic sealers were formulated with DMAHDM, MPC, and NACP. Root dentine bond strength was measured via a push-out test. Three endodontic strains, Enterococcus faecalis, Actinomyces naeslundii, and Fusobacterium nucleatum, were grown on endodontic sealer disks to form multispecies biofilms. Biofilms were grown for 3 days (early) and 14 days (mature). Colony-forming units (CFU), live/dead assay, metabolic activity and polysaccharide were determined. Ca and P ion release from endodontic sealer was measured. RESULTS Incorporating DMAHDM, MPC and NACP did not decrease the push-out bond strength (p>0.1). Adding DMAHDM and MPC reduced endodontic biofilm CFU by 3 log. DMAHDM or MPC each greatly decreased the biofilm CFU (p<0.05). Endodontic sealer with DMAHDM+MPC had much greater killing efficacy than DMAHDM or MPC alone (p<0.05). Endodontic sealer with DMAHDM+MPC had slightly lower, but not significantly lower, Ca and P ion release compared to that without DMAHDM+MPC (p>0.1). CONCLUSIONS A novel bioactive endodontic sealer was developed with potent inhibition of multispecies endodontic biofilms, reducing biofilm CFU by 3 log, while containing NACP for remineralization and possessing good bond strength to root canal dentine walls. CLINICAL SIGNIFICANCE The new bioactive endodontic sealer is promising for endodontic applications to eradicate endodontic biofilms and strengthen root structures. The combination of DMAHDM, MPC and NACP may be applicable to other preventive and restoration resins.

mRNA expression and hypermethylation of tumor suppressor genes apoptosis protease activating factor-1 and death-associated protein kinase in oral squamous cell carcinoma

Apoptosis protease activating factor-1 (Apaf-1) and death-associated protein kinase (DAPK) are p53 pathway-related genes that play significant roles in the activation of caspases, which are involved in mitochondrial-mediated apoptosis. The present study aimed to confirm the role of hyper-methylation of the Apaf-1 and DAPK gene promoter regions in oral squamous cell carcinoma (OSCC) and the effect of the demethylation drug, 5-aza-2′-deoxycytidine (DAC). mRNA from 53 OSCC samples, 23 normal oral mucosa samples and Tca8113 human tongue carcinoma cell lines was detected using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The DNA from each sample was analyzed using methylation-specific PCR (MSP). The Tca8113 cells were demethylated using DAC and the demethylation and re-expression of Apaf-1 and DAPK were analyzed. The Apaf-1 and DAPK mRNA expression index was decreased in 51 (96.23%) and 50 (94.34%) cases, respectively, in the tumor tissues. Hypermethylation of the Apaf-1 and DAPK promoter regions was detected in 46 (86.79%) and 38 (71.69%) cases, respectively. Promoter hypermethylation of the two genes correlated with a decreased mRNA expression in the tumor tissues. Subsequent to being treated with DAC, Apaf-1 and DAPK were demethylated and re-expressed in the Tca8113 cells. Apaf-1 and DAPK promoter hypermethylation may be associated with low gene expression in OSCC. Furthermore, a loss of Apaf-1 and DAPK expression may recover following demethylation. The data provide evidence that methylation exists in OSCC and may play a role in the development of this disease.

Synergistic bactericidal activity of chlorhexidine-loaded, silver-decorated mesoporous silica nanoparticles

Combination of chlorhexidine (CHX) and silver ions could engender synergistic bactericidal effect and improve the bactericidal efficacy. It is highly desired to develop an efficient carrier for the antiseptics codelivery targeting infection foci with acidic microenvironment. In this work, monodisperse mesoporous silica nanoparticle (MSN) nanospheres were successfully developed as an ideal carrier for CHX and nanosilver codelivery through a facile and environmentally friendly method. The CHX-loaded, silver-decorated mesoporous silica nanoparticles (Ag-MSNs@CHX) exhibited a pH-responsive release manner of CHX and silver ions simultaneously, leading to synergistically antibacterial effect against both gram-positive Staphylococcus aureus and gram-negative Escherichia coli. Moreover, the effective antibacterial concentration of Ag-MSNs@CHX showed less cytotoxicity on normal cells. Given their synergistically bactericidal ability and good biocompatibility, these nanoantiseptics might have effective and broad clinical applications for bacterial infections.

Novel multifunctional dental bonding agent for class-V restorations to inhibit periodontal biofilms

We recently developed a dental bonding agent to bond restorations to teeth using nanoparticles of amorphous calcium phosphate (NACP) for remineralization with rechargeable calcium and phosphate ion release. The objectives of this study were to: (1) incorporate an antibacterial monomer dimethylaminohexadecyl methacrylate (DMAHDM) and a protein-repellent agent 2-methacryloyloxyethyl phosphorylcholine (MPC); and (2) investigate protein adsorption and periodontitis-related biofilms for the first time. A primer, used to prime tooth structures for bonding, was made with pyromellitic glycerol dimethacrylate (PMGDM) and 2-hydroxyethyl methacrylate (HEMA). An adhesive was made with PMGDM, ethoxylated bisphenol A dimethacrylate and HEMA. NACP, MPC and DMAHDM were incorporated. Streptococcus gordonii, Actinomyces naeslundii, Porphyromonas gingivalis, Fusobacterium nucleatum were cultured to form single and multi-species biofilms. Colony-forming units (CFU), live/dead, metabolic activity, and polysaccharide were measured. Adding DMAHDM, MPC and NACP into the bonding agent did not compromise the dentin bond strength (p > 0.1). Bonding agents with 5% MPC reduced protein adsorption to 1/15 that of the control (p < 0.05). Bonding agents with 5% DMAHDM + 5% MPC had much greater reduction in biofilms than DMAHDM or MPC alone (p < 0.05). Biofilm CFU was reduced by 3 to 4 log via DMAHDM + MPC. Metabolic activities and polysaccharide of biofilms were also substantially reduced (p < 0.05). In conclusion, a novel bonding agent was developed for dental restorations with inhibition of biofilms, reducing CFU by 3 to 4 log. Besides remineralizartion and acid-neutralization via NACP to inhibit caries as shown previously, the multifunctional adhesive is promising for root restorations with subgingival margins to suppress periodontal pathogens and protect the periodontium.

Poly(L-lactide)-grafted bioglass/poly(lactide-co-glycolide) scaffolds with supercritical CO2 foaming reprocessing for bone tissue engineering

The bioglass particles/poly(lactide-co-glycolide)(BG/PLGA) scaffold has been extensively explored for biomedical applications due to its excellent advantages of mechanical property and controllable degradation rate. In our previous studies, the BG nanoparticle surface-grafted with poly(L-lactide)(PLLA) could substantially improve the phase compatibility between the polymer matrix and the inorganic phase and the biocompatibility of the scaffolds. However, using the traditional preparation methods to prepare the composite scaffold can barely achieve a high porosity and porous connectivity. In this work, the PLLA-grafted bioglass/PLGA(g-BG/PLGA) scaffolds were prepared by supercritical carbon dioxide foaming(Sc-CO2) with before or after particulate leaching(PL) method(Sc-CO2-PL or PL-Sc-CO2 method, PL/Sc-CO2 methods) and their applications in bone replacement and tissue engineering were investigated. The porosities of the g-BG/PLGA scaffolds prepared by the PL/Sc-CO2 methods were higher than 90%, and their mechanical properties had similar values with human cancellous bone. The proliferations of osteoblasts on the scaffolds were dependent on different preparation methods. The PL/Sc-CO2 methods significantly increased the proliferations of the cells. Computed tomography(CT) three-dimensional(3D) reconstruction tomographies of the implantation study for repairing calvarium defects of rabbits demonstrated that the calvarium defects were almost completely filled by the osteotylus in PL/Sc-CO2 method group at 12 week post-surgery, while there was little callus formation in PL method group and untreated control group. These results indicate that the g-BG/PLGA scaffolds prepared by the PL/Sc-CO2 methods exhibit rapid mineralization and osteoconductivity and are the optimal composites for bone repair.

Low-level Ga-Al-As laser irradiation enhances osteoblast proliferation through activation of Hedgehog signaling pathway

Low-level laser irradiation has been reported to promote bone formation, but the molecular mechanism is still unclear. Hedgehog signaling pathway has been reported to play an important role in promoting bone formation. The aim of the present study was to examine whether low-level Ga-Al-As laser (808 nm) irradiation could have an effect on Hedgehog signaling pathway during osteoblast proliferation in vitro. Mouse osteoblastic cell line MC3T3-E1 was cultured in vitro. The cultures after laser irradiation (3.75J/cm2) were treated with recombinant N-terminals Sonic Hedgehog (N-Shh)or Hedgehog inhibitor cyclopamine (cy). The experiment was divided into 4 group, group 1:laser irradiation, group 2: laser irradiation and N-Shh, group 3: laser irradiation and cy, group 4:control with no laser irradiation. On day 1,2 and 3,cell proliferation was determined by cell counting, Cell Counting Kit-8.On 12 h and 24 h, cell cycle was detected by flow cytometry. Proliferation activity of laser irradiation and N-Shh group was remarkably increased compared with those of laser irradiation group. Proliferation activity of laser irradiation and cy group was remarkably decreased compared with those of laser irradiation group, however proliferation activity of laser irradiation and cy group was remarkably increased compared with those of control group. These results suggest that low-level Ga-Al-As laser irradiation activate Hedgehog signaling pathway during osteoblast proliferation in vitro. Hedgehog signaling pathway is one of the signaling pathways by which low-level Ga-Al-As laser irradiation regulates osteoblast proliferation.