Docosahexaenoic Acid-Loaded Nanostructured Lipid Carriers for the Treatment of Peri-Implantitis in Rats.

Being the most common cause of implant failure, peri-implantitis is defined as a pathological condition associated with the occurrence of peri-implant plaque, characterized by peri-implant mucosal inflammation and progressive loss of the supporting bone tissue attributed to the persistence of pro-inflammatory cytokines. Docosahexaenoic acid (DHA), which is a type of omega-3 polyunsaturated fatty acid, is generally used for the treatment of many inflammatory diseases. However, a suitable form for dosing and its therapeutic effect on peri-implantitis remain unclear. In this study, a novel nanostructured lipid carrier (NLC) loaded with squalene and DHA was fabricated (DHA-loaded NLC). The encapsulation efficiency and drug loading efficiency values of the DHA-loaded NLC were 78.13% ± 1.85% and 28.09% ± 0.48%, respectively. The release of DHA was gradual and steady until 144 h. In addition, the free-radical-scavenging rate of DHA-loaded NLC (0.57 ± 0.03) was much higher than that of sole DHA (0.17 ± 0.003). By inhibiting nuclear factor-κB p65 nuclear translocation, DHA-loaded NLC prevented the activation of nuclear factor-κB downstream inflammatory pathways and exerted anti-inflammatory effects on macrophages. Moreover, DHA-loaded NLC showed better effects on preventing alveolar bone resorption of rat peri-implantitis model than sole DHA. Hence, DHA-loaded NLC enhanced the anti-inflammatory bioavailability of DHA, offering a novel approach for the treatment of peri-implantitis.

Multi-functional nanofilms capable of angiogenesis, near-infrared-triggered anti-bacterial activity and inflammatory regulation for infected wound healing.

Chronic infected wound healing is a critical challenge in clinical practice owing to the involvement of multiple physiological processes, including bacteria-related, inflammatory regulation and angiogenesis. Therefore, a multi-functional strategy with synergistic anti-bacterial, anti-inflammatory and pro-angiogenic effects should be developed. Owing to their biomimetic structural features and controlled delivery of active agents, electrospun nanofilms are promising biomaterials for the treatment of skin defects. In this study, we fabricated multi-functional nanofilms with pro-angiogenic, anti-bacterial and anti-inflammatory capacities. First, strontium (Sr) ions were incorporated into poly(L-lactic-co-caprolactone) (PLCL) nanofilms. Subsequently, polydopamine (PDA) and zinc oxide (ZnO) were decorated onto the surface of Sr-loaded PLCL nanofilms to prepare ZnO/PDA@PLCL@Sr nanofilms. In vitro results showed that ZnO/PDA@PLCL@Sr nanofilms were biocompatible, exhibited angiogenic activity and significantly inhibited the growth of Staphylococcus aureus and Escherichia coli upon near-infrared -light irradiation. Furthermore, ZnO/PDA@PLCL@Sr nanofilms were found to drive the transformation of macrophages into the M2 phenotype. In vivo results further validated that ZnO/PDA@PLCL@Sr nanofilms exhibited pro-angiogenic and anti-bacterial activities and regulated inflammation to accelerate wound -healing in a rat model of bacteria-infected skin defects. In conclusion, we successfully developed a multi-functional biomaterial with pro-angiogenic, anti-bacterial and anti-inflammatory capacities to treat chronic infected wounds.

Porphyromonas gingivalis lipopolysaccharide affects oral epithelial connections via pyroptosis.

BACKGROUND/PURPOSE: Pyroptosis is a form of programmed cell death dependent on the activation of caspase-1. Porphyromonas gingivalis (P. gingivalis) is a major pathogenic bacterium in periodontitis and its lipopolysaccharide (LPS) can trigger inflammation. However, whether P. gingivalis-LPS affects epithelial connections or triggers pyroptosis in the gingival epithelium is unknown. MATERIALS AND METHODS: Gingival samples from human donors were collected and the expression levels of E-cadherin, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1/4/5, interleukin (IL)-18, and IL-1ß were examined. P. gingivalis-LPS was injected into rat gingival sulcus to establish gingivitis models, and the expression levels of E-cadherin, NLRP3, caspase-1/11, IL-18, and IL-1ß were compared via immunohistochemistry. The mRNA levels of E-cadherin, caspase-1, IL-18, and IL-1ß were evaluated in oral mucosa epithelial cells (OMECs) and rat gingival tissues. RESULTS: In the present study, NLRP3 (p < 0.01), caspase-1 (p < 0.01), caspase-4 (p = 0.044), and IL-18 (p = 0.036) expression was greater in the human inflammatory gingival samples, whereas E-cadherin (p = 0.045) had the opposite presentation. Gingivitis models were successfully established in rats with the injection of P. gingivalis-LPS. NLRP3 (p = 0.015), caspase-1 (p < 0.01), caspase-11 (p < 0.01), and IL-18 (p = 0.041) were upregulated, whereas E-cadherin (p = 0.038) expression was decreased. Furthermore, E-cadherin mRNA was decreased while caspase-1, IL-18, and IL-1ß mRNA levels were increased. The addition of a caspase-1 inhibitor reversed the expression changes. CONCLUSION: P. gingivalis-LPS may effectively destroy the epithelial connection by triggering pyroptosis.

Keratinized mucosa augmentation guided by double xenogeneic collagen matrix membranes around implants in the posterior mandible: A case report.

RATIONALE: Traditional free gingival graft (FGG) technique is usually used for patients with insufficient peri-implant keratinized mucosa. However, this technique often requires a second surgical area which increases the pain as well as the risk of infection in patients. Xenogeneic collagen matrix (XCM) membrane technique can obtain good results for keratinized mucosa increment. PATIENT CONCERNS: The patient was a 66-year-old healthy female with loss of left mandibular first molar and second molar (FDI #36, #37) for 5 years. Two implants were placed submucosally for 3 months with no interference, while a stage II surgery was needed. DIAGNOSIS: Probing depth measurements suggested that the mesial, medial, and distal widths of buccal keratinized mucosa within the edentulous area were 0.5, 0.5, and 1 mm, respectively, which were insufficient to maintain the health of peri-implant tissues. INTERVENTIONS: Keratinized mucosa augmentation guided by XCM membranes was performed to increase the inadequate buccal keratinized mucosa. OUTCOMES: After 2 months of healing, the widths of mesial, medial, and distal buccal keratinized mucosa were 4, 3, and 3 mm, respectively, and the thickness of the augmented mucosa was 4 mm. Then a stage II surgery was followed. The patient was satisfied with the outcomes of keratinized mucosa augmentation. LESSONS: Keratinized mucosa augmentation guided by double XCM membrane technique can be applied to cases with keratinized mucosa width within 2 mm around implants.

CD44-Targeting Oxygen Self-Sufficient Nanoparticles for Enhanced Photodynamic Therapy Against Malignant Melanoma.

OBJECTIVE: Nanotechnology-based photodynamic therapy (PDT) is a relatively new anti-tumor strategy. However, its efficacy is limited by the hypoxic state in the tumor microenvironment. In the present study, a poly(lactic-co-glycolic acid) (PLGA) nanoparticle that encapsulated both IR820 and catalase (CAT) was developed to enhance anti-tumor therapy. MATERIALS AND METHODS: HA-PLGA-CAT-IR820 nanoparticles (HCINPs) were fabricated via a double emulsion solvent evaporation method. Dynamic light scattering (DLS), transmission electron microscopy (TEM), laser scanning confocal microscopy, and an ultraviolet spectrophotometer were used to identify and characterize the nanoparticles. The stability of the nanoparticle was investigated by DLS via monitoring the sizes and polydispersity indexes (PDIs) in water, PBS, DMEM, and DMEM+10%FBS. Oxygen generation measurement was carried out via visualizing the oxygen bubbles with ultrasound imaging system and an optical microscope. Inverted fluorescence microscopy and flow cytometry were used to measure the uptake and targeting effect of the fluorescent-labeled nanoparticles. The live-dead method and tumor-bearing mouse models were applied to study the HCINP-induced enhanced PDT effect. RESULTS: The results showed that the HCINPs could selectively target melanoma cells with high expression of CD44, and generated oxygen by catalyzing H2O2, which increased the amount of singlet oxygen, ultimately inhibiting tumor growth significantly. CONCLUSION: The present study presents a novel nanoplatform for melanoma treatment.

Effects of D-arginine on Porphyromonas gingivalis biofilm.

Porphyromonas gingivalis (P. gingivalis) is one of the major pathogenic bacteria of periodontitis or peri-implantitis. P. gingivalis tends to attach to the implant's neck with the formation of biofilm, leading to peri-implantitis. d-arginine has been shown to have a potential antimicrobial role. In this study, P. gingivalis was cultured in Brain Heart Infusion broth together with d-arginine. After 3 days (inhibition) or 6 days (dissociation), these were characterized using crystal violet (CV) staining for the biofilm, extracellular polysaccharide (EPS) production from the biofilm, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for biofilm activation. Furthermore, the P. gingivalis biofilm was observed by scanning electron microscopy (SEM). d-arginine effectively reduced biomass accumulation and promoted dissociation at concentrations of ≥50 mM and 100 mM, respectively. Through CV staining, d-arginine concentrations of EPS production from the biofilm for inhibition and dissociation effects was ≥50 mM and 100 mM, respectively. In addition, d-arginine affected biofilm activation for the corresponding concentrations: ≥60 mM for inhibition and ≥90 mM for dispersal. Under SEM observation, d-arginine changed the P. gingivalis biofilm structure in relatively high concentrations for inhibition or dissociation, respectively. The authors concluded that d-arginine could inhibit the formation of P. gingivalis biofilm and promote the dissociation of P. gingivalis biofilm.

Three-Dimensional Porous Scaffolds with Biomimetic Microarchitecture and Bioactivity for Cartilage Tissue Engineering.

Ideal tissue-engineering cartilage scaffolds should possess the same nanofibrous structure as the microstructure of native cartilage as well as the same biological function provided by the microenvironment for neocartilage regeneration. In the present study, three-dimensional composite biomimetic scaffolds with different concentration ratios of electrospun gelatin-polycaprolactone (gelatin-PCL) nanofibers and decellularized cartilage extracellular matrix (DCECM) were fabricated. The nanofibers with the biomimetic microarchitecture of native cartilage served as a skeleton with excellent mechanical properties, and the DCECM served as a biological functionalization platform for the induction of cell response and the promotion of cartilage regeneration. Experimental results showed that the composite nanofiber/DCECM (NF/DCECM) scaffolds had stronger mechanical properties and structural stability in wet state compared with those of DCECM scaffolds. In vitro experiments demonstrated that all scaffolds had good biocompatibility, but the chondrocyte proliferation rate of the composite NF/DCECM scaffolds was higher than that of the NF scaffolds. In vitro and in vivo cartilage regeneration results indicated that the DCECM component of the composite scaffolds facilitated early maturation of the cartilage lacuna and the secretion of collagen and glycosaminoglycan. The macroscopic and histological results at 12 weeks postsurgery exhibited that the composite NF/DCECM scaffolds yielded better cartilage repair outcomes than those of the nontreated group and NF scaffolds group. Overall, the present study demonstrated that the structurally and functionally biomimetic NF/DCECM scaffold is a promising tissue engineering scaffold for cartilage regeneration and cartilage defect repair.

Core­shell type thermo­nanoparticles loaded with temozolomide combined with photothermal therapy in melanoma cells.

A novel core­shell type thermo­nanoparticle (CSTNP) co­loaded with temozolomide (TMZ) and the fluorescein new indocyanine green dye IR820 (termed IT­CSTNPs) was designed and combined with a near­infrared (NIR) laser to realize its photothermal conversion. The IT­CSTNPs were prepared using a two­step synthesis method and comprised a thermosensitive shell and a biodegradable core. IR820 and TMZ were entrapped in the shell and the core, respectively. Dynamic light scattering results demonstrated that the average hydrodynamic size of the IT­CSTNPs was 196.4±3.1 nm with a ζ potential of ­24.9±1.3 mV. The encapsulation efficiencies of TMZ and IR820 were 6.1 and 16.6%, respectively. Temperature increase curves under NIR laser irradiation indicated that the IT­CSTNPs exhibited the desired photothermal conversion efficiency. The in vitro drug release curves revealed a suitable release capability of IT­CSTNP under physiological conditions, whereas NIR laser irradiation accelerated the drug release. Inverted fluorescence microscopy and flow cytometry results revealed that the uptake of IT­CSTNPs by A375 melanoma cells occurred in a concentration­dependent manner. Confocal laser scanning microscopy results indicated that IT­CSTNPs entered tumour cells via endocytosis and were located in intercellular lysosomes. In summary, the present study explored the photothermal conversion capability, cellular uptake, and intracellular localization of IT­CSTNPs.

Experimental study on denitrification using coated electrode of immobilized denitrifying bacteria.

OBJECTIVE: To develop a coated electrode of immobilized denitrificants and to evaluate the performance of a bioelectrochemical reactor to enhance and control denitrification. METHODS: Denitrifying bacteria were developed by batch incubation and immobilized with polyvinyl alcohol (PVA) on the surface of activated carbon fiber (ACF) to make a coated electrode. Then the coated electrode (cathode) and graphite electrode (anode) were transferred to the reactor to reduce nitrate. RESULTS: After acclimated to the mixtrophic and autotrophic denitrification stages, the denitrifying bacteria could use hydrogen as an electron donor to reduce nitrate. When the initial nitrate concentration was 30.2 mg NO3- -N / L, the denitrification efficiency was 57.3% at an applied electric current of 15 mA and a hydraulic retention time (HRT) of 12 hours. Correspondingly, the current density was 0.083 mA/cm2. The nitrate removal rate of the reactor was 34.4 g NO3- -N/m3 x d, and the surface area loading was 1.34 g NO3- -N / m2 x d. CONCLUSION: The coated electrode may keep high quantity of biomass, thus achieving a high denitrification rate. Denitrification efficiencies are related to HRT, current density, oxidation reduction potential (ORP), dissolved oxygen (DO), pH value, and temperature.