Bioactive fatty acid analog-derived hybrid nanoparticles confer antibody-independent chemo-immunotherapy against carcinoma.

Typical chemo-immunotherapy against malignant carcinoma, is characterized by the combined application of chemotherapeutic agents and monoclonal antibodies for immune checkpoint blockade (ICB). Temporary ICB with antibodies would not depress tumor intrinsic PD-L1 expression and potential PD-L1 adaptive upregulation during chemotherapy, thus exerting limited immunotherapy efficacy. Herein, we developed novel polymer-lipid hybrid nanoparticles (2-BP/CPT-PLNs) for inducing PD-L1 degradation by inhibiting palmitoylation with bioactive palmitic acid analog 2-bromopalmitate (2-BP) to replace PD-L1 antibody (αPD-L1) for ICB therapy, thus achieving highly efficient antitumor immune via immunogenic cell death (ICD) induced by potentiated chemotherapy. GSH-responsive and biodegradable polymer-prodrug CPT-ss-PAEEP10 assisted as a cationic helper polymer could help to stabilize 2-BP/CPT-PLNs co-assembled with 2-BP, and facilitate the tumor site-specific delivery and intracellular release of water-insoluble camptothecin (CPT) in vivo. 2-BP/CPT-PLNs would reinforce cytotoxic CD8+ T cell-mediated antitumor immune response via promoting intratumoral lymphocytes cells infiltration and activation. 2-BP/CPT-PLNs significantly prevented melanoma progression and prolonged life survival of mice beyond the conventional combination of irinotecan hydrochloride (CPT-11) and αPD-L1. Our work first provided valuable instructions for developing bioactive lipid analogs-derived nanoparticles via lipid metabolism intervention for oncotherapy.

Osteopenic effects of high-fat diet-induced obesity on mechanically induced alveolar bone remodeling.

OBJECTIVES: The aim of the study was to investigate the effect of obesity on the tissue and molecular reactions of alveolar bone in response to orthodontic force and its underlying mechanisms. METHODS: Sixty-four rats were randomly divided into normal diet (ND) and high-fat diet (HFD) groups for eight weeks of dietary treatment. OTM was induced using nickel-titanium springs between the upper left first molar and incisor. After 1, 3, 7, and 14 days of OTM, the maxillary alveolar bone and gingival tissues were harvested and analyzed. RESULTS: Compared with the ND rats, the HFD rats had greater OTM distance, serum levels of tartrate-resistant acid phosphatase (TRAP), and tumor necrosis factor α (TNF-α), as well as significant alveolar bone loss and bone architecture deterioration on both the compression and tension sides (p < .05 for all). This response was linked to the increased osteoclast numbers and functional activity and decreased osteoblast activity in the periodontal ligament, gingival tissue, and alveolar bone. CONCLUSIONS: HFD-induced obesity promoted mechanically induced alveolar bone remodeling and detrimental changes in alveolar bone microstructure by increasing osteoclastogenesis and regulating inflammatory cytokine expression. The increased alveolar bone remodeling in the obese rats lead to an accelerated OTM.

Lipid rafts-mediated endocytosis and physiology-based cell membrane traffic models of doxorubicin liposomes.

The clathrin-mediated endocytosis is likely a major mechanism of liposomes' internalization. A kinetic approach was used to assess the internalization mechanism of doxorubicin (Dox) loaded cationic liposomes and to establish physiology-based cell membrane traffic mathematic models. Lipid rafts-mediated endocytosis, including dynamin-dependent or -independent endocytosis of noncaveolar structure, was a dominant process. The mathematic models divided Dox loaded liposomes binding lipid rafts (B) into saturable binding (SB) and nonsaturable binding (NSB) followed by energy-driven endocytosis. The intracellular trafficking demonstrated early endosome-late endosome-lysosome or early/late endosome-cytoplasm-nucleus pathways. The three properties of liposome structures, i.e., cationic lipid, fusogenic lipid, and pegylation, were investigated to compare their contributions to cell membrane and intracellular traffic. The results revealed great contribution of cationic lipid DOTAP and fusogenic lipid DOPE to cell membrane binding and internalization. The valid Dox in the nuclei of HepG2 and A375 cells treated with cationic liposomes containing 40mol% of DOPE were 1.2-fold and 1.5-fold higher than that in the nuclei of HepG2 and A375 cells treated with liposomes containing 20mol% of DOPE, respectively, suggesting the dependence of cell type. This tendency was proportional to the increase of cell-associated total liposomal Dox. The mathematic models would be useful to predict intracellular trafficking of liposomal Dox.

A Dual-Antioxidative Coating on Transmucosal Component of Implant to Repair Connective Tissue Barrier for Treatment of Peri-Implantitis.

Since the microgap between implant and surrounding connective tissue creates the pass for pathogen invasion, sustained pathological stimuli can accelerate macrophage-mediated inflammation, therefore affecting peri-implant tissue regeneration and aggravate peri-implantitis. As the transmucosal component of implant, the abutment therefore needs to be biofunctionalized to repair the gingival barrier. Here, a mussel-bioinspired implant abutment coating containing tannic acid (TA), cerium and minocycline (TA-Ce-Mino) is reported. TA provides pyrogallol and catechol groups to promote cell adherence. Besides, Ce3+ /Ce4+  conversion exhibits enzyme-mimetic activity to remove reactive oxygen species while generating O2 , therefore promoting anti-inflammatory M2 macrophage polarization to help create a regenerative environment. Minocycline is involved on the TA surface to create local drug storage for responsive antibiosis. Moreover, the underlying therapeutic mechanism is revealed whereby the coating exhibits exogenous antioxidation from the inherent properties of Ce and TA and endogenous antioxidation through mitochondrial homeostasis maintenance and antioxidases promotion. In addition, it stimulates integrin to activate PI3K/Akt and RhoA/ROCK pathways to enhance VEGF-mediated angiogenesis and tissue regeneration. Combining the antibiosis and multidimensional orchestration, TA-Ce-Mino repairs soft tissue barriers and effector cell differentiation, thereby isolating the immune microenvironment from pathogen invasion. Consequently, this study provides critical insight into the design and biological mechanism of abutment surface modification to prevent peri-implantitis.

L-arginine protects cementoblasts against hypoxia-induced apoptosis through Sirt1-enhanced autophagy.

BACKGROUND: L-arginine (L-arg) can reduce apoptosis in a variety of cells. Cementoblast apoptosis is related to root resorption during orthodontic treatment. In the present study, we aimed to study the regulatory effect and potential mechanism of L-arg on cementoblast apoptosis and root resorption. METHODS: The apoptosis-related mRNA and protein expression of murine cementoblast (OCCM-30) was assessed after L-arg treatment. To investigate the role of Sirtuin 1 (Sirt1) and autophagy in L-arg resistance to cementoblast apoptosis and root absorption, resveratrol, and EX527 were used to activate or inhibit Sirt1, and chloroquine (CQ) was used to inhibit autophagy. RESULTS: In vitro, L-arg inhibited hypoxia-induced apoptosis in OCCM-30. Further, L-arg increased Sirt1 expression whereas Sirt1 suppression by EX527 reversed the inhibitory effect of L-arg on cell apoptosis. Sirt1 activator resveratrol increased the ratio of microtubule-associated protein light chain 3 (LC3) II/I and decreased the expression of SQSTM1/p62 (p62), suggesting autophagy activation. Autophagy enhancement could reduce apoptosis. Caspase-3 and Bax expression was decreased, and Bcl-2 expression was increased. When autophagy was inhibited by CQ, the positive effects of Sirt1 were attenuated. In vivo, L-arg application reduced root resorption in rats, as demonstrated by decreased root absorption volume. Similarly, L-arg upregulated Sirt1, which activated autophagy in the root resorption model, and less root resorption was observed in the Sirt1 activation group. CONCLUSION: L-arg reduced cementoblast apoptosis in hypoxia and reduced root resorption induced by loading force in rats, which may be partly mediated by Sirt1-enhanced autophagy.

Synthesis of cellulose II-based spherical nanoparticle microcluster adsorbent for removal of toxic hexavalent chromium.

Since natural cellulose is mostly cellulose I and has a fibrous form, most cellulose-based adsorbents are fibrous/rod-shaped and exhibit the cellulose I crystal structure. This study reports a cellulose II-based spherical nanoparticle microcluster adsorbent (SNMA), synthesized from biomass by a bottom-up approach, for removing toxic hexavalent chromium (Cr(VI)). The basic structure of SNMA was investigated. Notably, the prepared adsorbent was a microcluster composed of spherical nanoparticles, while exhibiting cellulose II crystal structure, resulting in higher thermal stability and significantly enhanced adsorption performance. The adsorption process and mechanism of SNMA on Cr(VI) were studied in detail. The SNMA achieved a high adsorption capacity (225.94 mg/g) and receptor site density. The SNMA is expected to be used as a bio-based spherical nanoparticle microcluster adsorbent platform for the adsorption of different toxic substances by changing the surface functional groups of its components, spherical nanoparticles.

Overloading stress-induced progressive degeneration and self-repair in condylar cartilage.

Overloading stress-induced condylar cartilage degeneration acts as the main pathologic change in temporomandibular joint osteoarthritis (TMJ-OA). However, the progression of degeneration and the ability for self-repair remain poorly understood. Here, we explored the progression of cartilage degeneration by dividing pathological stages using a steady mouth-opening mouse model. Then, we observed changes of cartilage by removing the loading at different stages to test the potential self-repair after degeneration induced. Three-dimensional confocal microscopy combined with histology and micro-CT scanning was applied to examine TMJ at different stages of degeneration before and after self-repair. We found the cartilage underwent progressive and thorough degeneration as the overloading stress developed. During the initial adaptation stage, robust proliferation of posteromedial cartilage began at the area of direct loading. Subsequently, widespread chondrocyte apoptosis was found, followed by new chondrocyte proliferation in aggregates with matrix degradation and subchondral bone catabolism. Finally, with cartilage surface damage, the degeneration reached a point where the lesion could not be reversed by self-repair. While the cartilage nearly returned to normal when the interference was removed within 5 days. These results suggested overloading force induces a pathological process of successive degeneration in TMJ cartilage, which can be reversed by self-repair at early stages.

Prevalence and Clinical Correlation of Decayed, Missing, and Filled Teeth in Elderly Inpatients With Schizophrenia.

Introduction: Schizophrenia is a mental disease with a profound impact on human health. Patients with schizophrenia have poor oral hygiene, increasing their risk of systemic diseases, such as respiratory infections, and declining their quality of life. Therefore, this study aims to assess the oral health status of inpatients with schizophrenia, analyze its related factors, and thus provide scientific evidence for further exploration of corresponding control strategies. Methods: A total of 425 inpatients older than 50 years with a diagnosis of schizophrenia from two psychiatric hospitals (mean age 58.49 ± 5.72 years) were enrolled. The demographic data of the patients were checked on admission. Two independent dentists examined caries, missing teeth, and fillings. Mini-Mental State Examination (MMSE) and Global Deterioration Scale were performed as cognitive tests. Positive and Negative Syndrome Scale and Repeatable Battery for the Assessment of Neuropsychological Status rating scale were used to determine their mental status. Results: The average decayed, missing, and filled teeth index was 12.99 ± 8.86. Linear regression analysis showed that the decayed, missing, and filled teeth index had a significantly positive relationship with age (p < 0.001) and smoking (p < 0.001) and a negative relationship with MMSE (p = 0.029). The missing teeth index had a positive relationship with age (p < 0.001), smoking (p < 0.001), and Global Deterioration Scale (p = 0.014) and a negative relationship with MMSE (p = 0.004). Conclusion: The oral health of elderly patients with schizophrenia is poor, which may be related to the cognitive level of patients and affect their quality of life. The focus should be provided to the oral care of patients with schizophrenia, and investment in their specialized oral treatment should be increased.

Chemerin/ChemR23 regulates cementoblast function and tooth resorption in mice via inflammatory factors.

BACKGROUND: Periodontitis and orthodontic treatment can lead to inflammatory root resorption (IRR) through an unclear mechanism. Chemerin, a novel chemoattractant protein, is closely associated with inflammation, affects osteoblast and osteoclast differentiation, and may play a role in IRR. We aimed to explore possible roles of the chemerin/ChemR23 interaction in cementoblast function and IRR and reveal a new IRR therapeutic target. METHODS: Cementoblast function-related gene and protein expression in the immortalized murine cementoblast cell line OCCM-30 after treatment with chemerin and siChemR23 was examined by qRT-PCR and Western blotting. The roles of the MAPK and PI3K-Akt signaling pathways were studied using specific inhibitors. Cementoblast cytokine production under different treatment conditions was measured by enzyme-linked immunosorbent assay and qRT-PCR. Additionally, we modeled IRR in wild-type and chemerin-overexpressing mice and injected transgenic mice with anti-ChemR23 antibody to block ChemR23. We then calculated the root resorption volume and examined periodontal tissue cathepsin K, Runx2, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) expression. RESULT: Chemerin suppressed cementoblast differentiation and mineralization and exerted a proinflammatory effect on cementoblasts. These effects were partially reversed by siChemR23 and reversed to different extents by p38, Erk1/2 and PI3K-Akt pathway inhibition, suggesting p38, Erk1/2 and PI3K-Akt pathways as signaling pathways downstream of chemerin/ChemR23. In vivo, chemerin overexpression worsened IRR. Moreover, chemerin expression was positively correlated with TNF-α, IL-6, and cathepsin K expression and negatively correlated with Runx2 expression. ChemR23 downregulation reversed these effects. CONCLUSION: Chemerin/ChemR23 induced TNF-α and IL-6 expression dependent on Erk1/2, p38 MAPK, and PI3K-Akt signaling pathway activation, thereby regulating cementoblast function and affecting IRR.

D-α-tocopherol polyethylene glycol 1000 succinate-modified liposomes with an siRNA corona confer enhanced cellular uptake and targeted delivery of doxorubicin via tumor priming.

BACKGROUND: Combination therapy employing siRNAs and antitumor drugs is a promising method for the treatment of solid tumors. However, regarding combined treatments involving siRNAs and chemotherapeutic reagents, most prior research has focused on the enhanced cytotoxicity against tumor cells conferred by downregulation of the targeted protein. PURPOSE: We developed D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-modified cationic liposomes (LPs) to simultaneously deliver doxorubicin (Dox) and the Bcl-2 siRNA (siBcl-2) for synergistic chemotherapy. The co-loading of siBcl-2 onto the Dox-loaded cationic LPs (siBcl-2/Dox-TPGS-LPs) could promote cellular uptake, cytotoxicity against 3D H22 tumor spheroids, circulation in the blood, drug accumulation at tumor sites, and synergistic chemotherapy in vivo. METHODS: The siBcl-2/Dox-TPGS-LPs were constructed by co-loading siBcl-2 onto the Dox-loaded TPGS-modified cationic LPs (Dox-TPGS-LPs), and Dox entrapment into the LPs was achieved using an ammonium sulfate gradient method. The antitumor effects of siBcl-2/Dox-TPGS-LPs were studied in murine hepatic carcinoma H22 cells, 3D H22 tumor spheroids, and H22 tumor-bearing mice. RESULTS: Dynamic light scattering technique and transmission electron microscopy images revealed that siBcl-2 loaded onto the Dox-TPGS-LPs formed a prominent corona at an nitrogen to phosphorus (N/P) ratio of 4:1, resulting in particle size increase from 155 to 210 nm and a weak positive zeta potential (+12.5 mV). The siBcl-2/Dox-TPGS-LPs enhanced the cellular uptake of Dox, promoted toxicity against 3D H22 tumor spheroids via tumor priming, prolonged Dox circulation in the blood, and increased accumulation of Dox at tumor sites, thereby enhancing the cytotoxicity of Dox in vitro and its chemotherapeutic efficacy in vivo. CONCLUSION: The siBcl-2/Dox-TPGS-LPs demonstrated a strong potential for application in synergistic chemotherapy. The co-loading of siRNAs both sensitized cells toward antitumor drugs by downregulating the expression level of a specific protein and influenced the pharmacokinetic behavior of the co-delivery system in vitro and in vivo.

Dental Fluorosis over Time: A comparison of National Health and Nutrition Examination Survey data from 2001-2002 and 2011-2012.

Purpose: Excessive fluoride ingestion has been associated with dental fluorosis. The purpose of this study was to determine if there was a difference in dental fluorosis prevalence comparing National Health and Nutrition Examination Survey (NHANES) trend data for adolescents, aged 16 and 17 years, when compared to data collected in 2001-2002 to data from 2011-2012.Methods: The sample included 875 participants. Data analyses included Chi square tests and logistic regressions. The data were from a nationally representative survey by calibrated dental examiners using the modified Dean's fluorosis classification system. The data analysis of the prevalence of fluorosis severity level was dichotomized to very mild/above vs. normal/questionable.Results: In 2001-2002, the weighted percentage prevalence of the denoted dental fluorosis categories were: 49.8% normal (i.e., unaffected), 20.5% questionable, and 29.7% very mild and above. In 2011-2012, the weighted percentage prevalence categories were: 31.2% normal, 7.5% questionable, and 61.3% very mild and above. When comparing years 2001-2002 with the years 2011-2012, the prevalence of very mild and above fluorosis increased by 31.6% (P <.0001) for the 2011-2012 group. In adjusted logistic regression, participants from the years 2011-2012 were more likely to have very mild and above dental fluorosis than participants in 2001-2002 as compared with normal/questionable fluorosis (Adjusted odds ratio= 3.85; 95% confidence interval= 2.20, 6.72; P <.0001).Conclusion: There was a difference of 31.6% in dental fluorosis prevalence between 2012-2011 when compared to data from 2002-2001 in adolescents aged 16 and 17 years. The continued increase in fluorosis rates in the U.S. indicates that additional measures need to be implemented to reduce its prevalence.

Multifunctional hybrid micelles with tunable active targeting and acid/phosphatase-stimulated drug release for enhanced tumor suppression.

Therapeutic efficacy of conventional single PEGylated polymeric micelles is significantly reduced by limited endocytosis and intracellular drug release. To improve drug delivery efficiency, poly (ethylene glycol)-block-poly (l-lactic acid)/(Arg-Gly-Asp-Phe)-poly (aminoethyl ethylene phosphate)-block-poly (l-lactic acid) (PEG-PLLA/RGDF-PAEEP-PLLA) hybrid micelles with tunable active targeting and acid/phosphatase-stimulated drug release are developed. The optimized hybrid micelles with 6 wt % of RGDF have favorable in vitro and in vivo activities. The hybrid micelles could temporarily shield the targeting efficacy of RGDF at pH 7.4 due to the steric effect exerted by concealment of RGDF peptides in the PEG corona, which strongly decreases the clearance by mononuclear phagocyte system and consequently improves the tumor accumulation. Inside the solid tumor with a lower acidic pH, the hybrid micelles restore the active tumor targeting property with exposed RGDF on the surface of the micelles because of the increased protonation and stretching degree of PAEEP blocks. RGDF-mediated endocytosis improves the tumor cell uptake. The hybrid micelles would also enhance intracellular drug release because of the hydrolysis of the acid/phosphatase-sensitivity of PAEEP blocks in endo/lysosome. Systemic administration of the hybrid micelles significantly inhibits tumor growth by 96% due to the integration of enhanced circulation time, tumor accumulation, cell uptake and intracellular drug release.

The association between diabetes mellitus, sugar-sweetened beverages, and tooth loss in adults: Evidence from 18 states.

BACKGROUND: Sugar-sweetened beverages (SSBs) are dietary sources of sugar that are factors in caries development and tooth loss. Dietary sugar also is linked to diabetes mellitus (DM). There is limited research related to SSBs and tooth loss in people with DM. The authors investigated the association between SSBs and tooth loss as it related to the presence or absence of DM. METHODS: The authors used a cross-sectional design with data reported by adults (18 years and older) who responded to the 2012 Behavior Risk Factor Surveillance System questionnaire, which was used in 18 states (N = 95,897; 14,043 who had DM and 81,854 who did not have DM). The authors conducted χ2 and logistic regression analyses to determine associations related to DM status. RESULTS: Overall, 12.3% of the survey respondents had DM, 15.5% had 6 or more teeth extracted, and 22.6% reported that they consumed 1 or more SSB daily. In the adjusted analyses, among adults who had DM, those who consumed at least 2 SSBs daily were more likely to have had 6 or more teeth extracted than those who reported that they did not consume SSBs (adjusted odds ratio, 2.35; 95% confidence interval, 1.37 to 4.01; P = .0018). Among adults who did not have DM, those who consumed more than 1 but fewer than 2 SSBs per day were more likely to have had at least 6 teeth extracted (adjusted odds ratio, 1.46; 95% confidence interval, 1.21 to 1.77; P < .0001). CONCLUSIONS: The authors found that, among adults with DM, consuming 2 or more SSBs per day was associated with having had 6 or more teeth extracted. PRACTICAL IMPLICATIONS: Dietary sugar is a concern for oral and systemic health; however, a strong, independent relationship between the number of teeth extracted and a single source of dietary sugar is not adequate to explain the complexity of tooth loss. Clinicians should use broadly worded dietary messages when discussing caries assessment with patients.

Self-Assembled PAEEP-PLLA Micelles with Varied Hydrophilic Block Lengths for Tumor Cell Targeting.

The properties of hydrophilic shell in micelles significantly affect the interaction between micelles and cells. Compared with frequently used polyethylene glycol (PEG) as the hydrophilic block, polyphosphoesters (PPEs) are superior in functionality, biocompatibility, and degradability. A series of amphiphilic poly(aminoethyl ethylene phosphate)/poly(l-lactide acid) (PAEEP-PLLA) copolymers were synthesized with hydrophilic PAEEP with different chain lengths. The corresponding self-assembled micelles were used for doxorubicin (Dox) entrapment. The length of hydrophilic PAEEP block on the shell affected the structure of micelles. PAEEPm-PLLA168 (m = 130 or 37) polymers formed vesicles, while PAEEPm-PLLA168 (m = 15 or 9) formed large compound micelles (LCMs), suggesting a difference in tumor cell uptake and intracellular trafficking. PAEEP15-PLLA168 polymer showed superiority on cellular uptake amount, intracellular drug release, and cell apoptosis. Lipid rafts and macropinocytosis are the leading endocytic pathways of PAEEP-PLLA micelles. The shape coupling between micelles and cell membrane facilitated cell surface features such as flattened protrusions (membrane protein) and inward-pointing hollows as well as efficient endocytosis. These results suggested that PAEEP-PLLA self-assembled block copolymer micelles may be an excellent drug delivery system for tumor treatment and that the hydrophilic chain length could regulate drug targeting to tumor cells.

Dual-delivery of vancomycin and icariin from an injectable calcium phosphate cement-release system for controlling infection and improving bone healing.

Infectious bone diseases following severely contaminated open fractures are frequently encountered in clinical practice. It is difficult to successfully repair bone and control infection at the same time. To identify a better treatment method, we prepared a dual-drug release system that was comprised of icariin (IC, a natural osteoinductive molecule), vancomycin (VA) and injectable calcium phosphate cement (CPC). The ultrastructure of the dual-drug release system was evaluated by scanning electron microscopy and the biocompatibility was assessed by cell culture. In addition, the release kinetics of IC and VA were respectively investigated by using high­performance liquid chromatography. Finally, this system was used to repair Staphylococcus aureus-contaminated bone defects in a rabbit model. Twelve weeks after the implantation of IC-VA/CPC, the contaminated bone defects were completely repaired, with significantly improved formation of lamellar bone and recanalization of the marrow cavity compared with the controls (CPC without antibiotics or osteoinductive agent). These results demonstrate that this dual-drug release system, with its concomitant antibiotic and osteoinductive properties, has significant potential for the treatment of contaminated bone injury or infectious bone disease.

[An analysis of the characteristics and etiology of hand, foot and mouth disease in a Guangzhou sentinel hospital from May to December, 2008].

OBJECTIVE: To analyze the characteristics and etiology of hand, foot and mouth disease (HFMD) in a sentinel hospital of Guangzhou. METHODS: The epidemiological data and clinical specimens were collected from May to December, 2008 for virological investigations (viral isolation, RT-PCR and molecular identification) and phylogenetic analysis. RESULTS: A total of 309 clinical cases were reported, and the incidence was the highest in 2-4-year-old children, among whom only 15 developed complications, with human enterovirus 71 (HEV71) as the main pathogen (64.7%). Phylogenetic analysis indicated that ten Guangzhou EV71 isolates belonged to Cluster C4a. CONCLUSION: HFMD is an important infectious disease in children resulting from infections by HEV71 as the main pathogen in 2008, and the Guangzhou C4a strains co-evolved with the isolates from other provinces in China and the neighboring countries.