Intracellular mRNA phase separation induced by cationic polymers for tumor immunotherapy.

The formation of biomolecular condensates via liquid‒liquid phase separation (LLPS) is an advantageous strategy for cells to organize their subcellular compartments for diverse functions. Recent findings suggest that RNA or RNA-related LLPS techniques have potential for the development of new cellular regulation strategies. However, manipulating RNA LLPS in living cells has great challenges. Herein, we report that cationic polymers (CPs) have strong RNA LLPS-inducing activity. By introducing CPs into living cells or RNA solutions, significant RNA LLPS was verified through confocal imaging, turbidity assays, and fluorescence recovery after photobleaching (FRAP) tests. Among them, turbidity kinetics determinations indicated that the hydrophilic positively charged amino groups on the CPs play essential roles in RNA phase separation. Moreover, the LLPS induced by the cationic polymers dramatically changed the gene expression patterns in the cells. Interestingly, we found that TGFß1 mRNA was highly encapsulated in the RNA droplets, which lowered the immunosuppressive capability of the tumor cells and triggered marked antitumor reactions in a mouse breast cancer model. Thus, we present here the CP-based modulation of RNA LLPS as a novel transcriptional manipulation method with potential for cancer immunotherapy drug development.

Antibacterial and anti-inflammatory activity of valproic acid-pyrazole conjugates as a potential agent against periodontitis.

Periodontitis is a serious global concern. Therefore, in the present study, we intend to synthesize novel valproic-acid pyrazole conjugates as a novel agent against periodontitis. The molecules were developed in a facile synthetic route and obtained in excellent yields. The entire set of molecules were screened for antibacterial activity against a battery of micro-organisms responsible for periodontitis such as P. gingivalis, P. intermedia, F. nucleatum, and E. coli, where they exhibit considerable inhibitory activity. The most potent compound among the tested series, compound 7c showed bactericidal activity in the time-kill curve against E. coli. Compound 7c also showed inhibition of NF-ĸB transcriptional activity in LPS-stimulated RAW264.7 cells with IC50 of 19.23 µM. The effect of compound 7c was also investigated in experimentally induced periodontitis in rats on various indices of oxidative stress (MDA, SOD, and GSH), inflammation (TNF-α, IL-1ß, and IL-6), and apoptosis. It has been found that compound 7c significantly inhibits oxidative stress, inflammation, and apoptosis in a dose-dependent manner. Compound 7c also inhibits the expression of COX-2 and iNOS as shown by western blot analysis.

Thermus caliditerrae sp. nov., a novel thermophilic species isolated from a geothermal area.

Two thermophilic bacterial strains, designated YIM 77925(T) and YIM 77777, were isolated from two hot springs, one in the Hydrothermal Explosion (Shuirebaozhaqu) area and Frog Mouth Spring in Tengchong county, Yunnan province, south-western China. The taxonomic positions of the two isolates were investigated by a polyphasic approach. Cells of the two strains were Gram-stain-negative, aerobic and rod-shaped. They were able to grow at 50-70 °C, pH 6.0-8.0 and with a NaCl tolerance up to 0.5% (w/v). Colonies are circular, convex, non-transparent and produce yellow pigment. Phylogenetic analyses based on 16S rRNA gene sequences comparison clearly demonstrated that strains YIM 77925(T) and YIM 77777 represent members of the genus Thermus, and they also detected low-level similarities of 16S rRNA gene sequences (below 97%) compared with all other species in this genus. Their predominant menaquinone was MK-8. The genomic DNA G+C contents of strains YIM 77925(T) and YIM 77777 were 65.6 mol% and 67.2 mol%, respectively. Based on the results of physiological and biochemical tests and phylogenetic analyses, strains YIM 77925(T) and YIM 77777 could not be classified as representing any species of the genus Thermus with a validly published name. Thus the two strains are considered to represent a novel species of the genus Thermus, for which the name Thermus caliditerrae sp. nov. is proposed. The type strain is YIM 77925(T) ( = DSM 25901(T) = CCTCC 2012061(T)).

Response of microbial communities and biogeochemical cycling functions to sediment physicochemical properties and microplastic pollution under damming and water diversion projects.

Understanding the interactions among flow-sediment, microorganisms, and biogeochemical cycles is crucial for comprehending the ecological response mechanisms of dams and water diversion. This study focused on the spatial patterns of carbon, nitrogen, phosphorus, and sulfur (CNPS) cycle functional genes in the water resource for the middle route of the South-to-North Water Diversion Project in China, specifically the Danjiangkou Reservoir (comprising the Han and Dan reservoirs). The investigation incorporated sediment physicochemical properties and microplastic pollution. Numerous microbial species were identified, revealing that microbial communities demonstrated sensitivity to changes in sedimentary mud content. The communities exhibited greater ß diversity due to finer sediment particles in the Han Reservoir (HR), whereas in the Dan Reservoir (DR), despite having higher sediment nutrient content and MPs pollution, did not display this pattern. Regarding the composition and structure of microbial communities, the study highlighted that sediment N and P content had a more significant influence compared to particle size and MPs. The quantitative microbial element cycling (QMEC) results confirmed the presence of extensive chemolithotrophic microbes and strong nitrogen cycle activity stemming from long-term water storage and diversion operations. The denitrification intensity in the HR surpassed that of the DR. Notably, near the pre-dam area, biological nitrogen fixation, phosphorus removal, and sulfur reduction exhibited noticeable increases. Dam construction refined sediment, fostering the growth of different biogeochemical cycling bacteria and increasing the abundance of CNPS cycling genes. Furthermore, the presence of MPs exhibited a positive correlation with S cycling genes and a negative correlation with C and N cycling genes. These findings suggest that variations in flow-sediment dynamics and MPs pollution have significant impact the biogeochemical cycle of the reservoir.

Disruption of biofilms in periodontal disease through the induction of phase transition by cationic dextrans.

Biofilm of oral pathogenic microorganisms induced by their multiplication and coaggregation would lead to periodontitis. In biofilms, the extracellular polymeric substances (EPS) as a protective shield encapsulates the individual bacteria, protecting them against attack. To alleviate periodontal disease, disrupting the EPS of pathogenic bacteria is crucial and challenging. Based on the sufficient capacity of disorganizing EPS of our designed cationic dextrans, we hypothesized that these polymers could be competent in relieving periodontitis. We validated that cationic dextrans could induce the phase transition of EPS in biofilms, especially the Porphyromonas gingivalis (P. gingivalis), a keystone periodontal pathogen, thus effectively destroying biofilm in vitro. More importantly, satisfactory in vivo treatment was achieved in a rat periodontal disease model. In summary, the study exploited a practical and effective strategy to treat periodontitis with cationic dextrans' powerful biofilm-controlling potential. STATEMENT OF SIGNIFICANCE: Periodontal disease is closely related to dental plaque biofilms on the tooth surface. The biofilm forms gel structures and shields the bacteria underneath, thus protecting oral pathogens from traditional anti-bacterial reagents. Due to limited penetration into gel, the efficacy of these reagents in biofilm elimination is restricted. Our designed cationic dextran could wipe out the coverage of gel-like EPS to disperse encapsulated bacteria. Such superior capacity endowed them with satisfactory effect in disrupting biofilm. Notably, in a rat periodontitis model, cationic dextrans dramatically suppressed alveolar bone loss and alleviated periodontal inflammation by controlling dental plaque. Given the increasing global concerns about periodontal disease, it's worth expanding the application of cationic dextrans both scientifically and clinically.

Symbiotic relationship between Prevotella denticola and Streptococcus mutans enhances virulence of plaque biofilms.

OBJECTIVES: This study aimed to explore that whether interactions between Prevotella denticola and Streptococcus mutans could promote the establishment of hypervirulent biofilms on teeth surface and eventually influence the occurrence and development of caries. DESIGN: Based on single-species biofilms of either P. denticola or S. mutans, and dual-species biofilms of both bacteria, we compared the virulence properties associated with cariogenicity in vitro, including carbohydrate metabolism and acid productivity, synthesis of extracellular polysaccharides, biomass and architecture of biofilms, level of enamel demineralization and expression of virulence genes associated with carbohydrate metabolism and adhesion in S. mutans. RESULTS: The data demonstrated that, compared to single-species of above two taxa, dual-species produced lactate by metabolizing carbohydrates at a higher level during the observation period. Moreover, dual-species biofilms accrued more biomass and exhibited more dense microcolonies and abundant extracellular matrix. And it's noticeable that the level of enamel demineralization in dual-species biofilms was more augmented than that of single-species. In addition, the presence of P. denticola induced the expression of virulence genes gtfs and gbpB in S. mutans. CONCLUSIONS: Symbiotic relationship between P. denticola and S. mutans enhances caries-associated virulence of plaque biofilms, which might provide new strategies for effective prevention and treatment of caries.

Construction of a caries diagnosis model based on microbiome novelty score.

OBJECTIVES: This study aimed to analyze the bacteria in dental caries and establish an optimized dental-ca-ries diagnosis model based on 16S ribosomal RNA (rRNA) data of oral flora. METHODS: We searched the public databa-ses of microbiomes including NCBI, MG-RAST, EMBL-EBI, and QIITA and collected data involved in the relevant research on human oral microbiomes worldwide. The samples in the caries dataset (1 703) were compared with healthy ones (20 540) by using the microbial search engine (MSE) to obtain the microbiome novelty score (MNS) and construct a caries diagnosis model based on this index. Nonparametric multivariate ANOVA was used to analyze and compare the impact of different host factors on the oral flora MNS, and the model was optimized by controlling related factors. Finally, the effect of the model was evaluated by receiver operating characteristic (ROC) curve analysis. RESULTS: 1) The oral microbiota distribution obviously differed among people with various oral-health statuses, and the species richness and species diversity index decreased. 2) ROC curve was used to evaluate the caries data set, and the area under ROC curve was AUC=0.67. 3) Among the five hosts' factors including caries status, country, age, decayed missing filled tooth (DMFT) indices, and sampling site displayed the strongest effect on MNS of samples (P=0.001). 4) The AUC of the model was 0.87, 0.74, 0.74, and 0.75 in high caries, medium caries, low caries samples in Chinese children, and mixed dental plaque samples after controlling host factors, respectively. CONCLUSIONS: The model based on the analysis of 16S rRNA data of oral flora had good diagnostic efficiency.

Variable planning margin approach to account for locoregional variations in setup uncertainties.

PURPOSE: To develop a method for creating variable planning margins around a clinical treatment volume (CTV) and to evaluate its application in head and neck cancer radiotherapy in accounting for locoregional variations of nonrigid setup uncertainties. METHODS: Ten computed tomography (CT) images (with a resolution of 0.68 × 0.68 × 2.5 mm(3)) of a head and neck cancer patient were acquired from the first two weeks of treatment for this study. Five rigid structures (the C2, C5, and caudal C7 vertebrae, mandible, and jugular notch) were used as the landmarks for creating variable local margins. At different CTV locations, local margins were calculated as the weighted average of margins determined at different landmark points from previous studies. The weight was determined by a Gaussian falloff function of the distance between the current location and each landmark point. The CTV delineated on the planning CT image, spanning from the upper portion of the mouth to the lower part of the neck, was expanded to form the planning treatment volume (PTV) with either variable or the conventional constant margins. To evaluate the target coverage, the original planning CTV was deformably mapped to each daily treatment CT using a deformable image registration method. We examined the overlap of the deformed CTV and the rigidly aligned PTV for each margin design strategy and compared the efficacy of the variable margin with the constant margin approach. RESULTS: For the variable margin approach with a baseline C2 margin of 2.5 mm in the left-right, anterior-posterior, and superior-inferior directions, an average of 99.2% of the CTV was within the PTV, and for the approach with a constant 2.5 mm margin, an average of 97.9% of the CTV was within the PTV. With a baseline margin of 2.0 mm, the variable margin approach had an average coverage of 97.8%, similar to that of the constant 2.5 mm margin approach. However, its average nonoverlapped PTV proportion was 32.4%, smaller than that of the constant 2.5 mm margin approach (33.7%). Paired t-tests of computations from the ten treatment fractions showed no significant difference in CTV coverage for the variable margin approach with a baseline of 2.0 mm and the constant 2.5 margin approach (p = 0.054), but the nonoverlapped PTV proportion was significantly smaller for the variable margin approach with a baseline of 2.0 mm than for the constant 2.5 mm margin approach (p < 0.0001). The CTV coverage with the variable margin approach was also significantly higher than with the constant margin approach in the lower neck area, where a larger setup error normally occurs. CONCLUSIONS: We implemented a variable margin approach to account for locoregional variations of setup uncertainties for head and neck cancer radiotherapy, and demonstrated the effectiveness of this approach when compared with the conventional global constant margin expansion approach, where the treatment target spreads out to a broad region. As variable margin data become available and more clinical studies are performed, this approach could be applicable to other treatment sites as well.

Reprograming the immune niche for skin tissue regeneration - From cellular mechanisms to biomaterials applications.

Despite the rapid development of therapeutic approaches for skin repair, chronic wounds such as diabetic foot ulcers remain an unaddressed problem that affects millions of people worldwide. Increasing evidence has revealed the crucial and diverse roles of the immune cells in the development and repair of the skin tissue, prompting new research to focus on further understanding and modulating the local immune niche for comprehensive, 'perfect' regeneration. In this review, we first introduce how different immunocytes and certain stromal cells involved in innate and adaptive immunity coordinate to maintain the immune niche and tissue homeostasis, with emphasis on their specific roles in normal and pathological wound healing. We then discuss novel engineering approaches - particularly biomaterials systems and cellular therapies - to target different players of the immune niche, with three major aims to i) overcome 'under-healing', ii) avoid 'over-healing', and iii) promote functional restoration, including appendage development. Finally, we highlight how these strategies strive to manage chronic wounds and achieve full structural and functional skin recovery by creating desirable 'soil' through modulating the immune microenvironment.

Destructing biofilms by cationic dextran through phase transition.

Eliminating biofilms from infected tissue presents one of the most challenging issues in clinical treatment of chronic wounds. In biofilms, the extracellular polymeric substances (EPS) form gel structures by electrostatic forces between macromolecules. We hypothesized that cationic polymers could induce the gel-to-sol phase transition of the network, leading to biofilms disruptions. We first validated this assumption by using polyethyleneimine (PEI) as a model molecule, and further synthesized two cationic dextrans with high biodegradability for in vitro and in vivo evaluation. All the cationic polymers could destruct Pseudomonas aeruginosa (P. aeruginosa) biofilms. Treating biofilm with cationic dextrans significantly enhanced the bacterial antibiotic sensitivity. When tested in a biofilm-presenting mouse wound healing model, the cationic dextrans efficiently controlled infection, and accelerated the healing process. Our findings suggest that devising cationic polymers to trigger phase transition of biofilm is an effective, straightforward, and perhaps generic strategy for anti-bacterial therapies.

Localized delivery of antisense oligonucleotides by cationic hydrogel suppresses TNF-α expression and endotoxin-induced osteolysis.

PURPOSE: To investigate the possibility of using localized nucleic drug delivery methods for the treatment of osteolysis-related bone disease. METHODS: A bio-degradable cationic hydrogel composed of gelatin and chitosan was used to deliver an antisense oligonucleotide (ASO) targeting murine TNF-α for the treatment of endotoxin-induced osteolysis. RESULTS: ASO combined with this hydrogel was released when it was digested by adhering cells. The released ASO was efficiently delivered into contacted cells and tissues in vitro and in vivo. When tested in animal models of edotoxin-induced bone resorption, ASO delivered by such means effectively suppressed the expression of TNF-α and subsequently the osteoclastogenesis in vivo. Osteolysis in the edotoxin-induced bone resorption animal models was blocked by the treatment. CONCLUSION: This is a successful attempt to apply localized gene delivery method to treat inflammatory diseases in vivo.

Danzhi Jiangtang capsule alleviate hyperglycemia and periodontitis via Wnt/ß-catenin signaling in diabetic rat.

OBJECTIVE: To elucidate whether Danzhi Jiangtang capsule (, DJC) has treatment effects on diabetic periodontitis and the potential mechanism. METHODS: One week after the induction of diabetes by streptozotocin (STZ), 60 male Wistar rats were ligated by orthodontic ligation thread in cervical portion of bilateral maxillary first molar to induce diabetic periodontitis. Periodontitis was exanimated by tooth tissue morphology after 4 weeks. And then all rats were divided into 5 groups: diabetic periodontitis group (DP, n = 20), periodontal basic treatment group (DP + BT, n = 20), periodontal basic treatment +DJC treatment group (DP + BT + DJC, n = 20) and additional Wnt/ß-catenin inhibitor group (DP + BT + DJC + WIKI4/21H7, n = 20). Eight weeks after different interventions, fasting plasma glucose (FPG), C-peptide and glycosylated hemoglobin (HbAlc) of rats were measured and then all rats were sacrificed. The paraffin sections of periodontal tissue were executed hematoxylin-eosin (HE) staining and immunohistochemical examination. The mRNA and protein levels of inflammatory cytokines were evaluated by quantitative polymerase chain reaction (Q-PCR) and Western blot- ting (WB). The protein levels of Wnt/ß-catenin signaling molecules were measured by WB. RESULTS: The blood glucose and C-peptide concentrations in DP, DP + BT and DP + BT + DJC groups were gradually reduced, with gradually decreased distance of CEJ-A and the percentage of periodontal ligament (PDL), as well as gradually increased HbAlc. The number of monocytes and leukocytes in the junctional epithelium and periodontal connective tissue was markedly decreased in DP + BT+ DJC group (P < 0.05), which was slightly reduced in DP+BT group comparing to DP group. The protein levels of Wnt1 and ß-catenin were obviously up-regulated with DJC treatment, while the SOST and DDK1 were markedly down-regulated with DJC treatment. The expression levels of BGP were lowest in DP group and highest in DP + BT + DJC group, while the tumor necrosis factor-α (TNF-α), in- terferon-γ (IFN-γ) and metalloproteinase-3 (MMP-3) were highest in DP group and lowest in DP + BT + DJC group. All these changes could be reversed by Wnt/ß-catenin inhibitor. CONCLUSION: DJC can improve the hyperglycemia and both distal alveolar bone loss and alveolar bone loss in furcation area of diabetic periodontitis rats by reducing the inflammation of gingival tissue and regulating the expressions of BGP, TNF-α, IFN-γ and MMP-3 potentially through Wnt/ß-catenin signaling.

Biomaterial-based physical regulation of macrophage behaviour.

Macrophages play a critical role in regulating immune reactions induced by implanted biomaterials. They are highly plastic and in response to diverse stimuli in the microenvironment can exhibit a spectrum of phenotypes and functions. In addition to biochemical signals, the physical properties of biomaterials are becoming increasingly appreciated for their significant impact on macrophage behaviour, and the underlying mechanisms deserve more in-depth investigations. This review first summarises the effects of key physical cues - including stiffness, topography, physical confinement and applied force - on macrophage behaviour. Then, it reviews the current knowledge of cellular sensing and transduction of physical cues into intracellular signals. Finally, it discusses the major challenges in understanding mechanical regulation that could provide insights for biomaterial design.

A "Bridge-Building" Glycan Scaffold Mimicking Microbial Invasion for In Situ Endothelialization.

The globally high prevalence of peripheral artery diseases poses a pressing need for biomaterials grafts to rebuild vasculature. When implanted, they should promote endothelial cells (ECs) adhesion both profoundly and selectively-but the latter expectation remains unfulfilled. Here, this work is inspired by fungi that invade blood vessels via the "bridge" of galectins that, secreted by ECs, can simultaneously bind carbohydrates on fungal surface and integrin receptors on ECs. A glucomannan decanoate (GMDE) substrate mimicking fungal carbohydrates that highly and preferentially supports ECs adhesion while rejecting several other cell types is designed. Electrospun GMDE scaffolds efficiently sequester endogenous galectin-1-which bridges ECs to the scaffolds as it functions in fungal invasions-and promote blood perfusion in a murine limb ischemic model. Meanwhile, the application of GMDE requires no exogenous pro-angiogenic agents and causes no organ toxicity or adverse inflammation in mice, highlighting its high safety of potential translation. This glycan material, uniquely mimicking a microbial action and harnessing a secreted protein as a "bridge," represents an effective, safe, and different strategy for ischemic vascular therapy.

A novel 3D coordination polymer constructed by dual-ligand for highly sensitive detection of purine metabolite uric acid.

Uric acid (UA), as the final product of purine metabolism, exists in urine and serum, which plays an important role in human metabolism, immunity and other functions. The sensitive, efficient, and rapid detection of UA has far-reaching significance in clinical diagnosis and disease prevention. Herein, a novel coordination polymer constructed by dual-ligand was successfully prepared, which exhibited excellent thermal and water stability. The polymer was interlaced by coordination bonds and hydrogen bonds to form an infinitely extended three-dimensional framework, which showed a rare and novel topological structure. The complex selectively recognized UA through significant fluorescence quenching response in the presence of various interferences. The excellent detection sensitivity (the limited detection of 1.2 µM), outstanding anti-interference ability and remarkable recyclability marked the complex to be a promising sensor material towards UA. In addition, the detection mechanism of UA by the complex was investigated in detail by combining density functional theory (DFT) and a variety of other analytical methods.

Protein corona-guided tumor targeting therapy via the surface modulation of low molecular weight PEG.

The discovery of protein corona (PC) formed on the surface of nanomaterials has promoted research on PC regulation to guide the biological behavior of nanomaterials in vivo. Different from changing the size, shape, and surface charge of nanoparticles, we propose to control the nature of PC by adjusting the molecular weight of low molecular weight polyethylene glycol (LMW PEG, not more than 1000 Da) on the surface of the particles. After excluding the influence of physicochemical factors of PEGylated gold nanoparticles (GNPs), different proteins on the surface of PEGylated GNPs were separated and identified after incubation with human plasma. It is noted that GNP-550 bearing PEG chains of 550 Da absorbed more transferrin responsible for tumor targeting than the other two particles, i.e., GNP-350 and GNP-1000. To validate our speculation, doxorubicin (Dox) was inserted between GNPs and PEGs to explore the cellular and animal studies of Dox-conjugated GNPs. Interestingly, Dox-containing Conj-550 also showed the highest intracellular uptake, cytotoxicity, and apoptosis against HepG2 cells, as well as the best tumor targeting effect and antitumor efficacy in Heps-bearing mice. This protein corona-guided tumor targeting therapy by transferrin provides a new perspective on the function modulation of nanomedicine via LMW PEGs.

Comparative study of covered stent with coil embolization in the treatment of cranial internal carotid artery aneurysm: a nonrandomized prospective trial.

To evaluate whether Willis covered stent implantation yielded angiographic and clinical results were better than those with coil embolization. Eighty-nine patients with cranial internal carotid artery (CICA) aneurysms were treated nonrandomly with covered stents (n = 43, group A) or coil embolization (n = 46, group B). Data on the technical success, procedure time, initial and final angiographic results, and final clinical outcomes were collected and analyzed at >6 months post-procedure. Covered stent placement and coil embolization were successful in all patients, except for one patient in group A. The initial angiographic results showed complete occlusion in 34 group-A patients (80.9%; 95% CI: 69%, 93%) and 24 group-B patients (52.2%; 95% CI: 37%, 67%) (P = 0.004). The final angiographic results indicated complete occlusion in 39 group A patients (39/41, 95.1%; 95% CI: 88%, 102%) and 22 group B patients (48.9%; 95% CI: 34%, 64%) (P < 0.001). The average procedure time was shorter in group A than that in group B (P < 0.001). CICA aneurysm treatment with covered stents yielded better intermediate-term angiographic outcome than those with the recommended approach of coil embolization. (ClinicalTrials.gov number, NCT01029938).

Study on the osteogenesis of rat mesenchymal stem cells and the long-term antibacterial activity of Staphylococcus epidermidis on the surface of silver-rich TiN/Ag modified titanium alloy.

The main causes of failure of orthopedic implants are infection and poor bone ingrowth. Surface modification of the implants to allow for long-term antibacterial and osteogenic functions is an effective solution to prevent failure of the implants. We developed silver-rich TiN/Ag nano-multilayers on the surface of titanium alloy with different doses of Ag+ . The antibacterial stability and osteogenesis of the silver-rich surface were determined by evaluating the adhesion and proliferation of Staphylococcus epidermidis, and the adhesion, proliferation, alkaline phosphatase activity, extracellular matrix mineralization, and the expression level of genes involved in osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs). The results demonstrated that the antibacterial rates (Ra) of 5 × 1016 -Ag, 1 × 1017 -Ag, 5 × 1017 -Ag, and 1 × 1018 -Ag were respectively 46.21%, 85.66%, 94.99%, 98.48%, and 99.99%. After subcutaneous implantation in rats or immersion in phosphate buffered saline for up to 12 weeks, the silver-rich surface of the titanium alloy showed long-term stable inhibition of Staphylococcus epidermidis. Furthermore, in vitro and in vivo studies indicated that the Ag-implanted titanium did not show apparent cytotoxicity and that lower Ag+ implanted groups (5 × 1016 -Ag, 1 × 1017 -Ag) had better viability and biological safety when compared with higher Ag+ implanted groups. In addition, when compared with the Ti6Al4V-group, all Ag-implanted groups exhibited enhanced osteogenic indicators in rat BMSCs. Regarding osteogenic indicators, the surfaces of the 5 × 1017 -Ag group had better osteogenic effects than those of other groups. Therefore, the proper dose of Ag+ implanted TiN/Ag nano-multilayers may be one of the options for the hard tissue replacement materials with antibacterial activity and osteogenic functions.

An "all-in-one" scaffold targeting macrophages to direct endogenous bone repair in situ.

Scaffolds for tissue repair are designed in an increasingly complicated manner to meet multi-facet biological needs during the healing process. However, overly sophisticated design, especially the use of multiple components and delivery of exogenous cells, hampers the bench-to-bedside translation. Here, a multi-functional - yet mono-compositional - bioactive scaffold is devised to mediate the full-range, endogenous bone repair. Based on immunoactivity screening, a chemically-modified glucomannan polysaccharide is selected and processed into an anisotropic porous scaffold, which accurately stimulates macrophages to produce pro-regenerative cytokines. These cytokines effectively enhance the recruitment ("R") and induced osteogenesis ("IO") of the bone progenitor cells in situ. Meanwhile, the anisotropic porosity and carbohydrate signal of the scaffold facilitate differential adhesion ("A") and distribution ("D") of the macrophages and bone progenitor cells - enabling the former's accumulation at the surface while encouraging the latter's infiltration into the scaffold. Implanted in a rat calvarial defect model, this "RADIO" system effectively promotes healing over 12 weeks, with the obvious formation of hard callus through the scaffold. In summary, RADIO integrates multiple functions into one single scalable system ("all-in-one") to govern the dynamic bone-repair process, by harnessing the power of host macrophages. RADIO represents an open platform to solving the long-lasting complexity-versus-simplicity dilemma in biomaterials design. STATEMENT OF SIGNIFICANCE: Biomaterials as versatile tools for tissue repair are becoming increasingly complicated, yet overly sophisticated design - especially the use of multiple components, exogenous cells, and overdosed growth factors - hampers their clinical application. The pre-requisite for designing a successful integrative scaffold is to identify an inherent biological target responding to biomaterial signals, thereby efficiently and safely promoting tissue repair via the endogenous healing capability instead of extra multifarious biochemical components. For bone regeneration, the pivotal regulator is macrophages. Through activating host macrophages, our single-component scaffold system coordinates the entire bone regenerative cascade in situ and induces successful bone regeneration in a calvarial defect model. This scaffold represents a scalable and multi-functional approach to effectively simplify the sophisticated design in regenerative medicine.

Initial Evaluation of a Novel Cone-Beam CT-Based Semi-Automated Online Adaptive Radiotherapy System for Head and Neck Cancer Treatment - A Timing and Automation Quality Study.

Introduction A novel on-line adaptive radiotherapy (ART) system based on O-ring linear accelerator (LINAC) and cone-beam CT (CBCT) was evaluated for treatment and management of head & neck (H&N) cancer in an emulated environment accessed via remote desktop connection. In this on-line ART system, organs-at-risk (OARs) and target contours and radiotherapy (RT) plans are semi-automatically generated based on the patient CBCT, expediting a typically hours-long RT planning session to under half an hour. In this paper, we describe our initial experiences with the system and explore optimization strategies to expedite the process further. Methods We retroactively studied five patients with head and neck cancers, treated 16-35 fractions to 50-70 Gys. For each patient, on-line ART was simulated with one planning CT and three daily CBCT images taken beginning, middle, and end of treatment (tx). Key OAR (mandible, parotids, and spinal cord) and target (planning target volume (PTV) = clinical target volume (CTV) + 3 mm margin) contours were auto-generated and adjusted as needed by therapist/dosimetrist and attending physician, respectively. Duration of OAR contouring, target contouring, and plan review was recorded. Key OAR auto-contours were qualitatively rated from 1 (unacceptable) - 5 (perfect OAR delineation), and then quantitatively compared to human-adjusted "ground truth" contours via dice similarity coefficient (DSC) and 95-percentile Hausdorff distance (HD95%). Once contours were approved, adapted RT plans were auto-generated for physician review. Simulated doses to OARs and targets from the adapted plan were compared to that from the original (un-adapted) plan. Results Median on-line ART planning duration in the remote emulated environment was 19 min 34 sec (range: 13 min 10 sec - 31 min 20 sec). Automated key OAR quality was satisfactory overall (98% scored ≥3; 82% ≥4), though mandible was rated lower than others (p < 0.05). Most key OARs and all targets were within 2 mm margin of human-adjusted contours, but a few parotid and spinal cord contours deviated up to 5 mm. Anatomical changes over tx course further increased auto-contour error (p < 0.05, ΔHD95% = 0.77 mm comparing start and end of tx). Further optimizing auto-contoured OAR and target quality could reduce the on-line treatment planning duration by ~5 min and ~4.5 min, respectively. Dosimetrically, adapted plan spared OARs at a rate much greater than random chance compared to the original plan (χ2 = 22.3, p << 0.001), while maintaining similar therapeutic dose to treatment target CTV (χ2 = 1.14, p > 0.05). In addition, a general decrease in accumulated OAR dose was observed with adaptation. Unsupervised adapted plans where contours were auto-generated without human review still spared OAR at a greater rate than the original plans, suggesting benefits of adaptation can be maintained even with some leniency in contour accuracy. Conclusion Feasibility of a novel, semi-automated on-line ART system for various head and neck (H&N) cancer sites was demonstrated in terms of treatment duration, dosimetric benefits, and automated contour accuracy in a remote emulator environment. Adaptive planning duration was clinically viable at 19 min and 34 sec, but further improvements in automated contour accuracy and performance improvements of plan auto-generation may reduce adaptive planning duration by up to 10 minutes.