Deep-learning-based ring artifact correction for tomographic reconstruction.

X-ray tomography has been widely used in various research fields thanks to its capability of observing 3D structures with high resolution non-destructively. However, due to the nonlinearity and inconsistency of detector pixels, ring artifacts usually appear in tomographic reconstruction, which may compromise image quality and cause nonuniform bias. This study proposes a new ring artifact correction method based on the residual neural network (ResNet) for X-ray tomography. The artifact correction network uses complementary information of each wavelet coefficient and a residual mechanism of the residual block to obtain high-precision artifacts through low operation costs. In addition, a regularization term is used to accurately extract stripe artifacts in sinograms, so that the network can better preserve image details while accurately separating artifacts. When applied to simulation and experimental data, the proposed method shows a good suppression of ring artifacts. To solve the problem of insufficient training data, ResNet is trained through the transfer learning strategy, which brings advantages of robustness, versatility and low computing cost.

Genomic and metabonomic methods reveal the probiotic functions of swine-derived Ligilactobacillus salivarius.

BACKGROUND: As substitutes for antibiotics, probiotic bacteria protect against digestive infections caused by pathogenic bacteria. Ligilactobacillus salivarius is a species of native lactobacillus found in both humans and animals. Herein, a swine-derived Ligilactobacillus salivarius was isolated and shown to colonize the ileal mucous membrane, thereby promoting nutritional digestion, absorption, and immunity. To evaluate its probiotic role, the entire genome was sequenced, the genetic information was annotated, and the metabolic information was analyzed. RESULTS: The phylogenetic relationship indicated that the bacteria was closer to L. salivarius MT573555.1 and MT585431.1. Functional genes included transporters, membrane proteins, enzymes, heavy metal resistance proteins, and putative proteins; metabolism-related genes were the most abundant. The six types of metabolic pathways secreted by L. salivarius were mainly composed of secretory transmembrane proteins and peptides. The secretory proteins of L. salivarius were digestive enzymes, functional proteins that regulate apoptosis, antibodies, and hormones. Non-targeted metabolomic analysis of L. salivarius metabolites suggested that ceramide, pyrrolidone- 5- carboxylic acid, N2-acetyl-L-ornithine, 2-ethyl-2-hydroxybutyric acid, N-lactoyl-phenylalanine, and 12 others were involved in antioxidation, repair of the cellular membrane, anticonvulsant, hypnosis, and appetite inhibition. Metabolites of clavaminic acid, antibiotic X14889C, and five other types of bacteriocins were identified, namely phenyllactic acid, janthitrem G, 13-demethyl tacrolimus, medinoside E, and tertonasin. The adherence and antioxidation of L. salivarius were also predicted. No virulence genes were found. CONCLUSION: The main probiotic properties of L. salivarius were identified using genomic, metabonomic, and biochemical assays, which are beneficial for porcine feeding. Our results provided deeper insights into the probiotic effects of L. salivarius.

Automatic marker-based alignment method for a nano-resolution full-field transmission X-ray microscope.

Driven by the development of X-ray optics, the spatial resolution of the full-field transmission X-ray microscope (TXM) has reached tens of nanometers and plays an important role in promoting the development of biomedicine and materials science. However, due to the thermal drift and the radial/axial motion error of the rotation stage, TXM computed tomography (CT) data are often associated with random image jitter errors along the horizontal and vertical directions during CT measurement. A nano-resolution 3D structure information reconstruction is almost impossible without a prior appropriate alignment process. To solve this problem, a fully automatic gold particle marker-based alignment approach without human intervention was proposed in this study. It can automatically detect, isolate, and register gold particles for projection image alignment with high efficiency and accuracy, facilitating a high-quality tomographic reconstruction. Simulated and experimental results confirmed the reliability and robustness of this method.

Feature detection network-based correction method for accurate nano-tomography reconstruction.

Driven by the development of advanced x-ray optics such as Fresnel zone plates, nano-resolution full-field transmission x-ray microscopy (Nano-CT) has become a powerful technique for the non-destructive volumetric inspection of objects and has long been developed at different synchrotron radiation facilities. However, Nano-CT data are often associated with random sample jitter because of the drift or radial/axial error motion of the rotation stage during measurement. Without a proper sample jitter correction process prior to reconstruction, the use of Nano-CT in providing accurate 3D structure information for samples is almost impossible. In this paper, to realize accurate 3D reconstruction for Nano-CT, a correction method based on a feature detection neural network, which can automatically extract target features from a projective image and precisely correct sample jitter errors, is proposed, thereby resulting in high-quality nanoscale 3D reconstruction. Compared with other feature detection methods, even if the target feature is overlapped by other high-density materials or impurities, the proposed Nano-CT correction method still acquires sub-pixel accuracy in geometrical correction and is more suitable for Nano-CT reconstruction because of its universal and faster correction speed. The simulated and experimental datasets demonstrated the reliability and validity of the proposed Nano-CT correction method.

RNA-binding protein RALY reprogrammes mitochondrial metabolism via mediating miRNA processing in colorectal cancer.

OBJECTIVE: Dysregulated cellular metabolism is a distinct hallmark of human colorectal cancer (CRC). However, metabolic programme rewiring during tumour progression has yet to be fully understood. DESIGN: We analysed altered gene signatures during colorectal tumour progression, and used a complex of molecular and metabolic assays to study the regulation of metabolism in CRC cell lines, human patient-derived xenograft mouse models and tumour organoid models. RESULTS: We identified a novel RNA-binding protein, RALY (also known as hnRNPCL2), that is highly associated with colorectal tumour aggressiveness. RALY acts as a key regulatory component in the Drosha complex, and promotes the post-transcriptional processing of a specific subset of miRNAs (miR-483, miR-676 and miR-877). These miRNAs systematically downregulate the expression of the metabolism-associated genes (ATP5I, ATP5G1, ATP5G3 and CYC1) and thereby reprogramme mitochondrial metabolism in the cancer cell. Analysis of The Cancer Genome Atlas (TCGA) reveals that increased levels of RALY are associated with poor prognosis in the patients with CRC expressing low levels of mitochondrion-associated genes. Mechanistically, induced processing of these miRNAs is facilitated by their N6-methyladenosine switch under reactive oxygen species (ROS) stress. Inhibition of the m6A methylation abolishes the RALY recognition of the terminal loop of the pri-miRNAs. Knockdown of RALY inhibits colorectal tumour growth and progression in vivo and in organoid models. CONCLUSIONS: Collectively, our results reveal a critical metabolism-centric role of RALY in tumour progression, which may lead to cancer therapeutics targeting RALY for treating CRC.

Deep-learning-based image registration for nano-resolution tomographic reconstruction.

Nano-resolution full-field transmission X-ray microscopy has been successfully applied to a wide range of research fields thanks to its capability of non-destructively reconstructing the 3D structure with high resolution. Due to constraints in the practical implementations, the nano-tomography data is often associated with a random image jitter, resulting from imperfections in the hardware setup. Without a proper image registration process prior to the reconstruction, the quality of the result will be compromised. Here a deep-learning-based image jitter correction method is presented, which registers the projective images with high efficiency and accuracy, facilitating a high-quality tomographic reconstruction. This development is demonstrated and validated using synthetic and experimental datasets. The method is effective and readily applicable to a broad range of applications. Together with this paper, the source code is published and adoptions and improvements from our colleagues in this field are welcomed.

Curcumin-Loaded Blood-Stable Polymeric Micelles for Enhancing Therapeutic Effect on Erythroleukemia.

Curcumin has high potential in suppressing many types of cancer and overcoming multidrug resistance in a multifaceted manner by targeting diverse molecular targets. However, the rather low systemic bioavailability resulted from its poor solubility in water and fast metabolism/excretion in vivo has hampered its applications in cancer therapy. To increase the aqueous solubility of curcumin while retaining the stability in blood circulation, here we report curcumin-loaded copolymer micelles with excellent in vitro and in vivo stability and antitumor efficacy. The two copolymers used for comparison were methoxy-poly(ethylene glycol)-block-poly(ε-caprolactone) (mPEG-PCL) and N-(tert-butoxycarbonyl)-l-phenylalanine end-capped mPEG-PCL (mPEG-PCL-Phe(Boc)). In vitro cytotoxicity evaluation against human pancreatic SW1990 cell line showed that the delivery of curcumin in mPEG-PCL-Phe(Boc) micelles to cancer cells was efficient and dosage-dependent. The pharmacokinetics in ICR mice indicated that intravenous (i.v.) administration of curcumin/mPEG-PCL-Phe(Boc) micelles could retain curcumin in plasma much better than curcumin/mPEG-PCL micelles. Biodistribution results in Sprague-Dawley rats also showed higher uptake and slower elimination of curcumin into liver, lung, kidney, and brain, and lower uptake into heart and spleen of mPEG-PCL-Phe(Boc) micelles, as compared with mPEG-PCL micelles. Further in vivo efficacy evaluation in multidrug-resistant human erythroleukemia K562/ADR xenograft model revealed that i.v. administration of curcumin-loaded mPEG-PCL-Phe(Boc) micelles significantly delayed tumor growth, which was attributed to the improved stability of curcumin in the bloodstream and increased systemic bioavailability. The mPEG-PCL-Phe(Boc) micellar system is promising in overcoming the key challenge of curcumin's to promote its applications in cancer therapy.

Hair regrowth in alopecia areata patients following Stem Cell Educator therapy.

BACKGROUND: Alopecia areata (AA) is one of the most common autoimmune diseases and targets the hair follicles, with high impact on the quality of life and self-esteem of patients due to hair loss. Clinical management and outcomes are challenged by current limited immunosuppressive and immunomodulating regimens. METHODS: We have developed a Stem Cell Educator therapy in which a patient's blood is circulated through a closed-loop system that separates mononuclear cells from the whole blood, allows the cells to briefly interact with adherent human cord blood-derived multipotent stem cells (CB-SC), and returns the "educated" autologous cells to the patient's circulation. In an open-label, phase 1/phase 2 study, patients (N = 9) with severe AA received one treatment with the Stem Cell Educator therapy. The median age was 20 years (median alopecic duration, 5 years). RESULTS: Clinical data demonstrated that patients with severe AA achieved improved hair regrowth and quality of life after receiving Stem Cell Educator therapy. Flow cytometry revealed the up-regulation of Th2 cytokines and restoration of balancing Th1/Th2/Th3 cytokine production in the peripheral blood of AA subjects. Immunohistochemistry indicated the formation of a "ring of transforming growth factor beta 1 (TGF-ß1)" around the hair follicles, leading to the restoration of immune privilege of hair follicles and the protection of newly generated hair follicles against autoimmune destruction. Mechanistic studies revealed that co-culture with CB-SC may up-regulate the expression of coinhibitory molecules B and T lymphocyte attenuator (BTLA) and programmed death-1 receptor (PD-1) on CD8ß(+)NKG2D(+) effector T cells and suppress their proliferation via herpesvirus entry mediator (HVEM) ligands and programmed death-1 ligand (PD-L1) on CB-SCs. CONCLUSIONS: Current clinical data demonstrated the safety and efficacy of the Stem Cell Educator therapy for the treatment of AA. This innovative approach produced lasting improvement in hair regrowth in subjects with moderate or severe AA. TRIAL REGISTRATION: ClinicalTrials.gov, NCT01673789, 21 August 2012.

Roles of hydroxyapatite allocation and microgroove dimension in promoting preosteoblastic cell functions on photocured polymer nanocomposites through nuclear distribution and alignment.

This study clarifies how hydroxyapatite (HA) allocation and microgroove dimension affect mouse preosteoblastic MC3T3-E1 cell functions on microgrooved substrates of polymer nanocomposites. Using replica molding from micromachined silicon wafer templates, we fabricated photocured poly(ε-caprolactone) triacrylate (PCLTA)/HA nanocomposite substrates with parallel microgrooves (two groove widths of 5 and 15 µm and one groove depth of 5 µm). Four types of microgrooved substrates were made: "homogeneous" ones of PCLTA and PCLTA/HA with uniform distribution and two "heterogeneous" laminated microgrooved substrates with HA only in the PCLTA matrix in the ridges or bottom. These substrates were used to regulate MC3T3-E1 cell attachment, proliferation, alignment, nuclear circularity and distribution, and mineralization. MC3T3-E1 cell attachment and proliferation were much higher on the microgrooved substrates of PCLTA/HA than on those of PCLTA, in particular, on the 5 µm wide microgrooved substrate with PCLTA/HA ridges and PCLTA bottom. The shape and distribution of MC3T3-E1 cytoskeleton and nuclei were altered by the substrate topography and HA allocation. For 5 µm wide heterogeneous microgrooved substrates with HA only in the ridges, MC3T3-E1 cells exhibited better spreading perpendicular to the microgrooves but tended to extend along the microgrooves containing HA in the bottom. The widest cells and the roundest/largest cell nuclei were observed on the heterogeneous substrate with PCLTA/HA ridges, while the narrowest cells with the best elongation were found on the homogeneous PCLTA/HA substrate. The trend in MC3T3-E1 cell mineralization on the substrates was consistent with that in cell/nuclear elongation. Osteocalcin mRNA expression was significantly higher on the PCLTA/HA substrates than on the PCLTA ones and also on the microgrooved substrates of PCLTA/HA than on the flat ones, regardless of the groove width of 5 or 15 µm.

SUMO-1 Promotes Ishikawa Cell Proliferation and Apoptosis in Endometrial Cancer by Increasing Sumoylation of Histone H4.

OBJECTIVES: To investigate the functional role of SUMO-1 on cell proliferation and apoptosis in endometrial cancer cells (Ishikawa line) and to explore the underlying regulatory mechanisms. METHODS: Different concentrations of estradiol (E2) and small interfering RNA (siRNA) targeting the SUMO-1 (siCo) were treated in Ishikawa cells, and then quantitative polymerase chain reaction was used to examine the expression of progesterone receptor (PR) expression in Ishikawa cells. Western blots were applied to validate histone H4 sumoylation. CCK8 assay and flow cytometry were performed to investigate cell proliferation and apoptosis in Ishikawa cells. RESULTS: Estradiol up-regulated the expression of PR messenger RNA, most obviously at 100 nM. SUMO-1 siRNA decreased the PR expression. Estradiol up-regulated H4 sumoylation and caused the increase of Ishikawa cell proliferation, whereas SUMO-1 siRNA down-regulated H4 sumoylation, inhibited the cell proliferation, and induced apoptosis. Furthermore, SUMO-1 siRNA-transfected cells were arrested in the S and G2/M phases and E2 increased the S and G2/M phases of Ishikawa cells. CONCLUSION: SUMO-1 regulates the Ishikawa cell proliferation and apoptosis by the sumoylation of histone H4.

Opposite Mechanical Preference of Bone/Nerve Regeneration in 3D-Printed Bioelastomeric Scaffolds/Conduits Consistently Correlated with YAP-Mediated Stem Cell Osteo/Neuro-Genesis.

To systematically unveil how substrate stiffness, a critical factor in directing cell fate through mechanotransduction, correlates with tissue regeneration, novel biodegradable and photo-curable poly(trimethylene carbonate) fumarates (PTMCFs) for fabricating elastomeric 2D substrates and 3D bone scaffolds/nerve conduits, are presented. These substrates and structures with adjustable stiffness serve as a unique platform to evaluate how this mechanical cue affects the fate of human umbilical cord mesenchymal stem cells (hMSCs) and hard/soft tissue regeneration in rat femur bone defect and sciatic nerve transection models; whilst, decoupling from topographical and chemical cues. In addition to a positive relationship between substrate stiffness (tensile modulus: 90-990 kPa) and hMSC adhesion, spreading, and proliferation mediated through Yes-associated protein (YAP), opposite mechanical preference is revealed in the osteogenesis and neurogenesis of hMSCs as they are significantly enhanced on the stiff and compliant substrates, respectively. In vivo tissue regeneration demonstrates the same trend: bone regeneration prefers the stiffer scaffolds; while, nerve regeneration prefers the more compliant conduits. Whole-transcriptome analysis further shows that upregulation of Rho GTPase activity and the downstream genes in the compliant group promote nerve repair, providing critical insight into the design strategies of biomaterials for stem cell regulation and hard/soft tissue regeneration through mechanotransduction.

Targeting insulin resistance in type 2 diabetes via immune modulation of cord blood-derived multipotent stem cells (CB-SCs) in stem cell educator therapy: phase I/II clinical trial.

BACKGROUND: The prevalence of type 2 diabetes (T2D) is increasing worldwide and creating a significant burden on health systems, highlighting the need for the development of innovative therapeutic approaches to overcome immune dysfunction, which is likely a key factor in the development of insulin resistance in T2D. It suggests that immune modulation may be a useful tool in treating the disease. METHODS: In an open-label, phase 1/phase 2 study, patients (N=36) with long-standing T2D were divided into three groups (Group A, oral medications, n=18; Group B, oral medications+insulin injections, n=11; Group C having impaired ß-cell function with oral medications+insulin injections, n=7). All patients received one treatment with the Stem Cell Educator therapy in which a patient's blood is circulated through a closed-loop system that separates mononuclear cells from the whole blood, briefly co-cultures them with adherent cord blood-derived multipotent stem cells (CB-SCs), and returns the educated autologous cells to the patient's circulation. RESULTS: Clinical findings indicate that T2D patients achieve improved metabolic control and reduced inflammation markers after receiving Stem Cell Educator therapy. Median glycated hemoglobin (HbA1C) in Group A and B was significantly reduced from 8.61%±1.12 at baseline to 7.25%±0.58 at 12 weeks (P=2.62E-06), and 7.33%±1.02 at one year post-treatment (P=0.0002). Homeostasis model assessment (HOMA) of insulin resistance (HOMA-IR) demonstrated that insulin sensitivity was improved post-treatment. Notably, the islet beta-cell function in Group C subjects was markedly recovered, as demonstrated by the restoration of C-peptide levels. Mechanistic studies revealed that Stem Cell Educator therapy reverses immune dysfunctions through immune modulation on monocytes and balancing Th1/Th2/Th3 cytokine production. CONCLUSIONS: Clinical data from the current phase 1/phase 2 study demonstrate that Stem Cell Educator therapy is a safe approach that produces lasting improvement in metabolic control for individuals with moderate or severe T2D who receive a single treatment. In addition, this approach does not appear to have the safety and ethical concerns associated with conventional stem cell-based approaches. TRIAL REGISTRATION: ClinicalTrials.gov number, NCT01415726.

HRD1 functions as a tumor suppressor in ovarian cancer by facilitating ubiquitination-dependent SLC7A11 degradation.

The E3 ubiquitin ligase 3-hydroxy-3-methylglutaryl reductase degradation (HRD1) was found to be a tumor suppressor in diverse types of cancers; we aimed to explore its expression pattern and biological function in ovarian cancer (OC). HRD1 expression in OC tumor tissues was detected using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). The overexpression plasmid of HRD1 was transfected into OC cells. Cell proliferation, colony formation, and apoptosis were analyzed using bromodeoxy uridineassay, colony formation assay, and flow cytometry, respectively. OC mice models were established to explore the effect of HRD1 on OC in vivo. Ferroptosis was evaluated by malondialdehyde, reactive oxygen species, and intracellular ferrous iron. Expressions offerroptosis-related factors were examined using qRT-PCR and western blot. Erastin and Fer-1 were, respectively, employed to promote or inhibit ferroptosis in OC cells. Online bioinformatics tool and co-immunoprecipitation assay were performed to predict and verify the interactive genes of HRD1 in OC cells, respectively. Gain-of-function studies were carried out to determine the roles of HRD1 in cell proliferation, apoptosis, and ferroptosis in vitro. HRD1 was under-expressed in OC tumor tissues. The overexpression of HRD1 inhibited OC cell proliferation and colony formation in vitro and suppressed OC tumor growth in vivo. The overexpression of HRD1 promoted cell apoptosis and ferroptosis in OC cell lines. HRD1 interacted with the solute carrier family 7 member 11 (SLC7A11) in OC cells, and HRD1 regulated the stability and ubiquitination in OC. SLC7A11 overexpression recovered the effect of HRD1 overexpression in OC cell lines. HRD1 inhibited tumor formation and promoted ferroptosis in OC through enhancing SLC7A11 degradation.

Device-Performance-Driven Heterogeneous Multiparty Learning for Arbitrary Images.

Multiparty learning (MPL) is an emerging framework for privacy-preserving collaborative learning. It enables individual devices to build a knowledge-shared model and remaining sensitive data locally. However, with the continuous increase of users, the heterogeneity gap between data and equipment becomes wider, which leads to the problem of model heterogeneous. In this article, we concentrate on two practical issues: data heterogeneous problem and model heterogeneous problem, and propose a novel personal MPL method named device-performance-driven heterogeneous MPL (HMPL). First, facing the data heterogeneous problem, we focus on the problem of various devices holding arbitrary data sizes. We introduce a heterogeneous feature-map integration method to adaptively unify the various feature maps. Meanwhile, to handle the model heterogeneous problem, as it is essential to customize models for adapting to the various computing performances, we propose a layer-wise model generation and aggregation strategy. The method can generate customized models based on the device's performance. In the aggregation process, the shared model parameters are updated through the rules that the network layers with the same semantics are aggregated with each other. Extensive experiments are conducted on four popular datasets, and the result demonstrates that our proposed framework outperforms the state of the art (SOTA).

Poly(ε-caprolactone)-banded spherulites and interaction with MC3T3-E1 cells.

We report that protein adsorption, cell attachment, and cell proliferation were enhanced on spherulites-roughened polymer surfaces. Banded spherulites with concentric alternating succession of ridges and valleys were observed on spin-coated thin films of poly(ε-caprolactone) (PCL) and two series of PCL binary homoblends composed of high- and low-molecular-weight components when they were isothermally crystallized at 25-52 °C. Their thermal properties, crystallization kinetics, and surface morphology were examined. The melting temperature (T(m)), crystallinity (χ(c)), crystallization rate, and spherulitic patterns showed strong dependence on the crystallization temperature (T(c)) and the blend composition. The surface roughness of the spherulites was higher when T(c) was higher; thus, the larger surface area formed in banded spherulites could adsorb more serum proteins from cell culture media. In vitro mouse preosteoblastic MC3T3-E1 cell attachment, proliferation, and nuclear localization were assessed on the hot-compressed flat disks and spherulites-roughened films of the high-molecular-weight PCL and one of its homoblends. The number of attached MC3T3-E1 cells and the proliferation rate were greater on the rougher surfaces than those on the flat ones. It is interesting to note that cell nuclei were preferentially, though not absolutely, located in or close to the valleys of the banded spherulites. The percentage of cell nuclei in the valleys was higher than 78% when the ridge height and adjacent ridge distance were ~350 and ~35 nm, respectively. This preference was weaker when the ridge height was lower or at a higher cell density. These results suggest that isothermal crystallization of semicrystalline polymers can be an effective thermal treatment method to achieve controllable surface roughness and pattern for regulating cell behaviors in tissue-engineering applications.

Photocured biodegradable polymer substrates of varying stiffness and microgroove dimensions for promoting nerve cell guidance and differentiation.

Photocross-linkable and biodegradable polymers have great promise in fabricating nerve conduits for guiding axonal growth in peripheral nerve regeneration. Here, we photocross-linked two poly(ε-caprolactone) triacrylates (PCLTAs) with number-average molecular weights of ~7000 and ~10,000 g mol(-1) into substrates with parallel microgrooves. Cross-linked PCLTA7k was amorphous and soft, while cross-linked PCLTA10k was semicrystalline with a stiffer surface. We employed different dimensions of interests for the parallel microgrooves, that is, groove widths of 5, 15, 45, and 90 µm and groove depths of 0.4, 1, 5, and 12 µm. The behaviors of rat Schwann cell precursor line (SpL201) cells with the glial nature and pheochromocytoma (PC12) cells with the neuronal nature were studied on these microgrooved substrates, showing distinct preference to the substrates with different mechanical properties. We found different threshold sensitivities of the two nerve cell types to topographical features when their cytoskeleton and nuclei were altered by varying the groove depth and width. Almost all of the cells were aligned in the narrowest and deepest microgrooves or around the edge of microgrooves. Oriented SpL201 cell movement had a higher motility as compared to unaligned ones. After forskolin treatment, SpL201 cells demonstrated significantly upregulated S-100 and O4 on stiffer substrates or narrower microgrooves, suggesting more differentiation toward early Schwann cells (SCs). PC12 neurites were oriented with enhanced extension in narrower microgrooves. The present results not only improve our fundamental understanding on nerve cell-substrate interactions, but also offer useful conduit materials and appropriate feature dimensions to foster guidance for axonal growth in peripheral nerve regeneration.

Optimal poly(L-lysine) grafting density in hydrogels for promoting neural progenitor cell functions.

Recently, we have developed a photopolymerizable poly(L-lysine) (PLL) that can be covalently incorporated into poly(ethylene glycol) diacrylate (PEGDA) hydrogels to improve their bioactivity by providing positive charges. To explore the potential of these PLL-grafted PEGDA hydrogels as a cell delivery vehicle and luminal filler in nerve guidance conduits for peripheral and central nerve regeneration, we varied the number of pendent PLL chains in the hydrogels by photo-cross-linking PEGDA with weight compositions of PLL (φ(PLL)) of 0, 1, 2, 3, and 5%. We further investigated the effect of PLL grafting density on E14 mouse neural progenitor cell (NPC) behavior including cell viability, attachment, proliferation, differentiation, and gene expression. The amount of actually grafted PLL and charge densities were characterized, showing a proportional increase with the feed composition φ(PLL). NPC viability in 3D hydrogels was significantly improved in a PLL grafting density-dependent manner at days 7 and 14 postencapsulation. Similarly, NPC attachment and proliferation were promoted on the PLL-grafted hydrogels with increasing φ(PLL) up to 2%. More intriguingly, NPC lineage commitment was dramatically altered by the amount of grafted PLL chains in the hydrogels. NPC differentiation demonstrated a parabolic or nonmonotonic dependence on φ(PLL), resulting in cells mostly differentiated toward mature neurons with extensive neurite formation and astrocytes rather than oligodendrocytes on the PLL-grafted hydrogels with φ(PLL) of 2%, whereas the neutral hydrogels and PLL-grafted hydrogels with higher φ(PLL) of 5% support NPC differentiation less. Gene expression of lineage markers further illustrated this trend, indicating that PLL-grafted hydrogels with an optimal φ(PLL) of 2% could be a promising cell carrier that promoted NPC functions for treatment of nerve injuries.

Promoting nerve cell functions on hydrogels grafted with poly(L-lysine).

We present a novel photopolymerizable poly(L-lysine) (PLL) and use it to modify polyethylene glycol diacrylate (PEGDA) hydrogels for creating a better, permissive nerve cell niche. Compared with their neutral counterparts, these PLL-grafted hydrogels greatly enhance pheochromocytoma (PC12) cell survival in encapsulation, proliferation, and neurite growth and also promote neural progenitor cell proliferation and differentiation capacity, represented by percentages of both differentiated neurons and astrocytes. The role of efficiently controlled substrate stiffness in regulating nerve cell behavior is also investigated and a polymerizable cationic small molecule, [2-(methacryloyloxy)ethyl]-trimethylammonium chloride (MTAC), is used to compare with this newly developed PLL. The results indicate that these PLL-grafted hydrogels are promising biomaterials for nerve repair and regeneration.

Lubricated biodegradable polymer networks for regulating nerve cell behavior and fabricating nerve conduits with a compositional gradient.

We present a method of tuning surface chemistry and nerve cell behavior by photo-cross-linking methoxy poly(ethylene glycol) monoacrylate (mPEGA) with hydrophobic, semicrystalline poly(ε-caprolactone) diacrylate (PCLDA) at various weight compositions of mPEGA (ø(m)) from 2 to 30%. Improved surface wettability is achieved with corresponding decreases in friction, water contact angle, and capability of adsorbing proteins from cell culture media because of repulsive PEG chains tethered in the network. The responses of rat Schwann cell precursor line (SpL201), rat pheochromocytoma (PC12), and E14 mouse neural progenitor cells (NPCs) to the modified surfaces are evaluated. Nonmonotonic or parabolic dependence of cell attachment, spreading, proliferation, and differentiation on ø(m) is identified for these cell types with maximal values at ø(m) of 5-7%. In addition, NPCs demonstrate enhanced neuronal differentiated lineages on the mPEGA/PCLDA network at ø(m) of 5% with intermediate wettability and surface energy. This approach lays the foundation for fabricating heterogeneous nerve conduits with a compositional gradient along the wall thickness, which are able to promote nerve cell functions within the conduit while inhibiting cell attachment on the outer wall to prevent potential fibrous tissue formation following implantation.

Synergistic effect of the anti-PD-1 antibody with blood stable and reduction sensitive curcumin micelles on colon cancer.

Curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione) is a potent anticancer drug with versatile biological activities, while the clinical translation of curcumin is severely limited due to its hydrophobicity, rapid elimination, and metabolism in the blood circulation. Herein, we aim to unravel the potential of curcumin as a synergistic agent with immunotherapy in the treatment of cancers. In an effort to minimize premature release and improve the systemic bioavailability, a superior blood stable and reduction sensitive curcumin micellar formulation, of which the release can be triggered by cancer cells, is rationally designed. We have synthesized a telodendrimer (mPEG-PLA-(LA)4) capable of forming reversible disulfide crosslinked micelles (DCMs). The curcumin loaded DCMs (Cur/DCMs) are spherical with a uniform size of 24.6 nm. The in vitro release profile demonstrates that curcumin releases significantly slower from DCMs than that from non-crosslinked micelles (NCMs), while the release can be accelerated with the increasing concentration of reducing agent glutathione (GSH). Intravenous administration of Cur/DCMs stably retains curcumin in the bloodstream and efficiently improves the systemic bioavailability. Furthermore, Cur/DCMs exhibit synergistic anticancer efficacy when combined with the anti-PD-1 antibody in an MC-38 colon cancer xenograft model. Our results potentiate the integration of blood stable curcumin nanoformulation and immunotherapy for cancer treatment.