Sequential Release Research Articles

Sequential Release of Ibuprofen and the Gasotransmitter Hydrogen sulfide using Oxanorbornane-Based Synthetic Lipids as Carriers.

After understanding the biological signaling roles of hydrogen sulfide and its involvement in various physiological processes, there has been enormous interest in exploring its therapeutic utility in areas such as cancer, inflammation, cardiovascular diseases, etc. There is also growing interest in using suitable H2S donors in combination with other drugs to improve the treatment outcome through the modulation of multiple pathways. The premature release of H2S from small molecule donors and the difficulty in controlling its spatio-temporal distribution are the major challenges during these efforts. Hence the development of appropriate carriers that can release this gasotransmitter along with the therapeutic entity of interest in a controlled manner has high significance. In this regard, this report presents a novel drug delivery system from oxanorbornane-based synthetic lipids that carries a H2S-releasing 1,2-dithiole-3-thione moiety as part of the head group. Nanoaggregates of the resulting conjugate are not only capable of efficiently entrapping a non-steroidal anti-inflammatory drug such as ibuprofen, but also release this drug and H2S in a controlled and sequential manner.

Medial soft tissue release is also related to the anterior stability of cruciate-retaining total knee arthroplasty: a cadaveric study.

Medial soft tissue release is occasionally performed to achieve mediolateral ligament balance in total knee arthroplasty (TKA), whose sequential effect on mediolateral and anteroposterior stability remains unclear. This study aimed to quantitatively evaluate the difference in mediolateral and anteroposterior stability according to a sequential medial soft tissue release in TKA. Cruciate-retaining TKA was performed in six cadaveric knees. Medial and lateral joint gaps, varus-valgus angle, and tibial anterior and posterior translations relative to the femur with pulling and pushing forces, respectively, were measured. All measurements were performed at full extension and 45° and 90° flexion after release of the deep medial collateral ligament (MCL) (stage 1), the posteromedial capsule (stage 2), and the superficial MCL (stage 3). Mediolateral and anteroposterior stability were compared between stages, and correlations between mediolateral and anteroposterior stability were analyzed. Medial joint gap significantly increased from stages 1 to 3 by 3.2mm, 6.8mm, and 7.2mm at extension, 45° flexion, and 90° flexion, respectively, and from stages 2 to 3 by 3.5mm at extension. Varus-valgus angle was varus at stage 2, which turned to valgus at stage 3 (-2.7° to 0.8°, -2.2° to 4.3°, and -5.5° to 2.5° at extension, 45° flexion, and 90° flexion, respectively). Anterior translation at 90° flexion significantly increased from stages 1 and 2 to stage 3 by 11.5mm and 8.2mm, respectively, which was significantly correlated with medial gap (r = 0.681) and varus-valgus angle (r = 0.495). Medial soft tissue release also increased tibial anterior translation as well as medial joint gap, and medial joint gap and tibial anterior translation were significantly correlated. Surgeons should be careful not to create too large medial joint gap and tibial anterior translation in flexion by excessive medial release up to the superficial MCL for achieving an equal mediolateral joint gap in extension.

Revolutionising High Resolution HLA Genotyping for Transplant Assessment: Validation, Implementation and Challenges of Oxford Nanopore Technologies' Q20+ Sequencing.

The advent of third-generation sequencing (TGS) represents a significant shift in the field of genetic sequencing, enabling single-molecule sequencing to overcome limitations of short-read NGS platforms. Several studies have assessed the utilisation of TGS in HLA genotyping, though many of these studies have described the high error rate as an obstacle to achieving a robust and highly accurate HLA typing assay. In 2021, Oxford Nanopore Technologies (ONT) released the high-accuracy sequencing Kit 14 and the MinION flow cell model R10.4.1, which were reported to achieve sequencing accuracies up to 99%. The aim of this study was to validate this novel high-accuracy sequencing kit for HLA genotyping coupled with a full-gene HLA PCR assay. Comparison with historical data obtained using legacy flow cell models such as R9.4, R10.3 and R10.4 was also done to assess progressive improvement in sequencing performance with each sequential release. The workflow was validated based on data throughput, sequence quality and accuracy, and HLA genotyping resolution. An initial validation was performed using an internal reference panel of 42 samples representing common HLA allele groups, followed by an analysis of data obtained from 111 sequencing batch runs since the implementation, to assess assay performance and define quality control metrics to assess inter-run variability and monitor quality. Furthermore, challenges arising from MinION flow cell stability and use, and assessment of barcode contamination are discussed. The findings of this study highlight advantages of ONT sequencing kit 14/R10.4.1 for HLA genotyping and the implementation considerations for the routine diagnostic HLA laboratory.

Redox-responsive degradation of antimicrobials with programmable drug release for enhanced antibacterial activity

The global crisis of antibiotic resistance has impelled the exigency to develop more effective drug delivery systems for the treatment of bacterial infection. The development of possessing high biocompatibility and targeted delivery of antimicrobials remains a persisting challenge. For programmable release of efficient antimicrobials in infection sites to enhance antibacterial activity, herein, we fabricated diselenide-bridged mesoporous organosilica nanoparticle-supported silver nanoparticles (Ag NPs) with high drug-loading capacity for the co-delivery of tobramycin (TOB) within one drug delivery system (Ag-MON@TOB (Se)). The resultant Ag-MON@TOB (Se) exhibited favorable biocompatibility due to its high stability in the physiological condition. Notably, such Ag-MON@TOB (Se) manifested a programmable structural destabilization to trigger sequential drug release in response to the oxidative stimuli within the bacterial infection microenvironment. In contradistinction to the oxidation-stable disulfide bond moieties within the framework of the nanocarrier (Ag-MON@TOB (S)), the Ag-MON@TOB (Se) with its programmed drug release behavior augmented prominent antibacterial therapy both in vitro and in vivo. This work represents a promising strategy for programmable drug release by harnessing a responsive degradable vehicle to enhance the treatment of bacterial infection.

A multi-step mathematical model-based predictive strategy for software release timing during testing stage

Mathematically precise modeling is important to be established to accurately examine the quantitative relationship between software testing and software reliability. Software testing process is complex since it is concerned with various factors such as test case execution, defect debugging, tester expertise, test case selection, and so forth. For this reason, it is required to be meticulous in formulating the software testing process in a manner which is mathematically concise. The software release life cycle or sequential release timeline, referring to the process related to the development, testing and distribution of a software product comprises several critical stages, and the length of this particular life cycle reveals variations depending on different factors like the type of product, the intended use of it, industry security, general standards and compliance. One consideration software engineers have is related to the release date of the software so that future commitments about the software’s release time can be formulated beforehand. In view of these aspects, a multi-step strategy for predicting software release dates is proposed in the current study along with the following stages: firstly, the proposed technique selects the utmost reliability growth model that very well fits the observed test data halfway through the testing period, and then employs it to forecast the probable date of release. This technique entails approximating the unknown parameters of suitable Software Reliability Growth Models (SRGMs). Finally, the chosen SRGM is used to forecast the release date of the software under test by fitting it to available fault data. The proposed method is straightforward and applied to test on a total of ten actual datasets collected from the literature. The results of the proposed technique reveal that in the majority of the situations, nearly exact approximation of date of release can be made halfway through the testing period. Moreover, the proposed method’s performance is also compared to that of a number of previous strategies present in the literature. The outcomes obtained by our study demonstrate that the proposed strategy may be used to forecast the release date of software in practical situations.

Sequential release of vancomycin and BMP-2 from chitosan/nano-hydroxyapatite thermosensitive hydrogel for the treatment of chronic osteomyelitis

In this study, we developed scaffolds materials with microspheres to form a double sustained release system.Chitosan/nano-hydroxyapatite (CS-HA) was used as a drug carrier to construct a sustained-release system for Bone morphogenetic protein-2(BMP-2) and Vancomycin (VAN). Furthermore, VAN and BMP-2 loaded microspheres (Ms) were prepared by the emulsion ultrasonic method.The resultant composites were characterized by Scanning electron microscope (SEM), compressive strength, porosity, and biodegradation. The characterization results showed uniform porous and rough surface, enhanced thermal stability, and highest compressive strength ((1.912 ± 0.012) Kpa, the surface of the two microspheres was slightly folded and showed a regular spherical shape.The loading rate of BMP-2 was (59.611 × 10-4 ± 0.023 × 10-4)% and the encapsulation rate was (6.022 ± 0.005)%. The release rate of vancomycin and BMP-2 was 57.194% and 12.968% respectively. Osteogenic differentiation of Bone marrow mesenchymal stem cells (BMSCs) was confirmed by alkaline phosphatase quantification. The deposition of late osteogenic markers (calcium phosphates) detected by Alizarin red, which indicated extracellular matrix mineralization. The results showed that BMP-2/VAN in CS-HA hydrogel successfully achieved the sequential release of the double drugs, which could benefit bone regeneration.

Euler Force-Driven Siphon Valve Control for Precise Sequential Release in Centrifugal Microfluidic Chips

Controlling the fluids in centrifugal microfluidic chips for precise sequential release is critical for multi-step reactions and immunoassays. Currently, the traditional methods of liquid sequential release mainly rely on various types of microvalves, which face the problems of complex operation and high costs. Here, this work presents a method for driving liquid release using the Euler force. Under continuous acceleration and deceleration, the centrifugal and Euler forces can transfer the liquid from the sample chamber to the collection chamber. The liquid sequential release mechanism based on the Euler force was analyzed, which showed that the angular acceleration is key to the liquid release. Then, the geometrical parameters affecting the angular acceleration of complete release were investigated and simulated. Finally, based on the relationship between the geometrical parameters of the connecting channels and the angular acceleration of complete release, a simple and precise sequential release structure was designed, which allowed for a sequential and stable transfer of the liquid into the reaction chamber. The results showed that the proposed method is capable of transferring liquid, and its simple structure, low manufacturing cost, and ease of operation enable precise sequential liquid release in centrifugal microfluidic platforms.

Conductive Adsorbent for Electrically Controlled Desorption and Sequential Release of Aqueous Mixture

Conductive Adsorbent for Electrically Controlled Desorption and Sequential Release of Aqueous Mixture

A poly (L-glutamic acid)-based porous scaffold with controllable and sequential release of kartogenin for enhanced cartilage regeneration

The controllable and sequential release of kartogenin (KGN) could improve in situ cartilage regeneration due to the intrinsically different windows between endogenous stem/progenitor cells (ESPCs) recruitment and chondrogenesis. In this paper, KGN were loaded into polyamino acid-based scaffolds in two different ways: First, phenylboronic acid ester (PBE) covalent grafting between KGN and poly (L-glutamic acid) (PLG) was built for a responsive release in an inflammatory environment, showing a responsive release of 50 % in 3 days and effectively promoting the migration of bone marrow stem cells (BMSCs) in vitro. Secondly, the conformation shift of silk fibroin (SF), from hydrophilic α-helix to hydrophobic β-sheet, could form physical network for mechanical improvement of hydrogels, as well as embedding KGN for a sustained release of 73 % in 30 days, which promoted chondrogenesis of BMSCs. In vivo rat models proved scaffolds with ROS-responsive release and physical diffusion of KGN could successfully reconstruct hyaline cartilage whose matrix composition, distribution and structure were all consistent with healthy hyaline cartilage. These results proved that this work built a smart scaffold platform with controllable and sequential release of KGN, providing a new idea for cell-free strategy in cartilage regeneration and has great clinic meaning.

Development of multi-layered three-dimensional printed scaffold for sequential delivery of biomolecules

Spatiotemporal control of exogenous growth factors plays a crucial role in the sequential repair of damaged tissues. However, traditional therapeutic trials have mostly relied on the delivery of a single molecule because of the limitations of rapid diffusion and the instability of biomolecules. In this study, we developed a novel strategy using a composite of gelatin and silica as an alternative for the stable and sequential release of biomolecules. Two biomolecules, basic fibroblast growth factor and bone morphogenetic protein-2, were incorporated into two separate composites and sequentially layered onto the activated polymer surface. Each molecule was sequentially released from each layer of the scaffold and the silica composite prevented rapid diffusion owing to its nano-porous structure. The adhesion, proliferation, and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells were significantly enhanced in the double-layered group containing separately delivered dual molecules compared with the control or single groups. Our developed gelatin–silica composite was successfully placed in double layers on polymer scaffolds, and cellular responses were promoted in this manner. These results demonstrated that our scaffolding system has potential as a therapeutic strategy for the delivery of dual or multiple biomolecules in regenerative medicine.  

Development of a sequential release nanomaterial for co-delivery of hypoxia-induced tirapazamine and HIF-1α siRNA in cancer therapy

Unlike traditional drug carriers, sequential drug delivery systems can release different drugs in order, with the first released drug providing a prerequisite for the later released drug to maximize its function, thereby achieving stronger anti-tumor effects. Herein we constructed a temporal sequential system designated TPZ@MSN/HIF-1α siRNA@PDA@GOx (MTRPG) in which mesoporous silica nanoparticles were used as cores to load hypoxia induced chemotherapy drug tirapazamine (TPZ) and gene targeted nucleic acid drug HIF-1α siRNA, polydopamine (PDA) as acid –responsive coating as well as to realize photothermal therapy, and glucose oxidase (GOx) as the outermost layer to achieve starvation therapy and construct a deepened hypoxia to activate TPZ. Through in vitro and in vivo experiments, we demonstrated that the first released glucose oxidase catalyzed the oxidation of glucose, achieving starvation treatment while reducing the acidic environment and further exacerbating hypoxia in tumor cells. The reduced acidic conditions enabled the degradation of PDA, resulting in the release of loaded HIF-1α siRNA and TPZ. At the same time, PDA could also exert photothermal therapy under 808 nm near-infrared (808 nm NIR) laser irradiation. The later released hypoxia induced chemotherapy drug TPZ amplifies its anti-tumor activity under intensified hypoxia conditions. Meanwhile, the released HIF-1α siRNA interfered with the up-regulated HIF-1α induced by the deepened hypoxia condition, which caused hypoxia tolerance in tumors, reduced its expression activity, and achieved synergistic killing of tumor cells with chemotherapy. This work provides an effective multimodal synergistic therapy strategy to promote tumor therapeutic index, which may possess a promising future in clinical application.

Stimuli-responsive gelatin-coated alginate nanocarriers: Targeted delivery of efflux pump inhibitor and antibacterial agents to control multidrug resistant P. aeruginosa

Multidrug resistance is an inescapable obstacle in healthcare settings. As the over-expression of efflux pumps is an important mediator of bacterial drug resistance, stimuli-responsive co-delivery nanosystems were developed for the sequential release of efflux pump inhibitors (EPI) and antibacterial agents. Gelatin-coated alginate nanocarriers (AlNCs@Gel) were developed containing ciprofloxacin (Cip) and quercetin (Que) as antibacterial/antibiofilm agents and EPI, respectively. Heat plot maps revealed a synergistic interaction of Cip with Que as the minimum inhibitory concentration value of Cip was reduced in the presence of EPI (quercetin) against P. aeruginosa. The encapsulation efficiency of 82 ± 1 % was observed for Cip while it was 70 ± 1 % for Que in Cip/AlNCs@Que/Gel nanocarriers. FTIR analyses confirmed an interaction between alginate and gelatin by the occurrence of a blue shift that is associated with electrostatic interactions between alginate and gelatin. These nanocarriers loaded with quercetin as EPI would release it preferentially in the presence of gelatinase enzyme and subsequently prevent the efflux of ciprofloxacin from bacterial cells. Membrane-permeability assay confirmed no permeabilization while viability studies exhibited superior bacterial inhibition, as Cip/EPI combination encapsulated in AlNCs@Gel effectively controlled the growth of P. aeruginosa as compared to antibiotic alone in AlNCs@Gel and free drug combinations. In vitro, cytotoxicity-analysis confirmed the biocompatibility of the developed core-shell nanocarriers. Furthermore, gelatin coating improved the physico-chemical properties of nanocarriers and the on-demand-release of EPI and antibiotic molecules. Thus, smartly designed AlNCs@Gel nanocarriers for the co-delivery of EPI and antibiotics present an innovative solution for preventing drug efflux from P. aeruginosa to control resistant infections.

Exploration of 4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole thioethers as potent 15-LOX inhibitors supported by in vitro, in silico, MD simulation and DNA binding studies

Inflammation is an intricate process exacerbated by the sequential release of lipid mediators during arachidonic acid metabolism through the LOX and COX pathways. The inhibition of enzymes accountable for the arachidonic acid metabolism is attributed to a synergistic anti-inflammatory impact with a broader spectrum of activity. The present study was concerned with the search for lead 15-LOX inhibitors. A new series of alkyl/aralkyl substituted 4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole (6a-h) was designed, synthesized, and characterized to determine their 15-LOX inhibitory potential. The analogue 3-(phenethylthio)-4-tolyl-5-(p-tolyloxymethyl)-4H-1,2,4-triazole (6h) was the most potent with IC50 2.73 ± 0.15 µM while 6g, 6d, 6f, 6c exhibited excellent inhibitory activities with IC50 values between 11.39 ± 0.13 and 14.63 ± 0.17 µM. All analogues maintained > 72 % blood mononuclear cells viability at 0.25 mM concentration during MTT assay. In silico ADME profiles predicted druggability potential. The molecular docking studies disclosed binding free energies of -9.78 to -7.94 kcal/mol and His373 and His378 were majorly involved in π-interactions in all molecules. Van der Waals interactions dominated hydrogen bonding in the stabilization of ligand-receptor complexes. MD simulations and free energy calculations revealed the significance of complexes in stabilizing the biomolecular system and helped in identifying the key active site residues that contributed towards the energetics. Studies against DNA dodecamer displayed minor groove binding and intercalation, suggesting a possible role in modulating DNA-related processes. The results approve the active analogues possessing promising druglike properties and work is in progress on the synthesis of more analogues in search for leads as 15-LOX inhibitors.

Enhancing Wound Healing With Sprayable Hydrogel Releasing Multi Metallic Ions: Inspired by the Body's Endogenous Healing Mechanism.

In the pursuit of new wound care products, researchers are exploring methods to improve wound healing through exogenous wound healing products. However, diverging from this conventional approach, this work has developed an endogenous support system for wound healing, drawing inspiration from the body's innate healing mechanisms governed by the sequential release of metal ions by body at wound site to promote different stages of wound healing. This work engineers a multi-ion-releasing sprayable hydrogel system, to mimic this intricate process, representing the next evolutionary step in wound care products. It comprises Alginate (Alg) and Fibrin (Fib) hydrogel infused with Polylactic acid (PLA) polymeric microcarriers encapsulating multi (calcium, copper, and zinc) nanoparticles (Alg-Fib-PLA-nCMB). Developed sprayable Alg-Fib-PLA-nCMB hydrogel show sustained release of beneficial multi metallic ions at wound site, offering a range of advantages including enhanced cellular function, antibacterial properties, and promotion of crucial wound healing processes like cell migration, ROS mitigation, macrophage polarization, collagen deposition, and vascular regeneration. In a comparative study with a commercial product (Midstress spray), developed Alg-Fib-PLA-nCMB hydrogel demonstrates superior wound healing outcomes in a rat model, indicating its potential for next generation wound care product, addressing critical challenges and offering a promising avenue for future advancements in the wound management.

Sustainable enhancement of fly ash-based geopolymers: Impact of Alkali thermal activation and particle size on green production

Addressing the challenges posed by the slow reactivity of fly ash (FA) with alkali at ambient temperatures, this study delves into the enhancement of FA-based geopolymers through alkali thermal activation (ATA). The ATA process, conducted at 550°C for 1 h, significantly increases the availability of reactive silica and alumina, which are essential for geopolymerization. A critical focus of the research is the influence of FA particle size on ATA’s efficacy and the resultant mechanical properties of the geopolymers. Findings reveal that the ATA process facilitates the rapid dissolution of the vitreous phase in FA. This leads to a sequential release of silica and alumina, which is pivotal for the geopolymer’s matrix development. Notably, geopolymers synthesized from finely milled FA, post-ATA, demonstrate a marked increase in compressive strength, escalating from 30.51 MPa to an impressive 38.46 MPa. The study meticulously delineates geopolymerization into four distinct stages—initial dissolution, depolymerization, geopolycondensation and gelation, and final diffusion, with the initial dissolution and final diffusion stages being paramount in defining the reaction kinetics and the ultimate strength of the geopolymer. This enhanced understanding paves the way for optimizing FA utilization in geopolymers, promising a more sustainable and efficient pathway for producing construction materials with superior mechanical properties.

ZIF-8-modified hydrogel sequentially delivers angiogenic and osteogenic growth factors to accelerate vascularized bone regeneration

To realize high-quality vascularized bone regeneration, we developed a multifunctional hydrogel (SHPP-ZB) by incorporating BMP-2@ZIF-8/PEG-NH2 nanoparticles (NPs) into a sodium alginate/hydroxyapatite/polyvinyl alcohol hydrogel loaded with PDGF-BB, allowing for the sequential release of angiogenic and osteogenic growth factors (GFs) during bone repair. ZIF-8 served as a protective host for BMP-2 from degradation, ensuring high encapsulation efficiency and long-term bioactivity. The SHPP-ZB hydrogel exhibited enhanced mechanical strength and injectability, making it suitable for complex bone defects. It provided a swelling interface for tissue interlocking and the early release of Zn2+ and tannin acid (TA) to exert antioxidant and antibacterial effects, followed by the sequential release of angiogenic and osteogenic GFs to promote high-quality vascularized bone regeneration. In vitro experiments demonstrated the superior angiogenic and osteogenic properties of SHPP-ZB compared to other groups. In vivo experiments indicated that the sequential delivery of GFs via SHPP-ZB hydrogel could improve vascularized bone regeneration. Further, RNA sequencing analysis of regenerative bone tissue revealed that SHPP-ZB hydrogel promoted vascularized bone regeneration by regulating JUN, MAPK, Wnt, and calcium signaling pathways in vivo. This study presented a promising approach for efficient vascularized bone regeneration in large-scale bone defects.

Purification and characterization of a thermostable Galium aparine β-galactosidase: A competent agent with enhanced cytotoxic activity against MCF-7 cell line

β-galactosidases (EC 3.2.1.23) are capable of catalysing two distinct types of reactions: transgalactosylation and hydrolysis. It is very important for the senescence of cancer cells. It is a lysosomal exoglycosidase involved in the catabolism of glycoconjugates by sequential release of β-linked terminal galactosyl residues. It has profound significance in cancer cell senescence. It can be derived from various sources including plants, animals, bacteria, yeasts, and filamentous fungi. Purification and thermodynamic characterization of β-galactosidase from Galium aparine seeds, and in vitro efficacy assessment of a breast cancer cell line. The enzyme was purified by using ammonium sulphate precipitation, dialysis, DEAE cellulose, and Sephadex G-100 chromatography. The enzyme was purified to 6.4 fold with a yield of 14 % and specific activity of 11.6 U/mg of protein. SDS-PAGE and MALDI-TOF analysis revealed that β-galactosidase is a monomer with a molecular weight of 35 kDa. The optimum pH and temperature for the purified enzyme were 6.0 and 50 ºC. The enzyme had Km and Vmax values of 0.25 mM and 32 µmol/min, respectively. The thermodynamic parameters (ΔHº, ΔGº, ΔSº) for the irreversible denaturation of the enzyme were also investigated at different temperatures (50–75 ºC). Lactose in milk was successfully hydrolyzed using a purified enzyme at a rate of 0.0121 min−1 and 12.12 min. This is better than other studied β-galactosidases. These results suggest the use of β-galactosidase from G. aparine for producing delactosed milk for the lactose-intolerant population and prebiotic synthesis. pH and optimum temperature and its activation by Ca2+ shows that it is suitable for milk processing. As a result, the enzyme was stable even at higher temperatures and had the potential to undergo catalysis, making it economically viable, primarily in the food and pharmaceutical industries. The present study is aimed to purification and thermodynamic characterization of enzyme to assess in vitro efficacy of β-galactosidase on MCF-7 cell line to delineate its therapeutic efficacy.

Biodegradable magnesium and zinc composite microspheres with synergistic osteogenic effect for enhanced bone regeneration

Biodegradable polymer microspheres in bone tissue engineering have become appealing as their non-invasive advantages in irregular damage bone repair. However, current microspheres used in BTE still lack sufficient osteogenic capacity to induce effective bone regeneration. In this study, we developed osteogenic composite microspheres concurrently loaded with magnesium oxide (MgO) and zinc oxide (ZnO), both of which are osteogenic active substances, using a facile and scalable emulsification method. The osteogenic composite microspheres exhibited a sequential yet complementary release profile characterized by a rapid release of Mg2+ and a gradual release of Zn2+ in a physiological environment, thereby maintaining the concentration of bioactive ions at a sustained high level. As a result, the combination of Mg2+ and Zn2+ in the composite microspheres led to a synergistic enhancement in biomimetic mineralization and the upregulation in the expression of osteogenic-related genes and proteins at the cellular level. Through a critical-sized calvarial rate defect model, the osteogenic composite microspheres were demonstrated to have strong osteogenic ability to promote new bone formation via ultrasonic imaging, histological and immunohistochemical evaluations. In sum, these osteogenic composite microspheres as microcarriers of Mg2+ and Zn2+ have great potential in the delivery of therapeutic ions for treating bone defects.

Ultrasound-Activatable In Situ Vaccine for Enhanced Antigen Self- and Cross-Presentation to Overcome Cancer Immunotherapy Resistance.

Insufficient antigen self-presentation of tumor cells and ineffective antigen cross-presentation by dendritic cells (DCs) contribute to diminished immune recognition and activation, which cause resistance to immunotherapies. Herein, we present an ultrasound-activatable in situ vaccine by utilizing a hybrid nanovesicle composed of a thylakoid (TK)/platelet (PLT) membrane and a liposome encapsulating DNA methyltransferase inhibitor zebularine (Zeb) and sonosensitizer hematoporphyrin monomethyl ether (HMME). Upon local exposure to ultrasound, reactive oxygen species (ROS) are generated and induce the sequential release of the payloads. Zeb can efficiently inhibit tumor DNA hypermethylation, promoting major histocompatibility complex class I (MHC-I) molecules-mediated antigen self-presentation to improve immune recognition. Meanwhile, the catalase on the TK membrane can decompose the tumoral overexpressed H2O2 into O2, which boosts the generation of ROS and the destruction of tumor cells, resulting in the in situ antigen release and cross-presentation of tumor antigens by DCs. This in situ vaccine simultaneously promotes antigen self-presentation and cross-presentation, resulting in heightened antitumor immunity to overcome resistance.

Clinic-oriented injectable smart material for the treatment of diabetic wounds: Coordinating the release of GM-CSF and VEGF

Chronic wounds are often caused by diabetes and present a challenging clinical problem due to vascular problems leading to ischemia. This inhibits proper wound healing by delaying inflammatory responses and angiogenesis. To address this problem, we have developed injectable particle-loaded hydrogels which sequentially release Granulocyte-macrophage- colony-stimulating-factor (GM-CSF) and Vascular endothelial growth factor (VEGF) encapsulated in polycaprolactone-lecithin-geleol mono-diglyceride hybrid particles. GM-CSF promotes inflammation, while VEGF facilitates angiogenesis. The hybrid particles (200–1000 nm) designed within the scope of the study can encapsulate the model proteins Bovine Serum Albumin 65 ± 5 % and Lysozyme 77 ± 10 % and can release stably for 21 days. In vivo tests and histological findings revealed that in the hydrogels containing GM-CSF/VEGF-loaded hybrid particles, wound depth decreased, inflammation phase increased, and fibrotic scar tissue decreased, while mature granulation tissue was formed on day 10. These findings confirm that the hybrid particles first initiate the inflammation phase by delivering GM-CSF, followed by VEGF, increasing the number of vascularization and thus increasing the healing rate of wounds. We emphasize the importance of multi-component and sequential release in wound healing and propose a unifying therapeutic strategy to sequentially deliver ligands targeting wound healing stages, which is very important in the treatment of the diabetic wounds.