Process optimization for microfluidic preparation of liposomes using food-grade components.

This study explores the potential for optimizing a sustainable manufacturing process that maintains the essential characteristics of conventional liposomes using food-grade solvents and components. The focus was comparing the physicochemical, morphological, and interfacial properties of liposomes produced with these food-grade ingredients to those made by conventional methods. It was found that there was no significant difference in particle size (195.87 ± 1.40 nm) and ζ-potential (-45.13 ± 0.65 mV) between liposomes made from food-grade and conventional materials. The manufacturing process for liposomes, utilizing food-grade solvents and components, was optimized through the application of Plackett-Burman design and response surface methodology. This approach helped identify key parameters (soy lecithin, ß-sitosterol, W/O ratio) and their optimal values (3.17 g, 0.25 g, 1:2.59). These findings suggest that it is possible to enhance the use of liposomes as an effective and safe delivery system in the food industry, adhering to the strict guidelines set by regulatory agencies.

Natural-Wood-Inspired Ultrastrong Anisotropic Hybrid Hydrogels Targeting Artificial Tendons or Ligaments.

Hydrogels are able to mimic the flexibility of biological tissues or skin, but they still cannot achieve satisfactory strength and toughness, greatly limiting their scope of application. Natural wood can offer inspiration for designing high-strength hydrogels attributed to its anisotropic structure. Herein, we propose an integrated strategy for efficient preparation of ultrastrong hydrogels using a salting-assisted prestretching treatment. The as-prepared poly(vinyl alcohol)/cellulose nanofiber hybrid hydrogels show distinct wood-like anisotropy, including oriented molecular fiber bundles and extended grain size, which endows materials with extraordinarily comprehensive mechanical properties of ultimate breaking strength exceeding 40 MPa, strain approaching 250%, and toughness exceeding 60 MJ·m-3, and outstanding tear resistance. Impressively, the breaking strength and toughness of the reswollen preoriented hydrogels approach 10 MPa and 25 MJ·m-3, respectively. In vitro and in vivo tests demonstrate that the reswollen hydrogels do not affect the growth and viability of the cells, nor do they cause the inflammation or rejection of the mouse tissue, implying extremely low biotoxicity and perfect histocompatibility, showcasing bright prospects for application in artificial ligaments or tendons. The strategy provided in this study can be generalized to a variety of biocompatible polymers for the fabrication of high-performance hydrogels with anisotropic structures.

Stepwise Size Shrinkage Cascade-Activated Supramolecular Prodrug Boosts Antitumor Immunity by Eliciting Pyroptosis.

Effective pyroptosis induction is a promising approach to potentiate cancer immunotherapy. However, the actual efficacy of the present pyroptosis inducers can be weakened by successive biological barriers. Here, a cascaded pH-activated supramolecular nanoprodrug (PDNP) with a stepwise size shrinkage property is developed as a pyroptosis inducer to boost antitumor immune response. PDNPs comprise multiple poly(ethylene glycol) (PEG) and doxorubicin (DOX) drug-polymer hybrid repeating blocks conjugated by ultra-pH-sensitive benzoic imine (bzi) and hydrazone (hyd) bonds. The PEG units endow its "stealth" property and ensure sufficient tumor accumulation. A sharp switch in particle size and detachment of PEG shielding can be triggered by the acidic extracellular pH to achieve deep intratumor penetration. Following endocytosis, second-stage size switching can be initiated by more acidic endolysosomes, and PDNPs disassociate into ultrasmall cargo to ensure accurate intracellular delivery. The cascaded pH activation of PDNPs can effectively elicit gasdermin E (GSDME)-mediated pyroptosis to enhance the immunological response. In combination with anti-PD-1 antibody, PDNPs can amplify tumor suppression and extend the survival of mice, which suggests a powerful immune adjuvant and pave the way for high-efficiency immune checkpoint blockade therapy.

A 3D bioprinted decellularized extracellular matrix/gelatin/quaternized chitosan scaffold assembling with poly(ionic liquid)s for skin tissue engineering.

Currently, a suitable bioink for 3D bioprinting and capable of mimicking the microenvironment of native skin and preventing bacterial infection remains a major challenge in skin tissue engineering. In this study, we prepared a tissue-specific extracellular matrix-based bioink, and dECM/Gel/QCS (dGQ) 3D scaffold assembling with poly(ionic liquid)s (PILs) (dGQP) was obtained by an extrusion 3D bioprinting technology and dynamic hydrogen bonding method. The morphologies, mechanical properties, porosity, hydrophilicity, biodegradation, hemostatic effect, antibacterial ability, and biocompatibility of the hybrid scaffolds were characterized and evaluated. Results showed that the rapid release (2 h) of PILs on the dGQP scaffold can quickly kill gram-negative (E. coli) and gram-positive (S. aureus) bacteria with almost 100 % antibacterial activity and maintained a stable sterile environment for a long time (7 d), which was superior to the dGQ scaffold. The hemostasis and hemolysis test showed that the dGQP scaffold had a good hemostatic effect and excellent hemocompatibility. In vitro cytocompatibility studies showed that although the cell growth on dGQP scaffold was slow in the early stage, the cells proliferated rapidly since day 4 and had high ECM secretion at day 7. Overall, this advanced dGQP scaffold has a considerable potential to be applied in skin tissue engineering.

Promoting lignocellulose degradation during green waste composting by maintaining a specific temperature through heap size control.

Owing to its high lignocellulose content, the recalcitrance of green waste is a technical challenge obstructing the composting process. This study aimed to identify a temperature that could facilitate efficient lignin and cellulose degradation during green waste composting, and maintain this temperature by controlling the heap size to enhance the degradation. The optimum temperature was determined by conducting a laboratory-scale cultivation experiment under controlled temperatures, and a pilot-scale experiment was conducted to explore heap size control and its influence on green waste composting. The results showed that efficient lignin and cellulose degradation was achieved when the temperature was between 45 and 60 ℃, and maintaining this temperature for at least 150 days maximized the lignin and cellulose degradation rates. This was achieved by constraining the heap size at 0.8 m3 at the beginning of composting; 1.56, 2.60, and 4.00 m3 on days 15, 39, and 96; and then enlarging the heap as much as possible on day 156. Following this approach, the duration of the target temperature was extended by over six times, the lignin and cellulose degradation rates were increased by 18.82-21.38 % and 9.54-11.55 %, and nitrification and humification were enhanced. Correlation analysis showed that lignocellulose degradation, nitrification, and humification were positively and significantly correlated with the duration of the target temperature. Generally, heap size control is an ecological and economic method of enhancing the efficiency and quality of green waste composting and compost, respectively.

Application of quadratic regression orthogonal design to develop a composite inoculum for promoting lignocellulose degradation during green waste composting.

The aims of this study are to determine the feasibility of applying QROD (quadratic regression orthogonal design) to optimize a combination of microorganisms and to develop a composite inoculum for promoting lignocellulose degradation during GWC (green waste composting). This feasibility was studied in a laboratory scale experiment, using three lignocellulolytic microorganisms, isolated from the mature phase of GWC by the dilution plating method. After the feasibility was confirmed, a composite inoculum was developed through the results of the optimization, whose effect was evaluated by comparing it with Phanerochaete chrysosporium and EM (Effective Microorganisms) in a pilot scale experiment of GWC. The use of QROD to finish this optimization was proven feasible, because the p value of the regression equation was less than 0.05 (0.0108), meaning that the quadratic regression model is suitable for describing the relationship between the combination of the three microorganisms and their ability to degrade lignocellulose. Additional proof of this feasibility is that the composite inoculum in the quadratic regression orthogonal experiment demonstrated lignocellulose degradation ability similar to the GWC experiment. Although the lignin degradation ability of the composite inoculum did not surpass Phanerochaete chrysosporium, it was stronger than EM. Meanwhile, cellulose degradation ability and humus synthesis ability of the composite inoculum were stronger than for Phanerochaete chrysosporium and were close to EM. It is hard to tell which inoculum is the best since each inoculum had advantages in different aspects, while the composite inoculum still showed a considerable effect of lignocellulose degradation during GWC.

[EXPERIMENTAL STUDY ON CHITOSAN/ALLOGENEIC BONE POWDER COMPOSITE POROUS SCAFFOLD TO REPAIR BONE DEFECTS IN RATS].

OBJECTIVE: To explore the feasibility of chitosan/allogeneic bone powder composite porous scaffold as scaffold material of bone tissue engineering in repairing bone defect. METHODS: The composite porous scaffolds were prepared with chitosan and decalcified allogeneic bone powder at a ratio of 1 : 5 by vacuum freeze-drying technique. Chitosan scaffold served as control. Ethanol alternative method was used to measure its porosity, and scanning electron microscopy (SEM) to measure pore size. The hole of 3.5 mm in diameter was made on the bilateral femoral condyles of 40 adult Sprague Dawley rats. The composite porous scaffolds and chitosan scaffolds were implanted into the hole of the left femoral condyle (experimental group) and the hole of the right femoral condyle (control group), respectively. At 2, 4, 8, and 12 weeks after implantation, the tissues were harvested for gross observation, histological observation, and immunohistochemical staining. RESULTS: The composite porous scaffold prepared by vacuum freeze-drying technique had yellowish color, and brittle and easily broken texture; pore size was mostly 200-300 µm; and the porosity was 76.8% ± 1.1%, showing no significant difference when compared with the porosity of pure chitosan scaffold (78.4% ± 1.4%) (t = -2.10, P = 0.09). The gross observation and histological observation showed that the defect area was filled with new bone with time, and new bone of the experimental group was significantly more than that of the control group. At 4, 8, and 12 weeks after implantation, the bone forming area of the experimental group was significantly larger than that of the control group (P < 0.05). The immunohistochemical staining results showed that osteoprotegerin (OPG) positive expression was found in the experimental group at different time points, and the positive expression level was significantly higher than that in the control group (P < 0.05). CONCLUSION: Chitosan/allogeneic bone powder composite porous scaffold has suitable porosity and good osteogenic activity, so it is a good material for repairing bone defect, and its bone forming volume and bone formation rate are better than those of pure chitosan scaffold.

Folded Elastic Strip-Based Triboelectric Nanogenerator for Harvesting Human Motion Energy for Multiple Applications.

A folded elastic strip-based triboelectric nanogenerator (FS-TENG) made from two folded double-layer elastic strips of Al/PET and PTFE/PET can achieve multiple functions by low frequency mechanical motion. A single FS-TENG with strip width of 3 cm and length of 27 cm can generate a maximum output current, open-circuit voltage, and peak power of 55 µA, 840 V, and 7.33 mW at deformation frequency of 4 Hz with amplitude of 2.5 cm, respectively. This FS-TENG can work as a weight sensor due to its good elasticity. An integrated generator assembled by four FS-TENGs (IFS-TENG) can harvest the energy of human motion like flapping hands and walking steps. In addition, the IFS-TENG combined with electromagnetically induced electricity can achieve a completely self-driven doorbell with flashing lights. Moreover, a box-like generator integrated by four IFS-TENGs inside can work in horizontal or random motion modes and can be improved to harvest energy in all directions. This work promotes the research of completely self-driven systems and energy harvesting of human motion for applications in our daily life.

[Comparative study on repairing rabbit radius segmental defects with two different proportions of chitosan combined with allogeneic morselized bone].

OBJECTIVE: To give a preliminary experimental evidence and to prove chitosan and allogeneic morselized bone as potential bone substitutions in repairing rabbit radius segmental defect. METHODS: Chitosan and allogeneic morselized bone were mixed with various ratios (1:5, 1:10, 1:25, 1:50, and 1:100). After preparation, the physical and chemical properties of the composites were preliminary detected; the composites at the ratios of 1:50 and 1:25 had good physical and chemical properties and were used for the animal experiment. The radius segmental defects of 15 mm in length were made in 50 adult New Zealand white rabbits (weighing 2.5-3.0 kg), then the animals were divided into 2 groups. In groups A and B, chitosan/allogeneic morselized bone composites were implanted at the ratio of 1:50 and 1:25, respectively. After 1, 2, 4, 8, and 12 weeks of operation, the gross, histological, immunohistochemical observations were performed. Before the rabbits were sacrificed, X-ray films were taken; the serum calcium and alkaline phosphatase (ALP) concentration were measured; and the biomechanical measurement was carried out at 12 weeks. RESULTS: The results of gross observation were essentially consistent with those of the X-ray films. The histological observation showed that the bone formation was earlier in group A than in group B; the amount of new bone formation in group A was more than that in group B; and the bone forming area in group A was bigger than that in group B (P < 0.05) at 4 and 8 weeks after operation. The immunohistochemical staining showed that vascular endothelial growth factor and insulin-like growth factor receptor II proteins expressed in the cytoplasm of 2 groups after 4 and 8 weeks, and the expression in group A was higher than that in group B (P < 0.05). There was no significant difference in the serum calcium concentration between 2 groups at each time point (P > 0.05). After 4 and 8 weeks, the ALP concentration in group A was significantly higher than that in group B (P < 0.05). After 12 weeks, the radius maximum bending loads of groups A and B were (299.75 +/- 27.69) N and (278.54 +/- 17.09) N, respectively, showing significant difference (t=4.045, P=0.002). CONCLUSION: The composite of chitosan and allogeneic morselized bone has good osteogenic activity and can be used as a bone tissue engineering scaffold, and the optimum ratio of chitosan to allogeneic morselized bone was 1:50.

[A preliminary study of high viscous chitosan/glycerol phosphate with demineralized bone matrix to repair cartilage defects in rabbits].

OBJECTIVE: To evaluate the effect of implantation of the complex of high viscous chitosan/glycerol phosphate with demineralized bone matrix (HV-C/GP-DBM) in repairing cartilage defects of rabbits. METHODS: HV-C/GP-DBM was prepared by compounding HV-C/GP and DBM by 2:1 (W/W). Twenty-four 34-week-old New Zealand white adult rabbits, weighing 3.5-4.5 kg, were included. A bit with the diameter of 3.5 mm was used to drill 3-cm-deep holes in both sides of femoral condyle to make cartilage defects. The complex of HV-C/GP-DBM was then injected into the right holes as the experimental group and the left ones serve as the control group. The rabbits were killed at 4, 8 and 16 weeks after the operation, respectively. The obtained specimens were observed macroscopically, microscopically and histologically. According to the International Cartilage Repair Society Histological Scoring (ICRS), the effect of cartilage repair was assessed at 16 weeks postoperatively. RESULTS: At 4-8 weeks postoperatively, in the experimental group, the defects were filled with hyaline cartilage-like tissues; the majority of chitosan degradated; and the DBM particles were partly absorbed. However, in the control group, there were small quantities of discontinuous fibrous tissues and maldistributed chondrocytes at the border and the bottom of the defects. At 16 weeks postoperatively, 6 joints in the experimental group had smooth surface, and the defects were basically repaired by hyaline cartilage-like tissues. The newly-formed tissues integrated well with the surrounding area. Under the cartilage, the new bone formation was still active and some DBM particles could be seen. However, the defects in the control group were repaired by fibrous tissues. The result of histological scoring of the specimens at 16 weeks showed that a total of 6 aspects including formation of chondrocytes and integration with the surrounding cartilages were superior in the experimental group to those in the control group, and there were significant differences between the two groups (P < 0.05). CONCLUSION: The biodegradable and injectable complex of HV-C/GP-DBM with good histocompatibility and non-toxic side effects can repair cartilage defects and is a promising biomaterial for cartilage defect repair.