Macrophage membrane modified baicalin liposomes improve brain targeting for alleviating cerebral ischemia reperfusion injury.

Baicalin (BA) has a good intervention effect on encephalopathy. In this study, macrophage membrane was modified on the surface of baicalin liposomes (BA-LP) by extrusion method. Macrophage membrane modified BA-LP (MM-BA-LP) was characterized by various analytical techniques, and evaluated for brain targeting. The results presented MM-BA-LP had better brain targeting compared with BA-LP. Pharmacokinetic experiments showed that MM-BA-LP improved pharmacokinetic parameters and increased the residence time of BA. Pharmacodynamic of middle cerebral artery occlusion (MCAO) rat model was studied to verify the therapeutic effect of MM-BA-LP on cerebral ischemia reperfusion injury (CIRI). The results showed that MM-BA-LP could significantly improve the neurological deficit, cerebral infarction volume and brain pathological state of MCAO rats compared with BA-LP. These results suggested that MM-BA-LP could significantly enhance the brain targeting and improve the circulation of BA in blood, and had a significantly better neuroprotective effect on MCAO rats than BA-LP.

Geniposide-Loaded Liposomes for Brain Targeting: Development, Evaluation, and In Vivo Studies.

Geniposide (GE) possesses excellent neuroprotective effects but with poor brain targeting and short half-life. Liposome was considered to have great potential for brain diseases. Therefore, this research aimed to develop a geniposide liposome (GE-LP) as a brain delivery system for cerebral ischemia reperfusion injury (CIRI) therapy and evaluate its characterization, pharmacokinetics, brain targeting, and neuroprotective effects in vivo. Then, a reverse-phase evaporation method was applied to develop the GE-LP and optimize the formulation. Notably, the GE-LP had suitable size, which was 223.8 nm. Subsequently, the pharmacokinetic behavior of GE solution and GE-LP in mice plasma was investigated, and the brain targeting was also researched. The results showed that GE in plasma of GE-LP displayed three folds longer distribution half-life and a higher bioavailability and brain targeting compared to GE solution. In vivo neuroprotective effects was evaluated through the middle cerebral artery occlusion (MCAO) rat model, and GE-LP exhibited a stronger tendency in preventing the injury of CIRI, which can significantly improve neurological deficits. Overall, this study demonstrates GE-LP as a new formulation with ease of preparation, sustained release, and high brain targeting, which has significant development prospects on CIRI; this is expected to improve the efficacy of GE and reduce the frequency of administration.

Baicalin Liposome Alleviates Lipopolysaccharide-Induced Acute Lung Injury in Mice via Inhibiting TLR4/JNK/ERK/NF-κB Pathway.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are challenging diseases with the high mortality in a clinical setting. Baicalin (BA) is the main effective constituent isolated from the Chinese medical herb Scutellaria baicalensis Georgi, and studies have proved that it has a protective effect on ALI induced by lipopolysaccharide (LPS) due to the anti-inflammatory efficacy. However, BA has low solubility which may limit its clinical application. Hence, we prepared a novel drug delivery system-Baicalin liposome (BA-LP) in previous research-which can improve some physical properties of BA. Therefore, we aimed to explore the effect of BA-LP on ALI mice induced by LPS. In pharmacokinetics study, the values of t 1/2 and AUC0- t in the BA-LP group were significantly higher than that of the BA group in normal mice, indicating that BA-LP could prolong the duration time in vivo of BA. The BA-LP group also showed a higher concentration in lung tissues than the BA group. Pharmacodynamics studies showed that BA-LP had a better effect than the BA group at the same dosage on reducing the W/D ratio, alleviating the lung injury score, and decreasing the proinflammatory factors (TNF-α, IL-1ß) and total proteins in bronchoalveolar lavage fluids (BALF). In addition, the therapeutic effects of BA-LP showed a dose-dependent manner. Western blot analysis indicated that the anti-inflammatory action of BA could be attributed to the inhibition of the TLR4-NFκBp65 and JNK-ERK signaling pathways. These results suggest that BA-LP could be a valuable therapeutic candidate in the treatment of ALI.

Review on vat photopolymerization additive manufacturing of bioactive ceramic bone scaffolds.

Bone defects frequently occur in clinical settings due to trauma, disease, tumors, and other causes. The clinical use of autologous bones and allograft bone, however, has several limitations, such as limited sources, donor site morbidity, and immunological rejection. Nevertheless, there is newfound hope for regenerating and repairing bone defects through the development and integration of bone tissue engineering scaffold and additive manufacturing (AM) technology, also known as 3D printing. In particular, vat photopolymerization (VPP)-AM of bioactive ceramic bone scaffolds has garnered significant interest from interdisciplinary researchers in recent years. On the one hand, VPP-AM demonstrates clear advantages in printing accuracy and speed compared to other AM and non-AM technologies. On the other hand, bioactive ceramic materials exhibit superior bioactivity, biodegradability, and mechanical properties compared to metals, polymers, and bioinert ceramics, making them one of the most promising biomaterials for developing bone scaffolds. This paper reviews the research progress of VPP-AM of bioactive ceramic bone scaffolds, covering the process principles of various VPP-AM technologies, the performance requirements and preparation process of VPP ceramic slurry, the VPP process of bioactive ceramic bone scaffolds, and the research progress on different material types of VPP bioactive ceramic scaffolds. Firstly, we provide a brief introduction to the process principles and medical applications of various VPP technologies. Secondly, we explore the composition of the VPP ceramic slurry system, discussing the function of various components and their effects on printing quality. Thirdly, we delve into the performance requirements of bone scaffolds and summarize the research progress of VPP bioactive ceramic bone scaffolds of various material types including hydroxyapatite (HA), tricalcium phosphate (TCP), bioglass (BG), etc.; Finally, we discuss the challenges currently faced by VPP-AM bioactive ceramic bone scaffolds and propose possible development directions for the future.

Brain targeted borneol-baicalin liposome improves blood-brain barrier integrity after cerebral ischemia-reperfusion injury via inhibiting HIF-1α/VEGF/eNOS/NO signal pathway.

Baicalin (BA) is widely used in the treatment of cerebral ischemia-reperfusion injury (CIRI). The key to treating encephalopathy is to increase the amounts of drugs entering the brain. Borneol-baicalin liposome (BO-BA-LP) was prepared in previous research based on the characteristics of borneol (BO) in promoting drug brain entry. In this study, the effect of BO-BA-LP on improving blood-brain barrier (BBB) integrity was researched. Results showed BO-BA-LP may increase ability of BA to penetrate the cell membrane in vitro. Pharmacokinetic results showed the BO-BA-LP could increase concentrations of BA in plasma and brain tissues of normal and CIRI mice. Pharmacological results revealed BO-BA-LP could improve the neurological function, brain edema, and histopathology of CIRI mice. Besides, BO-BA-LP could protect BBB by regulating hypoxia inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF)/endothelial nitric oxide synthase (eNOS)/nitric oxide (NO) pathway. The research showed that BO in BO-BA-LP could increase the absorption of BA by increasing BBB permeability, leading to a better therapeutic effect of BO-BA-LP on CIRI mice.

3D printed TPMS structural PLA/GO scaffold: Process parameter optimization, porous structure, mechanical and biological properties.

Bone scaffolds should have good biocompatibility and mechanical and biological properties, which are primarily by the material design, porous structure, and preparation process. In this study, we proposed polylactic acid (PLA) as the base material, graphene oxide (GO) as an enhancing filler, triply periodic minimal surface (TPMS) as a porous structure, and fused deposition modeling (FDM) 3D printing as a preparation technology to develop a TPMS structural PLA/GO scaffold and evaluate their porous structures, mechanical properties, and biological properties towards bone tissue engineering. Firstly, the influence of the FDM 3D printing process parameters on the forming quality and mechanical properties of PLA was studied by orthogonal experimental design, based on which the process parameters were optimized. Then, GO was composited with PLA, and PLA/GO nanocomposites were prepared by FDM. The mechanical tests showed that GO can effectively improve the tensile and compression strength of PLA; only by adding 0.1% GO the tensile and compression modulus was increased by 35.6% and 35.8%, respectively. Then, TPMS structural (Schwarz-P, Gyroid) scaffold models were designed and TPMS structural PLA/0.1%GO nanocomposite scaffolds were prepared by FDM. The compression test showed that the TPMS structural scaffolds had higher compression strength than the Grid structure; This was owing to the fact that the continuous curved structure of TMPS alleviated stress concentration and had a more uniform stress bearing. Moreover, cell culture indicated bone marrow stromal cells (BMSCs) showed better adhesion, proliferation, and osteogenic differentiation behaviors on the TPMS structural scaffolds as the continuous surface structure of TPMS had better connectivity and larger specific surface area. These results suggest that the TPMS structural PLA/GO scaffold has potential application in bone repair. This article suggests the feasibility of co-designing the material, structure, and technology for achieving the good comprehensive performance of polymer bone scaffolds.

Preparation, characterization and safety evaluation of Ligusticum chuanxiong essential oils liposomes for treatment of cerebral ischemia-reperfusion injury.

The essential oils of Ligusticum chuanxiong Hort. (CXEO) are considered to be important parts of the pharmacological action of Ligusticum chuanxiong Hort. CXEO have a wide range of applications in various fields. Despite the interesting properties of CXEO, the volatility and low solubility have limited the application. Liposomes are vesicles composed of concentric bilayer lipids arranged around the water environment. Therefore, this study aimed to prepare stable CXEO liposomes (CXEO-LP) to improve the properties. Then, CXEO-LP were prepared by thin film dispersion method and optimized. The results showed that CXEO-LP were well dispersed. Subsequently, in vitro release and antioxidant properties of CXEO-LP were researched. CXEO-LP had slow release effect and oxidation resistance, indicating CXEO-LP may be a potential drug for treating cerebral ischemia-reperfusion injury (CIRI). The nasal mucosa toxicity test and acute toxicity test showed that CXEO-LP had no obvious toxicity to nasal cavity, heart, liver, spleen, lung and kidney tissues. Pharmacodynamic studies found that CXEO-LP significantly improved neurological deficits and brain pathology in a mouse model of CIRI compared to CXEO after intranasal administration. In general, this study showed that CXEO-LP were easy to prepare and continuously released, and had an important development prospect in the treatment of CIRI.

Multidrug-loaded liposomes prevent ischemic stroke through intranasal administration.

Baicalin (BA), a multi-target neuroprotective agent, has poor solubility resulting in low bioavailability. In this study, multidrug-loaded liposomes were prepared by encapsulating BA, borneol (BO) and cholic acid (CA) to prevent ischemic stroke. BBC-LP were administered intranasally (i.n.) to deliver into the brain for neuroprotection. Finally, potential mechanism of BBC treating ischemic stroke (IS) was explored by network pharmacology. In this study, BBC-LP was prepared by reverse evaporation method, and the encapsulation efficiency (EE) of the optimized liposomes was 42.69% and the drug loading (DL) was 6.17%. The liposomes had low mean particle size (156.62 ± 2.96 nm), polydispersity index (PDI) (0.195) and zeta potential (-0.99 mv). Compared to BBC, pharmacodynamic studies revealed that BBC-LP significantly improved neurological deficits, brain infarct volume, and cerebral pathology in MCAO rats. Toxicity studies showed that BBC-LP was not irritating to the nasal mucosa. These results suggest that BBC-LP can safely and effectively ameliorate IS injury by i.n. administration. Moreover, it's neuroprotective function may be related to the anti-apoptotic and anti-inflammatory effects exerted by phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway and mitogen-activated protein kinase (MAPK) signaling pathway.

A natural biomineral for enhancing the biomineralization and cell response of 3D printed polylactic acid bone scaffolds.

Polylactic acid (PLA) has been extensively used as a bone scaffold material, but it still faces many problems including low biomineralization ability, weak cell response, low mechanical properties, etc. In this study, we proposed to utilize the distinctive physical, chemical and biological properties of a natural biomineral with organic matrix, pearl powder, to enhance the overall performance of PLA bone scaffolds. Porous PLA/pearl composite bone scaffolds were prepared using fused deposition modeling (FDM) 3D printing technology, and their comprehensive performance was investigated. Macro- and micro- morphological observation by the optical camera and scanning electron microscopy (SEM) showed the 3D printed scaffolds have interconnected and ordered periodic porous structures. Phase analysis by X-ray diffraction (XRD) indicated pearl powder was well composited with PLA without impurity formation during the melt extrusion process. The mechanical test results indicated the tensile and compressive strength of PLA/pearl composite scaffolds with 10 % pearl powder content yielded the highest values, which were 15.5 % and 21.8% greater than pure PLA, respectively. The water contact angle and water absorption tests indicated that PLA/pearl showed better hydrophilicity than PLA due to the presence of polar groups in the organic matrix of the pearl powder. The results of the simulated body fluid (SBF) soaking revealed that the addition of pearl powder effectively enhanced the formation and deposition of apatite, which was attributed to the release of Ca2+ from the dissolution of pearl powder. The cell culture of bone marrow mesenchymal stem cells (BMSCs) indicated that PLA/pearl scaffolds showed better cell proliferation and osteogenic differentiation than PLA due to the stimulation of the biological organic matrix in pearl powder. These outcomes signify the potential of pearl powder as a natural biomineral containing bio-signal factors to improve the mechanical and biological properties of polymers for better bone tissue engineering application.

Transcriptional analysis reveals sodium nitroprusside affects alfalfa in response to PEG-induced osmotic stress at germination stage.

Drought is one of the most common environmental factors that affect alfalfa germination and development. Nitric oxide (NO) could mediate stress tolerance in plants. The goal of this study was to determine exogenous NO donor-mediated drought adaption molecular mechanisms during the alfalfa germination stage. In this study, physiological and transcriptome analyses were performed on 7 days of the growth period seedlings by sodium nitroprusside (SNP) and polyethylene glycol (PEG) treatment. The results showed that SNP supplementation alleviated malondialdehyde accumulation, increased levels of proline and soluble sugars, and enhanced antioxidant enzyme activity under osmotic stress conditions. RNA-Seq experiments identified 5828 genes exhibiting differential expression in seedlings treated with PEG, SNP, or SNP+PEG relative to seedlings treated with distilled water. Of these DEGs, 3235 were upregulated, and 2593 were downregulated relative to the controls. Fifteen DEGs were amplified by qRT-PCR to verify the changes in expression determined by RNA-Seq, revealing that PIF3, glnA, PLCG1, and RP-S11e exhibited enhanced expression under the SNP+PEG treatment. SNP was found to modulate redox homeostasis-related genes such as GSTs, SOD2, GPX, and RBOH, and triggered calcium signaling transduction. It also induced some key genes relating to the abscisic acid, ethylene, and auxin signaling transduction in response to PEG stress. Conversely, genes associated with secondary metabolite biosynthesis and the metabolism of starch and sucrose during osmotic stress were downregulated by SNP. These results provide new insights into SNP-mediated drought adaption mechanisms at transcriptome-wide in alfalfa and reveal key drought tolerance pathways in this species.

Pharmacokinetics, pharmacodynamics and toxicity of Baicalin liposome on cerebral ischemia reperfusion injury rats via intranasal administration.

Baicalin (BA) is a major active component from the traditional Chinese medicine, which has been widely used to treat brain diseases. Previously, the baicalin liposome (BA-LP) was prepared to improve its low bioavailability. However, the existence of the obstacles such as the blood-brian-barrier (BBB) still make it difficult to enter brain effectively. Meanwhile, many reports have shown that drugs can be transported into brain through intranasal administration without the BBB. Therefore, we aim to explore the effect of BA-LP on middle cerebral artery occlusion (MCAO) rats via i.n. administration. The results showed that BA and BA-LP had no obvious impact on mucosa after i.n. administration. And in pharmacokinetics study after i.n. administration, the value of t 1/2 and AUC 0-t in BA-LP group were significantly higher than that of the BA group (p < 0.05), indicating BA-LP could prolong the extension time of BA in vivo and further improve the bioavailability. In the brain biodistribution, the BA-LP group showed a higher BA concentration in brain tissues. Pharmacodynamics studies showed that BA-LP through i.n. administration could significantly improve the neurological deficits, cerebral infarction and brain pathological status in rats with MCAO surgery. Obviously, the BA-LP was more effective after nasal administration than intravenous administration, suggesting the nasal administration is more advantageous route in brain concentration enrichment. In conclusion, BA-LP could be safely used in i.n. administration, which can effectively improve brain targeting effect and thus protect the MCAO rats. Furthermore, successful use of the BA-LP via nasal delivery can provide a model for other drugs with neuroprotective effect and further promote the cure rate of brain diseases.

Preparation, Characterization and in vivo Study of Borneol-Baicalin-Liposomes for Treatment of Cerebral Ischemia-Reperfusion Injury.

PURPOSE: Baicalin (BA) has a good neuroprotective effect, but it is eliminated quickly in the body and does not easily reach the brain. In this experiment, borneol (BO) was used as an auxiliary drug to prepare borneol-baicalin-liposomes (BO-BA-LP) to prolong the efficacy time of BA, synergistically synergize, introduce drugs into the brain, and better exert the therapeutic effect on cerebral ischemia-reperfusion (I/R) injury. METHODS: Through single-factor inspection and response surface optimization analysis, obtained the best preparation process of BO-BA-LP and characterized by various analytical techniques. Validated the long-term effectiveness of BA-BO-LP through pharmacokinetic studies and conducted pharmacodynamic studies on the middle cerebral artery occlusion (MCAO) rat model to verify the therapeutic effect of BO-BA-LP on cerebral I/R injury. RESULTS: The optimum preparation conditions of BO-BA-LP were as follows: the dosage of BO was 9.55 mg, the ratio of phospholipid to drug was 4.02:1, the ratio of phospholipid to cholesterol was 7.25:1, the entrapment efficiency (EE) was 41.49%, and the drug loading (DL) was 4.29%. The particle size range of the liposomes was 167.1 nm, and the polydispersity index (PDI) range was 0.113. The results of pharmacokinetic experiments showed that the combination of BA and BO liposomes effectively improved the pharmacokinetic parameters of BA and prolonged the half-life of BA. Pharmacodynamic studies have found that, compared with BA-LP, BO-BA-LP can significantly improve neurological deficits, cerebral infarction volume, and brain pathological states on MCAO rats. CONCLUSION: These results demonstrated that BO-BA-LP can improve the circulation of drugs in the blood, and the addition of BO can enhance the therapeutic effect of BA and effectively improve cerebral I/R.

Identification of Exogenous Nitric Oxide-Responsive miRNAs from Alfalfa (Medicago sativa L.) under Drought Stress by High-Throughput Sequencing.

Alfalfa (Medicago sativa L.) is a high quality leguminous forage. Drought stress is one of the main factors that restrict the development of the alfalfa industry. High-throughput sequencing was used to analyze the microRNA (miRNA) profiles of alfalfa plants treated with CK (normal water), PEG (polyethylene glycol-6000; drought stress), and PEG + SNP (sodium nitroprusside; nitric oxide (NO) sprayed externally under drought stress). We identified 90 known miRNAs belonging to 46 families and predicted 177 new miRNAs. Real-time quantitative fluorescent PCR (qRT-PCR) was used to validate high-throughput expression analysis data. A total of 32 (14 known miRNAs and 18 new miRNAs) and 55 (24 known miRNAs and 31 new miRNAs) differentially expressed miRNAs were identified in PEG and PEG + SNP samples. This suggested that exogenous NO can induce more new miRNAs. The differentially expressed miRNA maturation sequences in the two treatment groups were targeted by 86 and 157 potential target genes, separately. The function of target genes was annotated by gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis. The expression profiles of nine selected miRNAs and their target genes verified that their expression patterns were opposite. This study has documented that analysis of miRNA under PEG and PEG + SNP conditions provides important insights into the improvement of drought resistance of alfalfa by exogenous NO at the molecular level. This has important scientific value and practical significance for the improvement of plant drought resistance by exogenous NO.

Combination Surgery of Silicone Tube Intubation and Conjunctival Resection in Patients with Epiphora.

PURPOSE: To compare the success rates of performing only silicone tube intubation versus carrying out both conjunctival resection and silicone tube intubation. METHODS: The subjects of this study involved 62 patients (96 eyes) between October 2015 and May 2017 who were diagnosed as having punctal stricture or nasolacrimal duct stenosis. Out of 96 eyes, 47 underwent only silicone tube intubation, and 49 underwent both silicone tube intubation and conjunctival resection. Three parameters were measured at 1, 3, and 6 months after the surgery: the area of the tear meniscus using RTVue-100 anterior segment optical coherence tomography, the height of the tear meniscus using a slit lamp microscope, and the subjective satisfaction of patients as a result of improved sympotms like epiphora. The surgery was considered successful when the patients' experienced the resolution of symptoms and reduction of the area and height of the tear meniscus. RESULTS: The area of the tear meniscus, height of the tear meniscus, and subjective satisfaction of patients was superior in the group that underwent both silicone tube intubation and conjunctival resection compared silicone tube intubation only. Based on these results, the success rate of the surgery was 68.9% in the group that underwent only silicone tube intubation and 78.7% in the group that underwent both silicone tube intubation and conjunctival resection. CONCLUSIONS: The resection of relaxed plica semilunares seems to increase the success rate of silicone tube intubation through the reduction of the area and height of the tear meniscus. Therefore, after determining the degree of conjunctivochalasis, if it was found to be severe, a combination with conjunctival resection was expected to increase the success rate of the surgery.

Effects of added polyacrylamide on changes in water states during the composting of kitchen waste.

The effects of adding polyacrylamide (PAM), to attempt to delay the loss of capillary water and achieve a better level of organic matter humification, in the composting of kitchen waste were evaluated. Four treatments, with initial moisture content of 60 % were used: 0.1 % PAM added before the start of composting (R1), 0.1 % PAM added when the thermophilic phase of composting became stable (at >50 °C) (R2), 0.1 % PAM added when the moisture content significantly decreased (R3), and no PAM added (R4). The introduction of PAM in R1 and R2 significantly increased the capillary force and delayed the loss of moisture content and capillary water. The introduction of PAM in R2 and R3 improved the composting process, in terms of the degradation of biochemical fractions and the humification degree. These results show that the optimal time for adding PAM was the initial stage of the thermophilic phase.