Anticancer and bone-enhanced nano-hydroxyapatite/gelatin/polylactic acid fibrous membrane with dual drug delivery and sequential release for osteosarcoma.

Surgical resection of osteosarcoma is always accompanied by residual metastasis of tumor cells and bone tissue defects. In this work, a novel kind of gelatin/polylactic acid (PLA) coaxial fiber membrane with a shell layer containing doxorubicin-loaded hydroxyapatite (DOX@nHAp) nanoparticles and a core layer containing Icariin (ICA) was developed for antitumor and bone enhancement at the defect site. Physical evaluation displayed that the composite membrane provided moderate hydrophilicity, enhanced tensile strength (Dry: 2-3 MPa, wet: 1-2 MPa) and elasticity (70-100 %), as well as increased specific surface area and pore volume (19.39 m2/g and 0.16 cm3/g). In SBF, DOX@nHAp in the fibers promoted biomineralization on the fiber surface. In in vitro evaluation, approximately 80 % of DOX had a short-term release during the first 8 days, followed by long-term release behavior of ICA for up to 40 days. CCK-8 results confirmed that the membrane could actively support MC3T3-E1 cells proliferation and was conductive to high alkaline phosphatase expression, while the viability of MG-63 cells was effectively inhibited to 50 %. Thus, the dual-loaded fibrous membrane with a coaxial structure and nHAp is a promising system for anticancer and defects reconstruction after osteosarcoma surgery.

Poly(ß-amino ester) Dual-Drug-Loaded Hydrogels with Antibacterial and Osteogenic Properties for Bone Repair.

In this study, we developed a poly(ß-amino ester) (PBAE) hydrogel for the double release of vancomycin (VAN) and total flavonoids of Rhizoma Drynariae (TFRD). VAN was covalently bonded to PBAE polymer chains and was released to enhance the antimicrobial effect first. TFRD chitosan (CS) microspheres were physically dispersed in the scaffold, TFRD was released from the microspheres, and osteogenesis was induced subsequently. The scaffold had good porosity (90.12 ± 3.27%), and the cumulative release rate of the two drugs in PBS (pH 7.4) solution exceeded 80%. In vitro antimicrobial assays demonstrated the antibacterial properties of the scaffold against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Besides these, cell viability assays indicated that the scaffold had good biocompatibility. Moreover, alkaline phosphatase and matrix mineralization were expressed more than in the control group. Overall, cell experiments confirmed that the scaffolds have enhanced osteogenic differentiation capabilities. In conclusion, the dual-drug-loaded scaffold with antibacterial and bone regeneration effects is promising in the field of bone repair.

Correction: Preparation and characterization of a novel drug-loaded Bi-layer scaffold for cartilage regeneration.

[This corrects the article DOI: 10.1039/D2RA00311B.].

Preparation and characterization of a novel drug-loaded Bi-layer scaffold for cartilage regeneration.

The incidence of articular cartilage defects is increasing year by year. In order to repair the cartilage tissue at the defect, scaffolds with nanofiber structure and biocompatibility have become a research hotspot. In this study, we designed and fabricated a bi-layer scaffold prepared from an upper layer of drug-dispersed gelatin methacrylate (GELMA) hydrogel and a lower layer of a drug-encapsulated coaxial fiber scaffold prepared from silk fiber (SF) and polylactic acid (PLA). These bi-layer scaffolds have porosity (91.26 ± 3.94%) sufficient to support material exchange and pore size suitable for cell culture and infiltration, as well as mechanical properties (2.65 ± 0.31 MPa) that meet the requirements of cartilage tissue engineering. The coaxial fiber structure exhibited excellent drug release properties, maintaining drug release for 14 days in PBS. In vitro experiments indicated that the scaffolds were not toxic to cells and were amenable to chondrocyte migration. Notably, the growth of cells in a bi-layer scaffold presented two states. In the hydrogel layer, cells grow through interconnected pores and take on a connective tissue-like shape. In the coaxial fiber layer, cells grow on the surface of the coaxial fiber mats and appeared tablet-like. This is similar to the structure of the functional partitions of natural cartilage tissue. Together, the bi-layer scaffold can play a positive role in cartilage regeneration, which could be a potential therapeutic choice to solve the current problems of clinical cartilage repair.

PDLLA/ß-TCP/HA/CHS/NGF Sustained-release Conduits for Peripheral Nerve Regeneration.

Using nerve guide conduits (NGCs) to promote the regeneration of PNI is a feasible alternative to autograft. Compared with NGCs made of single material, composite NGCs have a greater development prospect. Our previous research has confirmed that poly(D, L-lactic acid)/ß-tricalcium phosphate/hyaluronic acid/chitosan/nerve growth factor (PDLLA/ß-TCP/HA/CHS/NGF) NGCs have excellent physical and chemical properties, which can slowly release NGF and support cell adhesion and proliferation. In this study, PDLLA/ß-TCP/HA/CHS/NGF NGCs were prepared and used to bridge a 10 mm sciatic nerve defect in 200-250 g Sprague-Dawley (SD) rat to verify the performance of the NGCs in vivo. Substantial improvements in nerve regeneration were observed after using the PDLLA/ß-TCP/HA/CHS/NGF NGCs based on gross post-operation observation, triceps wet weight analysis and nerve histological assessment. In vivo studies illustrate that the PDLLA/ß-TCP/HA/CHS/NGF sustained-release NGCs can effectively promote peripheral nerve regeneration, and the effect is similar to that of autograft.

Development of a nanobody-based immunoassay for the sensitive detection of fibrinogen-like protein 1.

Immune checkpoint inhibition is an important strategy in cancer therapy. Blockade of CTLA-4 and PD-1/PD-L1 is well developed in clinical practice. In the last few years, LAG-3 has received much interest as an emerging novel target in immunotherapy. It was recently reported that FGL1 is a major ligand of LAG-3, which is normally secreted by the liver but is upregulated in several human cancers. FGL1 is a crucial biomarker and target for cancer immunotherapy. As the efficacy of immunotherapy is limited to specific types of patients, the subset of patients needs to be selected appropriately to receive precise treatment according to different biomarkers. To date, there is no test to accurately assess FGL1 expression levels. Nanobodies have some outstanding features, such as high stability, solubility and affinity for diagnostic and therapeutic applications. Here, we report the development and validation of a rapid, sensitive, and cost-effective nanobody-based immunoassay for the detection of FGL1 in human serum. In this study, human FGL1 recombinant protein was expressed and purified for the first time as an immunized antigen. Then, we constructed a nanobody phage display library and screened several nanobodies that bind FGL1 with high affinity. We selected two nanobodies targeting different epitopes of FGL1, one as a capture and the other conjugated with HRP as a probe. The double nanobody-based sandwich ELISA to detect the concentration of FGL1 showed a good response relationship in the range of 15.625-2000 ng/mL, and the recoveries from the spiked sample were in the range of 78% and 100%. This assay could be used as a potential approach for evaluating FGL1 expression for patient stratification and for predicting the therapeutic efficacy of targeting the LAG3/FGL1 axis.

Hyaluronic acid targeted and pH-responsive nanocarriers based on hollow mesoporous silica nanoparticles for chemo-photodynamic combination therapy.

In this work, a pH-responsive and tumor targeted multifunctional drug delivery system (RB-DOX@HMSNs-N = C-HA) was designed to realize chemo-photodynamic combination therapy. Hollow mesoporous silica nanoparticles (HMSNs) was served as the host material to encapsulate doxorubicin (DOX) and photosensitizer rose bengal (RB). Hyaluronic acid (HA) was modified on the surface of HMSNs via pH-sensitive Schiff base bonds as gatekeeper as well as targeted agent. Characterization results indicated the successful preparation of HMSNs-N = C-HA with appropriate diameter of 170 nm around and the nanocarriers displayed superior drug loading capacity (15.30 % for DOX and 12.78 % for RB). Notably, the results of in vitro drug release experiments confirmed that the system possessed good pH-sensitivity, which made it possible to release cargoes in slight acid tumor micro-environments. Significantly, the in vitro cell uptake and cytotoxicity assay results fully proved that RB-DOX@HMSNs-N = C-HA could precisely target murine mammary carcinoma (4T1) cells and effectively inhibit tumor cells viability with chemo-photodynamic synergistic therapy. Overall, our work (RB-DOX@HMSNs-N = C-HA) provides an efficient approach for the development of chemo-photodynamic combination therapy.

The electrostimulation and scar inhibition effect of chitosan/oxidized hydroxyethyl cellulose/reduced graphene oxide/asiaticoside liposome based hydrogel on peripheral nerve regeneration in vitro.

The application of hollow nerve conduits in the repair of peripheral nerve defects is effected by inferior recovery, and nerve extension is hampered by the scar tissue generated during the repair process. In this study, the filler in hollow nerve conduit, chitosan/oxidized hydroxyethyl cellulose (CS/OHEC) hydrogel loaded asiaticoside liposome and the conductive reduced graphene oxide (rGO) were developed and used to reform the microenvironment for peripheral nerve regeneration. The physiochemical properties of CS/OHEC/rGO/asiaticoside liposome hydrogel were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and compressive modulus, porosity, swelling ratio, degradation and conductivity. In addition, the asiaticoside release profiles in vitro were investigated. The hydrogel had a continuous porous network structure with pore size distribution in the range of 50-250 µm. The majority of the hydrogels had porosities above 70%, and a compressive modulus of 0.45 MPa. The weight loss rate of hydrogel reached 76.14 ± 4.45% within 8 weeks. The conductivity of the hydrogel was 5.27 ± 0.42 × 10-4 S/cm. The hydrogel was non-toxic and suitable for adhesion and proliferation of nerve cells in vitro. In addition, the application of electrical stimulation after the addition of rGO can promote the differentiation and proliferation of nerve cells, accelerating nerve regeneration. The asiaticoside released from the hydrogel had a significant inhibitory effect on the growth and collagen secretion of fibroblasts, eliminating scars for regenerative nerves, which can promote the function recovery of defected peripheral nerve. Together, these positive results indicate that the hydrogel would be a promising candidate for peripheral nerve regeneration.

Polydopamine coated hollow mesoporous silica nanoparticles as pH-sensitive nanocarriers for overcoming multidrug resistance.

A nanocarrier system of methoxypolyethylene glycol amine (mPEG-NH2) functionalized polydopamine (PDA) coated hollow mesoporous silica nanoparticles (HMSNs-PDA-PEG) was developed with pH-responsive, which combined doxorubicin hydrochloride (DOX) and quercetin (QUR) to reverse multidrug resistance (MDR) and improved anticancer effects on taxol (TAX) and DOX double resistant human colorectal cancer cell line HCT-8 (HCT-8/TAX cells). Well-dispersed nanoparticles (HMSNs-PDA-PEG) were prepared with a dimension of around 170 nm. The surface morphology and chemical properties of HMSNs-PDA-PEG were also successfully characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), thermal gravimetric analysis (TGA), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) method, Fourier transform infrared spectroscopy (FT-IR) and dynamic light scattering (DLS). Drug release experiments results indicated that DOX and QUR (QD) loaded nanoparticles (HMSNs-PDA-PEG@QD) had similar release kinetic profiles of each drug, which all exhibited highly sensitive to pH value due to the surface PDA coating. Additionally, the HCT-8 cells or HCT-8/TAX cells were employed to assess the cellular uptake and cytotoxicity of various drug-free or drug-loaded HMSNs samples. Meanwhile, a series of biological evaluations demonstrated that the HMSNs-PDA-PEG@QD exhibited remarkable ability to overcome MDR compared with free DOX and HMSNs-PDA-PEG@DOX. Taken together, these results revealed that HMSNs-PDA-PEG@QD was suitable as a prospective and efficient drug delivery nanosystem for overcoming multidrug resistance.

A dual-responsive, hyaluronic acid targeted drug delivery system based on hollow mesoporous silica nanoparticles for cancer therapy.

In this study, a novel drug delivery system (HMSNs-SS-HA) based on hollow mesoporous silica nanoparticles (HMSNs) was developed for delivering anticancer drugs (e.g., doxorubicin (DOX)) to targeted tumour cells by using disulfide bonds as redox-sensitive linkers and hyaluronic acid (HA) molecules as both capping and targeting agents. Well-dispersed HMSNs were synthesized with a dimension of around 100 nm. Detailed physical characterization further demonstrated that HMSNs-SS-HA has been successfully constructed. The in vitro drug release experiments displayed the enzyme and redox dual-responsive and sustained drug release properties of DOX loaded HMSNs-SS-HA. Additionally, a series of biological evaluations indicated that these DOX loaded HMSNs-SS-HA could accurately target murine mammary carcinoma (4T1) cells to induce cell apoptosis in vitro and suppress tumour growth in vivo. These results demonstrated that DOX loaded HMSNs-SS-HA was suitable as a potential and efficient drug delivery nanosystem for cancer therapy.

A conductive sodium alginate and carboxymethyl chitosan hydrogel doped with polypyrrole for peripheral nerve regeneration.

Polymer materials with electrically conductive properties have good applications in their respective fields because of their special properties. However, they usually exhibited poor mechanical properties and biocompatibility. In this work, we present a simple approach to prepare conductive sodium alginate (SA) and carboxymethyl chitosan (CMCS) polymer hydrogels (SA/CMCS/PPy) that can provide sufficient help for peripheral nerve regeneration. SA/CMCS hydrogel was cross-linked by calcium ions provided by the sustained release system consisting of d-glucono-δ-lactone (GDL) and superfine calcium carbonate (CaCO3), and the conductivity of the hydrogel was provided by doped with polypyrrole (PPy). Gelation time, swelling ratio, porosity and Young's modulus of the conductive SA/CMCS/PPy hydrogel were adjusted by polypyrrole content, and the conductivity of it was within 2.41 × 10-5 to 8.03 × 10-3 S cm-1. The advantages of conductive hydrogels in cell growth were verified by controlling electrical stimulation of cell experiments, and the hydrogels were also used as a filling material for the nerve conduit in animal experiments. The SA/CMCS/PPy conductive hydrogel showed good biocompatibility and repair features as a bioactive biomaterial, we expect this conductive hydrogel will have a good potential in the neural tissue engineering.

Dual targeting hyaluronic acid - RGD mesoporous silica coated gold nanorods for chemo-photothermal cancer therapy.

This work reports a multifunctional nanoplatform (GNRs@mSiO2-HA-RGD) by conjugating targeting ligand hyaluronic acid (HA) and RGD with mesoporous silica-coated gold nanorods (GNRs@mSiO2) for dual-targeted chemo-photothermal therapy. Doxorubicin hydrochloride (DOX) was used as the model drug to investigate the drug loading, in vitro drug release profiles and cell evaluation. The nanoplatform has demonstrated a good photothermal effect and excellent drug loading capacity of about 20.16%. It also had pH-enzyme sensitive and NIR-triggered drug release properties. Cellular uptake experiment results indicated that DOX-GNRs@mSiO2-HA-RGD can be dual-targeted to ovarian cancer cells via CD44 and integrin receptor mediated endocytosis pathway. Cytotoxicity experiments demonstrated that combined therapy exhibited a better therapy effect compared to that of single chemotherapy or photothermal therapy. Our study demonstrates that DOX-GNRs@mSiO2-HA-RGD may be a new promising dual-targeted delivery system for chemo-photothermal therapy.

Hyaluronic acid and Arg-Gly-Asp peptide modified Graphene oxide with dual receptor-targeting function for cancer therapy.

Graphene oxide (GO) modified with hyaluronic acid (HA) and Arg-gly-asp peptide (RGD) was designed as a dual-receptor targeting drug delivery system to enhance the specificity and efficiency of anticancer drug delivery. Firstly, GO-HA-RGD conjugate was characterized to reveal its structure and morphology. Whereafter, doxorubicin (Dox) as a model drug was loaded on GO-HA-RGD carrier, which displayed a high loading rate (72.9%, GO:Dox (w/w) = 1:1), pH-response and sustained drug release behavior. Cytotoxicity experiments showed that GO-HA-RGD possessed excellent biocompatibility towards SKOV-3 and HOSEpiC cells. Additionally, the GO-HA-RGD/Dox had a stronger cytotoxicity for SKOV-3 cells than either GO-HA/Dox (single receptor) or GO/Dox (no receptor). Moreover, celluar uptake studies illustrated that GO-HA-RGD conjugate could be effectively taken up by SKOV-3 cells via a synergic effect of CD44-HA and integrin-RGD mediated endocytosis. Hence, GO-HA-RGD nanocarrier is able to be a promising platform for targeted cancer therapeutic.

Synthesis and in vitro evaluation of a hyaluronic acid-quantum dots-melphalan conjugate.

Polymer-drug conjugates have played an important role in improving tumor cell targeting and the selectivity of anticancer drugs. In this study, quantum dots and melphalan were attached to the backbone of hyaluronic acid to synthesize a polymer-drug conjugate. The physicochemical properties of the conjugate were characterized by FT-IR, XRD, (1)H NMR, UV-Vis spectra and DLS. The in vitro drug release profiles and cell evaluation were investigated. The results showed that the conjugate was synthesized and self-assembled into nanoparticles with a diameter of 115 ± 2.3 nm. The conjugate had a pH-sensitive drug controlled release property. It was an ideal receptor-mediated delivery system and can be internalized into the human breast cancer cell. It had a better inhibition effect on human breast cancer cell and a poorer inhibition effect on normal breast cell than melphalan. These results supported that the conjugate would be a promising candidate for cancer therapy.

[Identification on Xanthium sibiricum from different habitats by FTIR fingerprint patterns].

OBJECTIVE: To establish an FTIR method to identify Xanthium sibiricum from different habitats. METHODS: FTIR spectra of Xanthium sibiricum from different habitats were analyzed,and the similarity of different fingerprint spectra and the chemical pattern recognition were calculated and analyzed according to the wave numbers of peaks. RESULTS: Different FTIR spectra of 10 different habitats of Xanthium sibiricum were obtained,and the similarities were all above 0. 96. CONCLUSION: This method can be used for identification on Xanthium sibiricum from different habitats. The results of similarity calculation and chemical pattern recognition further prove the feasibility of this method.

A novel melphalan polymeric prodrug: preparation and property study.

The clinical application of melphalan (Me), an anticancer drug for the treatment of hematologic malignancies, has been limited due to its poor water solubility, rapid elimination and lack of target specificity. To solve these problems, O,N-carboxymethyl chitosan-peptide-melphalan conjugates were synthesized and characterized. All polymeric prodrugs showed satisfactory water solubility. It was found that the molecular weight of O,N-carboxymethyl chitosan (O,N-CMCS) and the peptide spacer played a crucial role in controlling the drug content, diameter and drug release properties of O,N-carboxymethyl chitosan-peptide-melphalan conjugates. The studies of in vitro drug release and cell cytotoxicity by MTT assay revealed that, employing the polymeric conjugation strategy and using the peptides glycylglycine (Gly-Gly) as a spacer, the conjugates have good cathepsin X-sensitivity and lower toxicity and the drug release behavior improved remarkably. In conclusion, O,N-carboxymethyl chitosan-peptide-melphalan conjugates could be promising prodrugs for anticancer application.

Conductive PPY/PDLLA conduit for peripheral nerve regeneration.

The significant drawbacks and lack of success associated with current methods to treat critically sized nerve defects have led to increased interest in neural tissue engineering. Conducting polymers show great promise due to their electrical properties, and in the case of polypyrrole (PPY), its cell compatibility as well. Thus, the goal of this study is to synthesize a conducting composite nerve conduit with PPY and poly(d, l-lactic acid) (PDLLA), assess its ability to support the differentiation of rat pheochromocytoma 12 (PC12) cells in vitro, and determine its ability to promote nerve regeneration in vivo. Different amounts of PPY (5%, 10%, and 15%) are used to synthesize the conduits resulting in different conductivities (5.65, 10.40, and 15.56 ms/cm, respectively). When PC12 cells are seeded on these conduits and stimulated with 100 mV for 2 h, there is a marked increase in both the percentage of neurite-bearing cells and the median neurite length as the content of PPY increased. More importantly, when the PPY/PDLLA nerve conduit was used to repair a rat sciatic nerve defect it performed similarly to the gold standard autologous graft. These promising results illustrate the potential that this PPY/PDLLA conducting composite conduit has for neural tissue engineering.

Preparation, characterization, and in vitro efficacy of O-carboxymethyl chitosan conjugate of melphalan.

A series of melphalan-O-carboxymethyl chitosan (Mel-OCM-chitosan) conjugates with different spacers were prepared and structurally characterized. All conjugates showed satisfactory water-solubility (160-217 times of Mel solubility). In vitro drug release behaviors by both chemical and enzymatic hydrolysis were investigated. The prodrugs released Mel rapidly within papain and lysosomal enzymes of about 40-75%, while released only about 4-5% in buffer and plasma, which suggested that the conjugates have good plasma stability and the hydrolysis in both papain and lysosomes occurs mostly via enzymolysis. It was found that the spacers have important effect on the drug content, water solubility, drug release properties and cytotoxicity of Mel-OCM-chitosan conjugates. Cytotoxicity studies by MTT assay demonstrated that these conjugates had 52-70% of cytotoxicity against RPMI8226 cells in vitro as compared with free Mel, indicating the conjugates did not lose anti-cancer activity of Mel. Overall these studies indicated Mel-OCM-chitosan conjugates as potential prodrugs for cancer treatment.

[Identification on Solanum lyratum by FT-IR fingerprint patterns and similarity degree of different fingerprint patterns].

OBJECTIVE: To provide a FT-IR new method to identify different habitats of Solanum lyratum. METHODS: Analyzed FT-IR patterns of Solanum lyratum from different habitats, and the similarity of different fingerprint patterns was calculated and analyzed according to the wave numbers of peaks searched. RESULTS: Obtained the different FT-IR pattern of 5 different habitats of Solanum lyraturn. CONCLUSION: This method can be used for identifications on different habitats of Solanum lyratum. The results of similarity calculation further prove the feasibility of this method.

Preparation, characterization and in vitro release study of a glutathione-dependent polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan.

In this work, an amphiphilic polymeric prodrug Cis-3-(9H-purin-6-ylthio)-acrylic acid-graft-carboxymethyl chitosan (PTA-g-CMCS) was designed and synthesized. In aqueous solution, this grafted polymer can self-assemble into spherical micelles with a size ranging from 104 to 285 nm and zeta potential ranging from -12.3 to -20.1 mV. For the release study, less than 24% of 6-Mercaptopurine (6-MP) was released from PTA-g-CMCS1 in the media containing 2 and 100 µM glutathione (GSH), whereas 37%, 54% and 75% of 6-MP was released from the media with GSH of 1, 2 and 10mM, respectively. Besides, pH and drug content of the polymeric prodrug only presented slight influence on the 6-MP release. MTT assay demonstrated that this system had higher inhibition ratio on HL-60 cells (human promyelocytic leukemia cells) in the presence of GSH and lower cytotoxicity on mouse fibroblast cell line (L929). Therefore, this nano-sized system is glutathione-dependent, and it can be employed as a potential carrier for the controlled release of 6-MP.