Haixing Xu

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.

PDLLA/chondroitin sulfate/chitosan/NGF conduits for peripheral nerve regeneration.

Biodegradable PDLLA/Chondroitin sulfate/Chitosan(PDLLA/CS/CHS) nerve conduits with potentially good biocompatibility and good mechanical property feasible for surgical manipulation have been developed in our previous work. The purpose of this study was to investigate their possible application in repairing damaged nerves and the effect of nerve growth factor (NGF). The PDLLA/CS/CHS/NGF nerve conduits were prepared by immobilizing NGF onto the PDLLA/CS/CHS nerve conduits with carbodiimide. Adult Sprague-Dawley (SD) rats weighing 200-250 g were used as the animal model. The conduits were employed to bridge the 10 mm defects in the sciatic nerve of the SD rats. Nerve conduction velocities (NCVs) were clearly detected in both nerve conduits after 3 months of implantation, indicating a rapid functional recovery for the disrupted nerves. The results of histological sections showed that the internal sides of the conduits were compact enough to prevent the connective tissues from ingrowth. Combined with the strong mechanical properties, good nerve regeneration ability and non-toxicity of its degradation products, PDLLA/CS/CHS nerve conduits would be expected to be useful materials to repair nerve damage and NGF can effectively promote the regeneration of peripheral nerve defect.

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.

Preparation and characterization of a dual-receptor mesoporous silica nanoparticle–hyaluronic acid–RGD peptide targeting drug delivery system

A dual-receptor targeting delivery system based on mesoporous silica nanoparticles modified with hyaluronic acid and RGD peptide (MSNs/NH2–HA–RGD) was developed in the present study, and characterized by TEM, SAXRD, nitrogen adsorption–desorption analysis, DLS, FT-IR, 13C NMR and UV-vis. The results showed that MSNs/NH2–HA–RGD had an ideal monodispersibility, uniform particle size (172.5 ± 10 nm) and well-defined mesoporous structure. Moreover, cellular uptake results showed that MSNs/NH2–HA–RGD had an ideal dual-receptor mediated endocytosis pathway and could be significantly internalized into ovarian cancer cells. Chlorambucil (CHL), an anticancer drug, was chosen as a model drug to investigate drug loading, in vitro drug release behaviors and cytotoxicity. The results showed that CHL-loaded MSNs/NH2–HA–RGD exhibited a high drug loading capacity of about 10.1% and pH-sensitive drug controlled release behaviors. The cytotoxicity test showed that CHL-loaded MSNs/NH2–HA–RGD had a stronger cytotoxicity for ovarian cancer cells than one receptor or no receptor modified MSNs. It is expected that MSNs/NH2–HA–RGD may be a prospective candidate for targeted delivery of anticancer drugs to cancer foci.

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.

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.

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.

Degradation properties of the electrostatic assembly PDLLA/CS/CHS nerve conduit.

A poly(d,l-lactic acid)/chondroitin sulfate/chitosan (PDLLA/CS/CHS) nerve conduit for repairing nerve defects was prepared by electrostatic assembly and the thermally induced phase separation technique. The hydrophilic characteristics of the PDLLA/CS/CHS assembly nerve conduits were improved markedly. The degradation behavior of the nerve conduit with various assembly layers was evaluated by a pH change, weight loss rate and molecular weight change. The pH of the solution of the nerve conduit could be effectively adjusted by varying the layer numbers and overcoming the acidity-caused auto-acceleration of PDLLA; the nerve conduit can retain its integrity in a phosphate buffer solution after being degraded for 3 months. After such a conduit was implanted in the rat for 3 months, obvious degradation occurred, but the regenerated nerve was integrated and it grew successfully from the proximal to distal nerve stump. All these results implied that the degradation rate of the prepared conduit can adapt to the regeneration of the peripheral nerve, which might be a new derivative of PDLLA-based biodegradable materials for repairing nerve injuries without acidity-caused irritations and acidity-induced auto-accelerating degradation behavior as shown by PDLLA.

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 methotrexate conjugated O, N-carboxymethyl chitosan via peptidyl spacers.

The use of methotrexate (MTX), an anticancer drug for the treatment of hematologic malignancies, has been limited in the clinical application due to its poor water solubility, high clearance rate, and lack of target specificity. To solve these problems, O,N-carboxymethyl chitosan–dipeptide-MTX conjugates have been synthesized and characterized by fourier transform infrared radiation spectroscopy and proton nuclear magnetic resonance (1H NMR). All polymeric conjugates showed satisfactory water solubility. The results of the study revealed that drug release and toxicity were affected by employing polymeric conjugation strategy and dipeptide spacers [glycylglycine (Gly–Gly), glycyl–l-phenylalanine (Gly–Phe), glycyl–l-tyrosine (Gly–Tyr)]. It has been found that drugs could be effectively loaded and released when polymeric prodrugs were combined with a dipeptide spacer. In conclusion, O,N-CMCS–dipeptide-MTX polymeric prodrugs could potentially be used as responsive drug delivery systems.