Controlled release of cisplatin and cancer cell apoptosis with cisplatin encapsulated poly(lactic-co-glycolic acid) nanoparticles

The goal of the present study is to utilize cis-diamminedichloroplatinum (cisplatin) loaded polymer nanoparticles (NPs) to give a controlled, extended, and local drug therapy for the treatment of cancer. We have used biodegradable and biocompatible poly(lactic-co-glycolic acid) (PLGA) to prepare the NPs by adjusting the double emulsion technique using poly(vinylalcohol) as a surface active agent. The PLGA NPs were characterized for particle size and shape, controlled release of cisplatin, and degradation. Cisplatin solubility in deionized water was increased up to 4 mg/mL by simply changing the solution parameters. Cisplatin encapsulated NPs were incubated in phosphate buffered saline (PBS) at 37?C to study the release kinetics of cisplatin. Cisplatin was released in a sustained manner with less than 20% release during a 3-day period followed by 50% release during a 21-day period. A degradation study of PLGA NPs demonstrated the loss of spherical shape during a 21-day period. We also examined the cisplatin sensitive A2780 cell apoptosis when cells were incubated with cisplatin encapsulated PLGA NPs. A large number of cell apoptosis occurred as a result of cisplatin release from the PLGA NPs. These results suggest that cisplatin encapsulated PLGA NPs can be used to treat the cancer cells by injecting them into a localized site minimizing the side effects.

Viscoelasticity of repaired sciatic nerve by poly(lactic-co-glycolic acid) tubes

Medical-grade synthetic poly（lactic-co-glycolic acid） polymer can be used as a biomaterial for nerve repair because of its good biocompatibility, biodegradability and adjustable degradation rate. The stress relaxation and creep properties of peripheral nerve can be greatly improved by repair with poly（lactic-co-glycolic acid） tubes. ＂Fen sciatic nerve specimens were harvested from fresh corpses within 24 hours of death, and were prepared into sciatic nerve injury models by creating a 10 mm defect in each specimen. Defects were repaired by anastomosis with nerve autografts and poly（lactic-co-glycolic acid） tubes. Stress relaxation and creep testing showed that at 7 200 seconds the sciatic nerve anastomosed by poly（lactic-co-glycolic acid） tubes exhibited a greater decrease in stress and increase in strain than those anastomosed by nerve autografts. These findings suggest that poly（lactic-co-glycolic acid） exhibits good viscoelasticity to meet the biomechanical require- ments for a biomaterial used to repair sciatic nerve injury.

A novel artificial nerve graft for repairing longdistance sciatic nerve defects:a self-assembling peptide nanofiber scaffold-containing poly (lactic-co-glycolic acid) conduit

In this study, we developed a novel artificial nerve graft termed self-assembling peptide nanofiber scaffold （SAPNS）-containing poly（lactic-co-glycolic acid） （PLGA） conduit （SPC） and used it to bridge a 10-mm-long sciatic nerve defect in the rat. Retrograde tracing, behavioral testing and histomorphometric analyses showed that compared with the empty PLGA conduit implantation group, the SPC implantation group had a larger number of growing and extending axons, a markedly increased diameter of regenerated axons and a greater thickness of the myelin sheath in the conduit. Furthermore, there was an increase in the size of the neuromuscular junction and myofiber diameter in the target muscle. These findings suggest that the novel artificial SPC nerve graft can promote axonal regeneration and remyelination in the transected peripheral nerve and can be used for repairing peripheral nerve injury.

Dorsal root ganglion-derived Schwann cells combined with poly(lactic-co-glycolic acid)/chitosan conduits for the repair of sciatic nerve defects in rats

Schwann cells, nerve regeneration promoters in peripheral nerve tissue engineering, can be used to repair both the peripheral and central nervous systems. However, isolation and puriifcation of Schwann cells are complicated by contamination with ifbroblasts. Current reported measures are mainly limited by either high cost or complicated procedures with low cell yields or purity. In this study, we collected dorsal root ganglia from neonatal rats from which we obtained highly puriifed Schwann cells using serum-free melanocyte culture medium. The purity of Schwann cells （〉95%） using our method was higher than that using standard medium containing fetal bovine serum. The obtained Schwann cells were implanted into poly（lactic-co-glycolic acid）/chi-tosan conduits to repair 10-mm sciatic nerve defects in rats. Results showed that axonal diameter and area were signiifcantly increased and motor functions were obviously improved in the rat sciatic nerve tissue. Experimental ifndings suggest that serum-free melanocyte culture medium is conducive to purify Schwann cells and poly（lactic-co-glycolic acid）/chitosan nerve conduits combined with Schwann cells contribute to restore sciatic nerve defects.

Poly(lactic-co-glycolic acid) conduit for repair of injured sciatic nerve A mechanical analysis

Tensile stress and tensile strain directly affect the quality of nerve regeneration after bridging nerve defects by poly（lactic-co-glycolic acid） conduit transplantation and autogenous nerve grafting for sciatic nerve injury. This study collected the sciatic nerve from the gluteus maximus muscle from fresh human cadaver, and established 10-mm-long sciatic nerve injury models by removing the ischium, following which poly（lactic-co-glycolic acid） conduits or autogenous nerve grafts were transplanted. Scanning electron microscopy revealed that the axon and myelin sheath were torn, and the vessels of basilar membrane were obstructed in the poly（lactic-co-glycolic acid） conduit-repaired sciatic nerve following tensile testing. There were no significant differences in tensile tests with autogenous nerve graft-repaired sciatic nerve. Following poly（lactic-co-glycolic acid） conduit transplantation for sciatic nerve repair, tensile test results suggest that maximum tensile load, maximum stress, elastic limit load and elastic limit stress increased compared with autogenous nerve grafts, but elastic limit strain and maximum strain decreased. Moreover, the tendencies of stress-strain curves of sciatic nerves were similar after transplantation of poly（lactic-co-glycolic acid） conduits or autogenous nerve grafts. Results showed that after transplantation in vitro for sciatic nerve injury, poly（lactic-co-glycolic acid） conduits exhibited good intensity, elasticity and plasticity, indicating that poly（lactic-co-glycolic acid） conduits are suitable for sciatic nerve injury repair.

A Study of Surface Modification of Poly(lactic-co-glycolic) Acid Using Argon Ion Irradiation

The effect of Argon ion irradiation to the surface properties of poly(lactic-co-glycolic) acid (PLGA) was studied. A beam of 170 keV Argon ions was implanted at different fluencies (1 × 1012, 1 × 1013, 1 × 1014, and 1 × 1015 ions/cm2). X-ray photoelectron spectroscopy (XPS) was used to analyze the evolution of the bonding microstructure of PLGA due to irradiation. Surface morphology was monitored using atomic force microscopy (AFM). AFM analysis shows that film roughness increased to maximum at the dose of 1 × 1014 ions/cm2 where the formations of hillocks were also detected. Hydrophilicity of PLGA is important for their applications in biomedicine such as bioscaffolds. Hydrophilicity was monitored using water contact angle measurements for both unmodified and ion-modified PLGA. It was observed that hydrophilicity of PLGA changes with the ion irradiation. This demonstrates that ion irradiation can be an alternative approach to control hydrophilicity of PLGA. PLGA scaffolds modified with ion irradiation could therefore be more suitable for the biomedical applications.

Evaluation of in vitro and in vivo immunostimulatory activities of poly(lactic-co-glycolic acid) nanoparticles loaded with soluble and autoclaved Leishmania infantum antigens: A novel vaccine candidate against visceral leishmaniasis

Objective: To prepare and characterize poly lactic-co-glycolic acid(PLGA) nanoparticles loaded with soluble leishmanial antigen or autoclaved leishmanial antigen and explore in vitro and in vivo immunogenicity of antigen encapsulated nanoparticles. Methods: Water/oil/water double emulsion technique was employed to synthesize PLGA nanoparticles, and scanning electron microscopy, Fourier transform infrared spectroscopy and Zeta-potential measurements were used to identify the characteristics of nanoparticles. Cytotoxicity of synthetized nanoparticles on J774 macrophage were investigated by MTT assays. To determine the in vitro immunostimulatory efficacies of nanoparticles, griess reaction and ELISA was used to measure the amounts of NO and cytokines. During the in vivo analysis, Balb/c mice were immunized with vaccine formulations, and protective properties of nanoparticles were measured by Leishman Donovan unit in the liver following the infection. Cytokine levels in spleens of mice were determined by ELISA. Results: MTT assay showed that neither soluble leishmanial antigen nor autoclaved leishmanial antigen encapsulated nanoparticles showed cytotoxicity against J774 macrophage cells. Contrary to free antigens, both autoclaved leishmanial antigen-nanoparticle and soluble leishmanial antigen-nanoparticle formulations led to a 10 and 16-fold increase in NO amounts by macrophages, respectively. Leishman Donovan unit calculations revealed that soluble leishmanial antigen-nanoparticles and autoclaved leishmanial antigen-nanoparticles yielded 52% and 64% protection against visceral leishmaniasis in mouse models. Besides, in vitro and in vivo tests demonstrated that by increasing IFN-γ and IL-12 levels and inhibiting IL-4 and IL-10 secretions, autoclaved leishmanial antigen-nanoparticles and soluble leishmanial antigennanoparticles triggered Th1 immune response. Conclusions: Both autoclaved leishmanial antigen-nanoparticles and soluble leishmanial antigen-nanoparticles formulations provide exceptional in vitro and in vivo immunostimulatory activities. Hence, PLGA-based antigen delivery systems are recommended as potential vaccine candidates against visceral leishmaniasis.

Effects of poly lactic-co-glycolic acid-Nogo A antibody delayed-release microspheres on regeneration of injured spinal cord in rats

BACKGROUND： Nogo A antigen is the major inhibiting factor blocking regeneration of the injured spinal cord. Neutralizing Nogo A antigens using Nogo A antibodies may help promote neurite regeneration and nervous function recovery. For successful regeneration, sustained release of the antibody from a biodegradable material loaded with Nogo A antibodies to the injury site is required. OBJECTIVE： To compare the therapeutic effects of poly lactic-co-glycolic acid （PLGA）-Nogo A antibody delayed-release microspheres and Nogo A antibody alone on spinal regeneration in Sprague-Dawley rats with complete transverse injury to the spinal cord. DESIGN, TIME AND SETTING： A randomized, controlled animal trial was performed at the Pharmacological Laboratory of West China Center of Medical Sciences, Sichuan University, between October 2007 and January 2008. MATERIALS： Goat anti-rat Nogo A monoclonal antibody was purchased from Santa, American; goat anti-rat neurofilament 200 monoclonal antibody was from Zhongshan Goldenbridge, Beijing, China; PLGA-Nogo A antibody delayed-release microspheres were provided by the College of Pharmacy, Sichuan University. METHODS： A total of 36 adult female Sprague Dawley rats were used to establish models of completely transected spinal cord injury, at T10. Animals were randomly divided into three groups （n=12）： model, Nogo A antibody alone, and Nogo A antibody delayed-release microsphere groups. After transverse injury of the spinal cord, 50 μ L normal saline solution, 50 μL normal saline solution containing 50μL g Nogo A antibody, and 50 μL normal saline solution containing 50 μg Nogo A antibody microspheres were administered to the respective groups at the injury site. MAIN OUTCOME MEASURES： The expression of Nogo A and neurofilament 200 in injured spinal cord was tested immunohistochemically, and motor function of rats was assessed by Basso-Beattie-Bresnahan （BBB） locomotor rating scale. RESULTS： Four weeks after injury, expression of Nogo A in microsphere group was significantly less than model and Nogo A antibody alone groups （P 〈 0.05）; while there was no significant difference between model and Nogo A antibody alone groups （P 〉 0.05）. Ten weeks after injury, microsphere group showed a significantly greater expression of neurofilament 200 than model and Nogo A antibody alone groups （P 〈 0.05）; while no significant difference was found between model and Nogo A antibody alone groups （P 〉 0.05）. At postoperative weeks 5 and 6, the score of BBB locomotor rating scale in microsphere group was significantly greater than the model group （P 〈 0.05）, and at postoperative weeks 7 10, the score was much greater than model and Nogo A antibody alone groups （P 〈 0.05）. CONCLUSION： Nogo A antibody delayed-release microspheres decreased Nogo A expression, increased neurofilament 200 expression in the injured spinal cord of rats, and promoted recovery of motor function through sustained drug release over a long-term period.

A porous poly(lactic-co-glycolic acid) scaffold induces innervation in a rabbit model of disc degeneration following annular injury

BACKGROUND： A degradable poly（lactic-co-glycolic acid） （PLGA） scaffold has been used to construct a degradable porous scaffold. This template can simulate the in vivo microenvironment and promote tissue formation. OBJECTIVE： To observe the histopathological changes during degeneration and regeneration of the intervertebral disc, and to analyze the effects of a PLGA scaffold on nerve fiber ingrowth into the lesion in vivo. DESIGN, TIME AND SETTING： A randomized, controlled animal experiment was performed at the Orthopaedic Laboratory, Clinic Medical Research Institution, Sir Run Run Shaw Hospital, Zhejiang University, from December 2007 to July 2008. MATERIALS： PLGA （China Textile Academy）; growth-associated protein-43 （Life-span, USA）; and protein gene product 9.5 antibody （AbD, United Kingdom） were used in this study. METHODS： Three consecutive segments of the intervertebral disc of thirty-two healthy adult male New Zealand rabbits were exposed, comprising L3-4, L4-5 and L5-6. Experimental intervertebral disc （L4-5 and L5-6） models were established by two different methods. In the test （trephine ＋ scaffold） group, a 5-mm deep hole was drilled into the annulus fibrosus using a 3-mm diameter trephine, and the PLGA scaffold was implanted into the hole. In the acupuncture group, the remaining experimental intervertebral disc annulus fibrosus was damaged using a 16G needle at a depth of 5 mm. The L3-4 disc served as a control. MAIN OUTCOME MEASURES： Intervertebral disc degeneration was assessed using radiography, magnetic resonance imaging, and histological examination at various time points post-surgery. Nerve fiber ingrowth into the degenerated intervertebral disc was observed using immunohistochemical staining for growth-associated protein-43 and protein gene product 9.5. RESULTS： Compared with the normal controls, the heights of the damaged intervertebral discs were decreased, and T2 signal intensity was decreased in the test and acupuncture groups 2 weeks post-surgery. Intervertebral disc degeneration was faster in the test group than in the acupuncture group. PLGA was coated with newly formed tissue, gradually degraded, and absorbed, and could induce tissue ingrowth deep into the annulus fibrosus. Results of immunohistochemical staining showed that nerve fibers were distributed in newly formed tissue in the test group, and in the superficial layer or surrounding scar tissue in the acupuncture group. CONCLUSION： A porous PLGA scaffold provides an important biological channel to induce nerve fiber ingrowth deep into the degenerated intervertebral disc.

MMAE-loaded PLGA nanomedicine with improved biosafety to achieve efficient antitumor treatment

Monomethyl auristatin E(MMAE)is a derivative of the marine peptide Dolastatin 10,which has therapeutic effects against various cancers according to its antimitotic activity in multiple clinical trials.The antibody drug conjugate(ADC)of MMAE is currently used in clinical practice.However,the safety issues of MMAE-based ADC,such as high drug toxicity and poor bioavailability,still exist when using it for anticancer therapy.A sustained release of drug delivery approach should be used to reduce toxicity and achieve sufficient anticancer effects.Herein,PLGA-b-PEG 2000 with excellent biocompatibility and slow degradation ability was adopted to construct MMAE-loaded nanoparticles for safe and effective chemotherapy.The sustained release effect and the immunogenic cell death(ICD)effect of PLGA-MMAE nanoparticles were assessed by in vitro experiments.The PLGA-MMAE nanoparticles effectively accumulated in the tumor through the enhanced permeability and retention(EPR)effect,inducing cell apoptosis and causing a certain degree of immune response.The sustained drug release of PLGA-MMAE improved the bioavailability and effectively reduced the toxicity and development of the tumor compared to the effect of free MMAE or ADC.Overall,this study provides a safe and effective chemotherapeutic approach,as well as a simple and effective synthetic process for MMAE-based nanoparticles,improving their therapeutic efficacy and safety.

Synthesis,Characterization and Application of Poly(lactic-co-glycolic acid)with a Mass Ratio of Lactic to Glycolic Segments of 52/48

Poly(lactic-co-glycolic acid)(PLGA)is one of the most representative degradable copolymers and promising drug carriers.In the current paper,the PLGAs with a lactic acid/glycolic acid(LA/GA)molar ratio of 52/48 and various molecular weights were prepared by a melting method.The molecular weight,molecular weight distribution,and thermal stability were determined by 1H NMR and thermogravimetric analysis methods.The results demonstrated that PLGAs with the fixed LA/GA molar ratio(52/48),different molecular weights,and narrow molecular weight distribution could be obtained by solely altering the reaction time.The PLGA films were prepared,and their properties including micro-structure,mechanical property,in-vitro cytotoxicity,and biodegradability were characterized.In combination with the homogeneous microstructures and mechanical properties,the drug-loading and releasing properties of PLGA3.2 films were investigated.The results show that PLGA3.2 film with an LA/GA molar ratio of 52/48 is a promising curcumin carrier.

Poly(lactic-co-glycolic acid)-based composite bone-substitute materials

Research and development of the ideal artificial bone-substitute materials to replace autologous and allogeneic bones for repairing bone defects is still a challenge in clinical orthopedics.Recently,poly(lactic-co-glycolic acid)(PLGA)-based artificial bone-substitute materials are attracting increasing attention as the benefit of their suitable biocompatibility,degradability,mechanical properties,and capabilities to promote bone regeneration.In this article,we comprehensively review the artificial bone-substitute materials made from PLGA or the composites of PLGA and other organic and inorganic substances,elaborate on their applications for bone regeneration with or without bioactive factors,and prospect the challenges and opportunities in clinical bone regeneration.

Inhalable oridonin-loaded poly(lactic-co-glycolic)acid large porous microparticles for in situ treatment of primary non-small cell lung cancer

Non-small cell lung cancer(NSCLC) accounts for about 85% of all lung cancers. Traditional chemotherapy for this disease leads to serious side effects. Here we prepared an inhalable oridonin-loaded poly(lactic-co-glycolic)acid(PLGA) large porous microparticle(LPMP) for in situ treatment of NSCLC with the emulsion/solvent evaporation/freeze-drying method. The LPMPs were smooth spheres with many internal pores. Despite a geometric diameter of 10 mm, the aerodynamic diameter of the spheres was only 2.72 mm, leading to highly efficient lung deposition. In vitro studies showed that most of oridonin was released after 1 h, whereas the alveolar macrophage uptake of LPMPs occurred after 8 h, so that most of oridonin would enter the surroundings without undergoing phagocytosis. Rat primary NSCLC models were built and administered with saline, oridonin powder, gemcitabine, and oridonin-loaded LPMPs via airway, respectively. The LPMPs showed strong anticancer effects. Oridonin showed strong angiogenesis inhibition and apoptosis. Relevant mechanisms are thought to include oridonin-induced mitochondrial dysfunction accompanied by low mitochondrial membrane potentials, downregulation of BCL-2 expressions, upregulation of expressions of BAX, caspase-3 and caspase-9. The oridonin-loaded PLGALPMPs showed high anti-NSCLC effects after pulmonary delivery. In conclusion, LPMPs are promising dry powder inhalations for in situ treatment of lung cancer.

Preparation of poly(lactide-co-glycolide)microspheres and evaluation of pharmacokinetics and tissue distribution of BDMC-PLGA-MS in rats

The aim of the present study was to develop a novel long-acting Poly(lactic-co-glycolic acid)(PLGA)-based microspheres formulation of Bisdemethoxycurcum(BDMC) by emulsionsolvent evaporation method. Meanwhile, the effects of the volume ratio of the dispersed phase and continuous phase, the concentration of PLGA and PVA, the theoretical drug loading and stirring speed were investigated. The mean diameter of the microspheres was 8.5 μm and the size distribution was narrow. The encapsulation efficiency(EE) and drug loading efficiency(DLE) of BDME loaded PLGA microspheres(BDMC-PLGA-MS) was 94.18% and 8.14%,respectively. In an in vitro study of drug release, it can be concluded that the BDMC-PLGAMS exhibited sustained and long-term release properties for 96 h. Stability studies suggested that the microspheres we prepared had a very good stability. Furthermore, the results of an in vivo study indicated that the BDMC-PLGA-MS had sustained release effect and was mainly distributed in the lung tissue, and less distribution in other tissues, which indicated that microspheres could be an effective parenteral carrier for the delivery of BDMC in lung cancer treatment.

Mechanical Properties and Degradability of Electrospun PCL/PLGA Blended Scaffolds as Vascular Grafts

In our study, the mechanical properties and degradability of vascular grafts made from poly(ε-caprolactone)(PCL) and poly(lactic-co-glycolic acid)(PLGA) at different ratios were investigated. The results showed that the electrospun PCL/PLGA grafts possess good mechanical properties and biodegradability. The tensile and burst strength of the scaffolds met the demands of vascular grafts. In vitro degradation tests indicated that the degradation rate of the materials increased with the percentage of PLGA, and in vivo tests showed that increasing the amount of PLGA is an effective way to promote cell infiltration. Particularly, the electrospun PCL/PLGA blended scaffold with 10% PLGA exhibited a balance of mechanical and degradation properties, making it a promising tissue engineering material for vascular grafts.

Mathematical modeling of drug release from biodegradable polymeric microneedles

Transdermal drug delivery systems have overcome many limitations of other drug administration routes,such as injection pain and first-pass metabolism following oral route,although transdermal drug delivery systems are limited to drugs with low molecular weight.Hence,new emerging technology allowing high molecular weight drug delivery across the skin—known as‘microneedles’—has been developed,which creates microchannels that facilitate drug delivery.In this report,drug-loaded degradable conic microneedles are modeled to characterize the degradation rate and drug release profile.Since a lot of data are available for polylactic acid-co-glycolic acid(PLGA)degradation in the literature,PLGA of various molecular weights-as a biodegradable polymer in the polyester family-is used for modeling and verification of the drug delivery in themicroneedles.The main reaction occurring during polyester degradation is hydrolysis of steric bonds,leading to molecular weight reduction.The acid produced in the degradation has a catalytic effect on the reaction.Changes in water,acid and steric bond concentrations over time and for different radii of microneedles are investigated.To solve the partial and ordinary differential equations simultaneously,finite difference and Runge–Kutta methods are employed,respectively,with the aid of MATLAB.Correlation of the polymer degradation rate with its molecular weight and molecular weight changes versus time are illustrated.Also,drug diffusivity is related to matrix molecular weight.The molecular weight reduction and accumulative drug release within the system are predicted.In order to validate and assess the proposed model,data series of the hydrolytic degradation of aspirin(180.16 Da)-and albumin(66,000 Da)-loaded PLGA(1:1 molar ratio)are used for comparison.The proposed model is in good agreement with experimental data from the literature.Considering diffusion as themain phenomena and autocatalytic effects in the reaction,the drug release profile is predicted.Based on our results for a microneedle containing drug,we are able to estimate drug release rates before fabrication.

In vitro bioactivity and corrosion of PLGA/hardystonite composite-coated magnesium-based nanocomposite for implant applications

A type of polymer/ceramic coating was introduced on a magnesium-based nanocomposite, and the nanocomposite was evaluated for implant applications.The microstructure, corrosion, and bioactivity of the coated and uncoated samples were assessed.Mechanical alloying followed by sintering was applied to fabricate the Mg–3Zn–0.5Ag–15NiTi nanocomposite substrate.Moreover, different contents of poly(lactic-co-glycolic acid)(PLGA) coatings were studied, and 10 wt% of PLGA content was selected.The scanning electron microscopy(SEM) images of the bulk nanocomposite showed an acceptable homogenous dispersion of the Ni Ti nanoparticles(NPs) in the Mg-based matrix.In the in vitro bioactivity evaluation, following the immersion of the uncoated and coated samples in a simulated body fluid(SBF) solution, the Ca/P atomic ratio demonstrated that the apatite formation amount on the coated sample was greater than that on the uncoated nanocomposite.Furthermore, assessing the corrosion resistance indicated that the coatings on the Mg-based substrate led to a corrosion current density(icorr) that was considerably lower than that of the substrate.Such a condition revealed that the coating would provide an obstacle for the corrosion.Based on this study, the PLGA/hardystonite(HT) composite-coated Mg–3Zn–0.5Ag–15NiTi nanocomposite may be suitably applied as an orthopedic implant biomaterial.

A Trojan horse biomimetic delivery system using mesenchymal stem cells for HIF-1α siRNA-loaded nanoparticles on retinal pigment epithelial cells under hypoxia environment

AIM: To demonstrate the feasibility of mesenchymal stem cell(MSC)-mediated nano drug delivery, which was characterized by the “Trojan horse”-like transport of hypoxiainducible factor-1α small interfering RNA(HIF-1α si RNA) between MSCs and retinal pigment epithelial cells(RPE) under hypoxia environment.METHODS: Plasmid and lentivirus targeting the human HIF-1α gene were designed and constructed. HIF-1α si RNA was encapsulated into poly(lactic-co-glycolic acid) nanoparticles(PLGA-NPs) through the water-in-oil-in-water(w/o/w) multiple emulsion technique. The effect of PLGANPs uptake on the expression of HIF-1α m RNA was tested in RPE cells by real-time quantitative polymerase chain reaction(q PCR) and additional transfected conditions were used as control, including lentivirus group, nude plasmid group and blank PLGA group. MSCs were transfected with the NPs and the transfection efficacy was evaluated by flow cytometry. Transwell co-culture system of transfected MSCs and RPE cells was constructed under hypoxia environment. The effects of MSC-loaded HIF-1α si RNA PLGA-NPs on proliferation, apoptosis, and migration of RPE cells were then evaluated. The effect of transfected MSCs on HIF-1α expression of RPE cells was analyzed by using q PCR at the time points 24h, 3d, and 7d.RESULTS: The average diameter of PLGA-NPs loaded with HIF si RNA was 314.1 nm and the zeta potential was-0.36 m V. The transfection efficiency of PLGA-NPs was 67.3%±5.2% into MSCs by using flow cytometry. Compared with the lentivirus group, the PLGA-NPs loaded with HIF-1α si RNA can effectively reduce the expression of HIF-1α m RNA up to 7d in RPE(0.63±0.05 at 7d, P<0.001). In the Transwell co-culture system of transfected MSCs and RPE, the abilities of proliferation(2.34±0.17, 2.40±0.28, 2.47±0.24 at 48h, F=0.23, P=0.80), apoptosis(14.83%±2.43%, 12.94%±2.19%, 12.39%±3.21%;F=0.70, P=0.53) and migration(124.5±7.78, 119.5±5.32, 130±9.89, F=1.33, P=0.33) of the RPE cells had no differences between MSCloaded HIF-1α si RNA PLGA-NPs and other groups. The inhibition of PLGA on the HIF-1α m RNA expression in RPE cells could continue until the 7th day, the level of HIF-1α m RNA was lower than that of other groups(F=171.98, P<0.001). CONCLUSION: The delivery of PLGA-NPs loaded with HIF-1α si RNA carried by MSCs is found to be beneficial temporally for HIF-1α m RNA inhibition in RPE cells under hypoxia environment. The MSC-based bio-mimetic delivery of HIF-1α si RNA nanoparticles is a potential method for therapy against choroidal neovascularization.

Synthesis and Characterization of UPPE-PLGA-rhBMP2 Scaffolds for Bone Regeneration

A novel unsaturated polyphosphoester(UPPE) was devised in our previous research,which is a kind of promising scaffold for improving bone regeneration.However,the polymerization process of UPPE scaffolds was unfavorable,which may adversely affect the bioactivity of osteoinductive molecules added if necessary,such as recombinant human bone morphogenetic protein-2(rhBMP2).The purpose of this study was to build a kind of optimal scaffold named UPPE-PLGA-rhBMP2(UPB) and to investigate the bioactivity of rhBMP2 in this scaffold.Furthermore,the cytotoxicity and biocompatibility of UPB scaffold was assessed in vitro.A W1/O/W2 method was used to fabricate PLGA-rhBMP2 microspheres,and then the microspheres were added to UPPE for synthesizing UPB scaffold.The morphological characters of PLGA-rhBMP2 microspheres and UPB scaffolds were observed under the scanning electron microscopy and laser scanning confocal microscopy.The cumulative release of UPB scaffolds was detected by using ELISA.The cytotoxicity and biocompatibility of UPB scaffolds were evaluated through examining the adsorption and apoptosis of bone marrow stromal cells(bMSCs) seeded on the surface of UPB scaffolds.The bioactivity of rhBMP2 in UPB scaffolds was assessed through measuring the alkaline phosphates(ALP) activity in bMSCs seeded.The results showed that UPB scaffolds sequentially exhibited burst and sustained release of rhBMP2.The cytotoxicity was greatly reduced when the scaffolds were immersed in buffer solution for 2 h.bMSCs attached and grew on the surface of soaked UPB scaffolds,exerting well biocompatibility.The ALP activity of bMSCs seeded was significantly enhanced,indicating that the bioactivity of rhBMP2 remained and still took effect after the unfavorable polymerization process of scaffolds.It was concluded that UPB scaffolds have low cytotoxicity,good biocompatibility and preserve bioactivity of rhBMP2.UPB scaffolds are promising in improving bone regeneration.

Preparation of Tolterodine Metabolite Loaded Biodegradable PLGA Microspheres

Biodegradable polymer poly（lactic-co-glycolic acid）（PLGA） was used to encapsulate the pharmacological activity metabolite of tolterodine by means of O/W emulsion solvent evaporation method via homogenization in the emulsification process. The influences of preparation parameters were investigated. The results indicate that increa- sing PLGA concentration from 15% to 40% made the encapsulation efficiency of 5-hydroxymethyl derivative of tol- terodine（5-HMT） increased from 55.39% to 76.32%, and the particle size increased from 34.33 μm to 70,78 lain. In addition, when homogenization speed increased from 850 r/min to 2300 r/min, both particle size and encapsulation efficiency of microspheres decreased. An increase in the volume of aqueous phase led to higher encapsulation efficiency and bigger particle size. Increasing temperature made encapsulation efficiency and particle size change significantly. While reaction temperature increased from 20 ℃ to 50 ℃, the encapsulation efficiency decreased from 70.44% to 24.07%, and particle size increased from 38.66 μm to 69.38 μm. High reaction temperature（over 40 ℃） may lead to porous surface of microspheres. Porous surface, encapsulation efficiency and particle size influenced on the in vitro release of 5-HMT together.