Tieying Yin

Coronary drug-eluting stents: From design optimization to newer strategies

Compared with early bare-metal stents, drug-eluting stents (DESs) are more effective in treating coronary artery diseases, especially in inhibiting restenosis. However, in-stent restenosis still clinically occurs at a non-negligible rate. More importantly, delayed endothelialization, inflammation, and hypersensitivity trigger subacute or late adverse events, particularly stent thrombosis, and thereby raise more concerns over the long-term safety of DESs. These problems are mostly associated with the permanent polymeric materials, non-optimal therapeutic drugs, and/or metallic stent platforms used in current DES design. It is critically important to further improve and optimize DES design and apply newer strategies for developing next generation DES. These new generation DESs should maintain their clinical efficacy and meanwhile eliminate the long-term safety concerns. In this review article, the current information on the optimization of DES design was critically reviewed based on DESs basic components, namely, stent platform, restenotic drug, and polymer coating. The available strategies for designing next-generation DESs were also summarized, ranging from degradable polymer DESs, to polymer-free DESs, to fully biodegradable DESs.

In vitro and in vivo investigations on the effects of low-density lipoprotein concentration polarization and haemodynamics on atherosclerotic localization in rabbit and zebrafish

Atherosclerosis (AS) commonly occurs in the regions of the arterial tree with haemodynamic peculiarities, including local flow field disturbances, and formation of swirling flow and vortices. The aim of our study was to confirm low-density lipoprotein (LDL) concentration polarization in the vascular system in vitro and in vivo, and investigate the effects of LDL concentration polarization and flow field alterations on atherosclerotic localization. Red fluorescent LDL was injected into optically transparent Flk1: GFP zebrafish embryos, and the LDL distribution in the vascular lumen was investigated in vivo using laser scanning confocal microscopy. LDL concentration at the vascular luminal surface was found to be higher than that in the bulk. The flow field conditions in blood vessel segments were simulated and measured, and obvious flow field disturbances were found in the regions of vascular geometry change. The LDL concentration at the luminal surface of bifurcation was significantly higher than that in the straight segment, possibly owing to the atherogenic effect of disturbed flow. Additionally, a stenosis model of rabbit carotid arteries was generated. Atherosclerotic plaques were found to have occurred in the stenosis group and were more severe in the stenosis group on a high-fat diet. Our findings provide the first ever definite proof that LDL concentration polarization occurs in the vascular system in vivo. Both lipoprotein concentration polarization and flow field changes are involved in the infiltration/accumulation of atherogenic lipids within the location of arterial luminal surface and promote the development of AS.

Study of biocompatibility of medical grade high nitrogen nickel-free austenitic stainless steel in vitro

Adverse effects of nickel ions being released into the living organism have resulted in development of high nitrogen nickel-free austenitic stainless steels for medical applications. Nitrogen not only replaces nickel for austenitic structure stability but also improves steel properties. The cell cytocompatibility, blood compatibility and cell response of high nitrogen nickel-free austenitic stainless steel were studied in vitro. The mechanical properties and microstructure of this stainless steel were compared to the currently used 316L stainless steel. It was shown that the new steel material had comparable basic mechanical properties to 316L stainless steel and preserved the single austenite organization. The cell toxicity test showed no significant toxic side effects for MC3T3-E1 cells compared to nitinol alloy. Cell adhesion testing showed that the number of MC3T3-E1 cells was more than that on nitinol alloy and the cells grew in good condition. The hemolysis rate was lower than the national standard of 5% without influence on platelets. The total intracellular protein content and ALP activity and quantification of mineralization showed good cell response. We conclude that the high nitrogen nickel-free austenitic stainless steel is a promising new biomedical material for coronary stent development.

Re-Endothelialization Study on Endovascular Stents Seeded by Endothelial Cells through Up- or Downregulation of VEGF

We studied the effects of gene transfection of endothelial cells with vascular endothelial growth factor (VEGF) on re-endothelialization and inhibition of in-stent restenosis. Transfected endothelial cells (ECs) exposed to different VEGF levels were seeded on a stent surface for evaluation in vitro. VEGF121(++) ECs and VEGF121(--) ECs were established using lentiviral-mediated HUVECs transfection. VEGF RNA transcription level and VEGF protein expression were detected by qPCR, Western blot, and ELISA. Methyl thiazolyl tetrazolium (MTT) assay, wound healing assay, and in vitro HUVEC tube formation assay showed that VEGF overexpression promoted cell proliferation, migration, and endothelial capillary-like tube formation. Downregulation of VEGF expression inhibited these activities. Using a rotational culturing system, cells tightly adhered on the stent surface. Stents seeded with transfected ECs at different VEGF levels were implanted in abdominal aortas of New Zealand white rabbits to study re-endothelialization and inhibition of in-stent restenosis. Stents with cells exposed to excess VEGF expression were almost completely covered with cells after stent implantation for 1 week (w). In the VEGF interference group this process was delayed over 4 w due to RNAi-mediated silencing of VEGF. Cryosectioning after 12 w showed that stents seeded with HUVECs exposed to excess VEGF expression significantly reduced the neointima area and stenosis when compared with bare metal stents and stents from the VEGF interference group. Transgenic HUVECs were not found in tissues of experimental animals. Furthermore, cells from these tissues were similar to those from normal tissue. In conclusion, VEGF-mediated endothelialization was found. Furthermore, ECs exposed to VEGF overexpression reduced neointimal hyperplasia, promoted endothelialization, and reduced in-stent restenosis.

Endothelialization and in‐stent restenosis on the surface of glycoprotein IIIa monoclonal antibody eluting stent

Since the percutaneous transtuminal coronary angioplasty was introduced into China in 1984, this procedure has become widely accepted as an important step in coronary revascularization. This study shows the effect of the monoclonal antibody (mAb) on the platelet glycoprotein IIIa receptor during endothelialization and in-stent restenosis by implanting the mAb-eluting stents into iliac arteries of rabbits. The hard tissue cross sections of the stent-implanted arterial segments were made by polymethylmethacrylate embedding. Arterial intima proliferation was observed and analyzed. The endothelialization of the stent surface was observed using scanning electron microscope, whereas the ultrastructure of the neointima was observed using transmission electron microscope. After one month of stent implantation, the surfaces of both groups were covered by intact endothelial layers, but the neointimal areas and the ratio of stenosis were significantly lesser in the mAb-eluting stent group (p < 0.01). After 3 months, the ratio of stenosis in the mAb-eluting stent group was 14.67 ± 0.79, whereas that of the bare stent group was 21.58 ± 1.76 (p < 0.01). Therefore, the mAb eluting from the stent surface has the potential to accelerate endothelialization, prevent thrombosis formation due to the interaction of stent with blood, and decrease the stenosis ratio by inhibiting neointima proliferation.

Immobilization of heparin/poly-l-lysine microspheres on medical grade high nitrogen nickel-free austenitic stainless steel surface to improve the biocompatibility and suppress thrombosis

Thrombosis formation, restenosis, and delayed endothelium regeneration continue to be a challenge for coronary artery stent therapy. To improve the hemocompatibility of cardiovascular implants and to selectively direct vascular cell behavior, a novel heparin/poly-l-lysine microsphere was developed and immobilized on a dopamine-coated surface. We chose medical grade high nitrogen nickel-free austenitic stainless steel as the stent material since it has better biocompatibility. The stability and structural characteristics of the microspheres changed with the heparin: poly-l-lysine concentration ratio. Antithrombin III binding was significantly enhanced. Furthermore, for plasma coagulation tests, the activated partial thromboplastin time and thrombin time were prolonged and depended on the heparinfunction. The modified exhibited excellent stability and anticoagulant activity, and efficiently accelerated endothelialization and anticoagulation. This work has potential application for the design of coronary artery stent surfaces tailored for vascular cell behavior.

Non-invasive approaches for drug delivery to the brain based on the receptor mediated transport

The blood brain barrier (BBB) is a physical and biochemical barrier that prevents entry of toxic compounds into brain for preserving homeostasis. However, the BBB also strictly limits influx of most therapeutic agents into the brain. One promising method for overcoming this problem to deliver drugs is receptor mediated transport (RMT) system, which employs the vesicular trafficking machinery to transport substrates across the BBB endothelium in a noninvasive manner. The conjugates of drug or drug-loaded vector linked with appropriate ligands specifically binds to the endogenous targeting receptor on the surface of the endothelial cells. Then drugs could enter the cell body by means of transcytosis and eventual releasing into the brain parenchyma. Over the past 20years, there have been significant developments of RMT targeting strategies. Here, we will review the recent advance of various promising RMT systems and discuss the capability of these approaches for drug delivery to the brain.

Effects of gamma irradiation and moist heat for sterilization on sodium alginate.

Polysaccharides, such as alginates, are already being used as carriers for drug delivery. The physicochemical and biological properties of alginates may be affected via irradiation and thermal treatments. To explore and compare effects of two kinds of sterilization methods, gamma irradiation and moist heat, on sodium alginate (SA), physicochemical and biological properties of SA powder and solutions were investigated after sterilization. Human umbilical vein endothelial cells (HUVEC) was used to assess the cytotoxicity of the SA after sterilization. The research showed that 25 kGy gamma ray can effectively sterilize microorganism. Both gamma irradiation and moist heat hardly affect the native pH of SA. Compared to irradiation sterilization, moist heat sterilization showed smaller changes in intrinsic viscosity for all SA samples and lead to less glycosidic bond breaking of SA powders. The moist heat sterilization can cause the main chain scission and double bonds formation of the SA solutions. Cytotoxicity studies demonstrated that sterilized SA powers and SA solutions treated by gamma ray sterilization can increase the viability of HUVEC. However, SA solutions treated by moist heat sterilization were found to present severe cytotoxicity. The research results may provide interesting future advancements toward the development of SA-based products for biomedical applications.

Eluting characteristics of a platelet glycoprotein receptor antibody using a PLLA-coated stent

Drug-eluting stents (DES) have emerged as a recognized alternative to treat stent restenosis but many questions remain regarding the optimal type and eluting characteristics of both drug and stent. The first component of the study examines the extent of surface coating of PLLA (poly(L-lactic acid)) on a Nitinol stent. The second characterizes the adsorption and elution rates of monoclonal mouse anti-human platelet glycoprotein (GP) IIIa antibody SZ-21 from a PLLA-coated surface. The PLLA coating was examined by fluorescence staining and image analysis using the Image Processing Box of MATLAB. Stents exposed to the monoclonal mouse anti-human platelet GP IIIa antibody were tested for their adsorption characteristics by radioisotope technique with 125I-labelled SZ-21. The elution rates were then measured in looped circuits at different velocities (10 or 20 ml/min) and durations (30 min up to 312 h). Results showed that the fluorescence staining and image analysis showed a striking difference in the extent of coating between PLLA-coated stents and SZ-21 eluting stents on the gray-scale distribution of Nitinol surfaces. The amount of SZ-21 adsorbed onto the PLLA-coated stents was dependent on the concentration and duration of immersion in the solution. The method of preparation the mAb eluting stent significantly influenced the elution characteristics for a continuous perfusion of more than 300 h. The eluting curve was biphasic with initial rapid elution for the first 24 h followed by a gradual slow elution. These results indicate that the Image Processing Box of MATLAB appears to be a useful method for semi-quantitative analysis of fluorescence images. Furthermore, SZ-21 can be passively adsorbed onto PLLA-coated stents and predictably influenced by the concentration and duration of immersion. These studies may pave the way to developing stent-based delivery of a potent anti-platelet agent.

Recent advancements in the use of exosomes as drug delivery systems

Extracellular vesicles (EVs) are the substances that are released by most types of cells and have an important role in cell to cell communication. Among the most highly researched EVs are exosome. Recent studies show that exosomes derived from cells have different roles and targets. Many studies show that exosome can efficiently deliver many different kinds of cargo to the target cell. Therefore, they are often used to deliver therapeutic cargo for treatment. The exosomes that have been used include both natural ones and those that have been modified with other substances to increase the delivery ability. This article provides a review of both exosomes derived from various cells and modified exosome and their ability in delivering the many kinds of cargo to the target cell.