Hicham Fenniri

Nanotechnology-based drug delivery systems

Nanoparticles hold tremendous potential as an effective drug delivery system. In this review we discussed recent developments in nanotechnology for drug delivery. To overcome the problems of gene and drug delivery, nanotechnology has gained interest in recent years. Nanosystems with different compositions and biological properties have been extensively investigated for drug and gene delivery applications. To achieve efficient drug delivery it is important to understand the interactions of nanomaterials with the biological environment, targeting cell-surface receptors, drug release, multiple drug administration, stability of therapeutic agents and molecular mechanisms of cell signalling involved in pathobiology of the disease under consideration. Several anti-cancer drugs including paclitaxel, doxorubicin, 5-fluorouracil and dexamethasone have been successfully formulated using nanomaterials. Quantom dots, chitosan, Polylactic/glycolic acid (PLGA) and PLGA-based nanoparticles have also been used for in vitro RNAi delivery. Brain cancer is one of the most difficult malignancies to detect and treat mainly because of the difficulty in getting imaging and therapeutic agents past the blood-brain barrier and into the brain. Anti-cancer drugs such as loperamide and doxorubicin bound to nanomaterials have been shown to cross the intact blood-brain barrier and released at therapeutic concentrations in the brain. The use of nanomaterials including peptide-based nanotubes to target the vascular endothelial growth factor (VEGF) receptor and cell adhesion molecules like integrins, cadherins and selectins, is a new approach to control disease progression.

Widespread nanoparticle-assay interference: implications for nanotoxicity testing.

The evaluation of engineered nanomaterial safety has been hindered by conflicting reports demonstrating differential degrees of toxicity with the same nanoparticles. The unique properties of these materials increase the likelihood that they will interfere with analytical techniques, which may contribute to this phenomenon. We tested the potential for: 1) nanoparticle intrinsic fluorescence/absorbance, 2) interactions between nanoparticles and assay components, and 3) the effects of adding both nanoparticles and analytes to an assay, to interfere with the accurate assessment of toxicity. Silicon, cadmium selenide, titanium dioxide, and helical rosette nanotubes each affected at least one of the six assays tested, resulting in either substantial over- or under-estimations of toxicity. Simulation of realistic assay conditions revealed that interference could not be predicted solely by interactions between nanoparticles and assay components. Moreover, the nature and degree of interference cannot be predicted solely based on our current understanding of nanomaterial behaviour. A literature survey indicated that ca. 95% of papers from 2010 using biochemical techniques to assess nanotoxicity did not account for potential interference of nanoparticles, and this number had not substantially improved in 2012. We provide guidance on avoiding and/or controlling for such interference to improve the accuracy of nanotoxicity assessments.

Arginine-glycine-aspartic acid modified rosette nanotube-hydrogel composites for bone tissue engineering.

An RGDSK (Arg-Gly-Asp-Ser-Lys) modified rosette nanotube (RNT) hydrogel composite with unique surface chemistry and favorable cytocompatibility properties for bone repair was developed and investigated. The RNTs are biologically inspired nanomaterials obtained through the self-assembly of a DNA base analog (G wedge C base) with tailorable chemical functionality and physical properties. In this study, a cell-adhesive RGDSK peptide was covalently attached to the G wedge C base, assembled into RNTs, and structurally characterized by (1)H/(13)C NMR spectroscopy, mass spectrometry, and electron microscopy. Importantly, results showed that the RGDSK modified RNT hydrogels caused around a 200% increase in osteoblast (bone-forming cell) adhesion relative to hydrogel controls. In addition, osteoblast proliferation was enhanced on RNT hydrogels compared to hydrogel controls after 3 days, which further confirmed the promising cytocompatibility properties of this scaffold. When analyzing the mechanism of increased osteoblast density on RNT hydrogels, it was found that more fibronectin (a protein which promotes osteoblast adhesion) adsorption occurred on RNT coated hydrogels than uncoated hydrogels. As osteoblast adhesion was greatly enhanced on RNT coated hydrogels compared to poly l-lysine and collagen coated hydrogels, this study indicated that not only the surface chemistry was important in improving osteoblast density (via lysine or RGD groups functionalized on RNTs), but also the biomimetic nanoscale properties of RNTs provided a cell-favorable environment. These results warrant further studies on RNTs in hydrogels for better bone tissue regeneration.

Biologically inspired rosette nanotubes and nanocrystalline hydroxyapatite hydrogel nanocomposites as improved bone substitutes

Today, bone diseases such as bone fractures, osteoporosis and bone cancer represent a common and significant public health problem. The design of biomimetic bone tissue engineering materials that could restore and improve damaged bone tissues provides exciting opportunities to solve the numerous problems associated with traditional orthopedic implants. Therefore, the objective of this in vitro study was to create a biomimetic orthopedic hydrogel nanocomposite based on the self-assembly properties of helical rosette nanotubes (HRNs), the osteoconductive properties of nanocrystalline hydroxyapatite (HA), and the biocompatible properties of hydrogels (specifically, poly(2-hydroxyethyl methacrylate), pHEMA). HRNs are self-assembled nanomaterials that are formed from synthetic DNA base analogs in water to mimic the helical nanostructure of collagen in bone. In this study, different geometries of nanocrystalline HA were controlled by either hydrothermal or sintering methods. 2 and 10 wt% nanocrystalline HA particles were well dispersed into HRN hydrogels using ultrasonication. The nanocrystalline HA and nanocrystalline HA/HRN hydrogels were characterized by x-ray diffraction, transmission electron microscopy, and scanning electron microscopy. Mechanical testing studies revealed that the well dispersed nanocrystalline HA in HRN hydrogels possessed improved mechanical properties compared to hydrogel controls. In addition, the results of this study provided the first evidence that the combination of either 2 or 10 wt% nanocrystalline HA and 0.01 mg ml(-1) HRNs in hydrogels greatly increased osteoblast (bone-forming cell) adhesion up to 236% compared to hydrogel controls. Moreover, this study showed that HRNs stimulated HA nucleation and mineralization along their main axis in a way that is very reminiscent of the HA/collagen assembly pattern in natural bone. In summary, the presently observed excellent properties of the biomimetic nanocrystalline HA/HRN hydrogel composites make them promising candidates for further study for bone tissue engineering applications.

Helical rosette nanotubes: a more effective orthopaedic implant material

Due to the nanometric properties of some physiological components of bone, nanomaterials have been proposed as the next generation of improved orthopaedic implant materials. Yet current efforts in the design of orthopaedic materials such as titanium (Ti) are not aimed at tailoring their nanoscale features, which is now believed to be one reason why Ti sometimes fails clinically as a bone implant material. Much effort is thus being dedicated to developing improved bioactive nanometric surfaces and nanomaterials for biospecificity. Helical rosette nanotubes (HRN) are a new class of self-assembled organic nanotubes possessing biologically-inspired nanoscale dimensions. Because of their chemical and structural similarity with naturally-occurring nanostructured constituent components in bone such as collagen and hydroxyapatite, we anticipated that an HRN-coated surface may simulate an environment that bone cells are accustomed to interacting with. The objective of the present in vitro study is therefore to determine the efficacy of HRN as a bone prosthetic material. Results of this study clearly show that both HRN-K1 and HRN-Arg coated Ti displayed enhanced cell adhesion when compared to uncoated Ti. Enhanced cell adhesion was observed even at concentrations as low as 0.005 mg ml−1. These results point towards new possibilities in bone tissue engineering as they serve as a starting point for further mechanistic studies as well as future manipulation of the outer chemistries of HRN to improve the results beyond those presented here. One such effort is the incorporation of peptide sequences on the outer surface of HRN and/or growth factors known to enhance bone functions.

Enhanced Osteoblast Adhesion on Self-Assembled Nanostructured Hydrogel Scaffolds

The objective of the current in vitro study was to improve properties of a commonly used hydrogel for implant applications by incorporating novel self-assembled helical rosette nanotubes (HRNs). Since traditional methods (such as autografts and allografts) used to treat bone defects present various disadvantages (such as donor tissue shortage, extensive inflammation, possible disease transmission, and poor new bone growth), which may lead to implant failure, much effort has been devoted to creating a novel bone substitute that biomimics the nanoscale features of natural bone in order to improve bone growth. HRNs (formed by chemically immobilizing two DNA base pairs) are a novel type of soft nanomaterial that biomimics natural nanostructured components of bone (such as collagen) since they are 3.5 nm in diameter and self-assemble into a helical structure in aqueous solutions. Because HRNs undergo a phase transition from a liquid to a viscous gel when heated to slightly above body temperatures or when added directly to serum-supplemented or serum-free media at body temperatures, they may provide an exciting therapy to heal bone fractures in situ. In this study, HRN-K1 (HRNs functionalized with lysine amino acids) was embedded in and coated on a model hydrogel [specifically, poly(2-hydroxyethyl methacrylate) or pHEMA]. The results of this study showed, for the first time, enhanced osteoblast (bone-forming cell) adhesion on HRN-K1 embedded in and coated on hydrogels compared to hydrogels without HRN-K1. Moreover, the results showed that embedding HRN-K1 into hydrogels can greatly decrease the polymerization time of pHEMA (especially at low temperatures). The presence of lysine in HRN-K1/hydrogels was shown to be one, but not only, property of HRN-K1 that enhanced osteoblast adhesion. In summary, the present results demonstrated that HRNs can improve properties of one particular hydrogel (pHEMA) and, thus, should be further investigated as a bone-healing material.

Biomimetic helical rosette nanotubes and nanocrystalline hydroxyapatite coatings on titanium for improving orthopedic implants.

Natural bone consists of hard nanostructured hydroxyapatite (HA) in a nanostructured protein-based soft hydrogel template (ie, mostly collagen). For this reason, nanostructured HA has been an intriguing coating material on traditionally used titanium for improving orthopedic applications. In addition, helical rosette nanotubes (HRNs), newly developed materials which form through the self-assembly process of DNA base pair building blocks in body solutions, are soft nanotubes with a helical architecture that mimics natural collagen. Thus, the objective of this in vitro study was for the first time to combine the promising attributes of HRNs and nanocrystalline HA on titanium and assess osteoblast (bone-forming cell) functions. Different sizes of nanocrystalline HA were synthesized in this study through a wet chemical precipitation process following either hydrothermal treatment or sintering. Transmission electron microscopy images showed that HRNs aligned with nanocrystalline HA, which indicates a high affinity between both components. Some of the nanocrystalline HA formed dense coatings with HRNs on titanium. More importantly, results demonstrated enhanced osteoblast adhesion on the HRN/nanocrystalline HA-coated titanium compared with conventional uncoated titanium. Among all the HRN/nanocrystalline HA coatings tested, osteoblast adhesion was the greatest when HA nanometer particle size was the smallest. In this manner, this study demonstrated for the first time that biomimetic HRN/nanocrystalline HA coatings on titanium were cytocompatible for osteoblasts and, thus, should be further studied for improving orthopedic implants.

Enhanced endothelial cell functions on rosette nanotube-coated titanium vascular stents

One of the main problems with current vascular stents is a lack of endothelial cell interactions, which if sufficient, would create a uniform healthy endothelium masking the underlying foreign metal from inflammatory cell interference. Moreover, if endothelial cells from the arterial wall do not adhere to the stent, the stent can become loose and dislodge. Therefore, the objective of this in vitro study was to design a novel biomimetic nanostructured coating (that does not contain drugs) on conventional vascular stent materials (specifically, titanium) for improving vascular stent applications. Rosette nanotubes (RNTs) are a new class of biomimetic nanotubes that self-assemble from DNA base analogs and have been shown in previous studies to sufficiently coat titanium and enhance osteoblast cell functions. RNTs have many desirable properties for use as vascular stent coatings including spontaneous self-assembly in body fluids, tailorable surface chemistry for specific implant applications, and nanoscale dimensions similar to those of the natural vascular extracellular matrix. Importantly, the results of this study provided the first evidence that RNTs functionalized with lysine (RNT–K), even at low concentrations, significantly increase endothelial cell density over uncoated titanium. Specifically, 0.01 mg/mL RNT–K coated titanium increased endothelial cell density by 37% and 52% compared to uncoated titanium after 4 h and three days, respectively. The excellent cytocompatibility properties of RNTs (as demonstrated here for the first time for endothelial cells) suggest the need for the further exploration of these novel nanostructured materials for vascular stent applications.

Self-assembled rosette nanotubes encapsulate and slowly release dexamethasone.

Rosette nanotubes (RNTs) are novel, self-assembled, biomimetic, synthetic drug delivery materials suitable for numerous medical applications. Because of their amphiphilic character and hollow architecture, RNTs can be used to encapsulate and deliver hydrophobic drugs otherwise difficult to deliver in biological systems. Another advantage of using RNTs for drug delivery is their biocompatibility, low cytotoxicity, and their ability to engender a favorable, biologically-inspired environment for cell adhesion and growth. In this study, a method to incorporate dexamethasone (DEX, an inflammatory and a bone growth promoting steroid) into RNTs was developed. The drug-loaded RNTs were characterized using diffusion ordered nuclear magnetic resonance spectroscopy (DOSY NMR) and UV-Vis spectroscopy. Results showed for the first time that DEX can be easily and quickly encapsulated into RNTs and released to promote osteoblast (bone-forming cell) functions over long periods of time. As a result, RNTs are presented as a novel material for the targeted delivery of hydrophobic drugs otherwise difficult to deliver.

Water-Soluble J-Type Rosette Nanotubes with Giant Molar Ellipticity

A new self-assembling tricyclic module (×K1) featuring the Watson-Crick H-bonding arrays of guanine and cytosine fused to an internal pyridine ring was synthesized. When dissolved in water at room temperature, this module rapidly self-assembles into hexameric rosettes, which then stack to form J-type rosette nanotubes (RNTs) with increased inner/outer diameters and the largest molar ellipticity ever reported (4 × 10(6) deg·M(-1)·m(-1)). Using a combination of imaging and spectroscopic techniques we established the structure of ×K1-RNT and have shown that the extended π system of the self-assembling module resulted in a new family of J-type RNTs with enhanced intermodular electronic communication.