Recent advances in surface functionalization techniques on polymethacrylate materials for optical biosensor applications.

Biosensor chips for immune-based assay systems have been investigated for their application in early diagnostics. The development of such systems strongly depends on the effective protein immobilization on polymer substrates. In order to achieve this complex heterogeneous interaction the polymer surface must be functionalized with chemical groups that are reactive towards proteins in a way that surface functional groups (such as carboxyl, -COOH; amine, -NH2; and hydroxyl, -OH) chemically or physically anchor the proteins to the polymer platform. Since the proteins are very sensitive towards their environment and can easily lose their activity when brought in close proximity to the solid surface, effective surface functionalization and high level of control over surface chemistry present the most important steps in the fabrication of biosensors. This paper reviews recent developments in surface functionalization and preparation of polymethacrylates for protein immobilization. Due to their versatility and cost effectiveness, this particular group of plastic polymers is widely used both in research and in industry.

Utilization of Nitrogen-Doped Graphene Quantum Dots to Neutralize ROS and Modulate Intracellular Antioxidant Pathways to Improve Dry Eye Disease Therapy.

Purpose: Patients afflicted with dry eye disease (DED) experience significant discomfort. The underlying cause of DED is the excessive accumulation of ROS on the ocular surface. Here, we investigated the nitrogen doped-graphene quantum dots (NGQDs), known for their ROS-scavenging capabilities, as a treatment for DED. Methods: NGQDs were prepared by using citric acid and urea as precursors through hydrothermal method. The antioxidant abilities of NGQDs were evaluated through: scavenging the ROS both extracellular and intracellular, regulating the nuclear factor-erythroid 2-related factor (Nrf2) antioxidant pathway of human corneal epithelial cells (HCECs) and their transcription of inflammation related genes. Furthermore, NGQDs were modified by Arg-Gly-Asp-Ser (RGDS) peptides to obtain RGDS@NGQDs. In vivo, both the NGQDs and RGDS@NGQDs were suspended in 0.1% Pluronic F127 (w/v) and delivered as eye drops in the scopolamine hydrobromide-induced DED mouse model. Preclinical efficacy was compared to the healthy and DPBS treated DED mice. Results: These NGQDs demonstrated pronounced antioxidant properties, efficiently neutralizing free radicals and activating the intracellular Nrf2 pathway. In vitro studies revealed that treatment of H2O2-exposed HCECs with NGQDs induced a preservation in cell viability. Additionally, there was a reduction in the transcription of inflammation-associated genes. To prolong the corneal residence time of NGQDs, they were further modified with RGDS peptides and suspended in 0.1% Pluronic F127 (w/v) to create RGDS@NGQDs F127 eye drops. RGDS@NGQDs exhibited superior intracellular antioxidant activity even at low concentrations (10 µg/mL). Subsequent in vivo studies revealed that RGDS@NGQDs F127 eye drops notably mitigated the symptoms of DED mouse model, primarily by reducing ocular ROS levels. Conclusion: Our findings underscore the enhanced antioxidant benefits achieved by modifying GQDs through nitrogen doping and RGDS peptide tethering. Importantly, in a mouse model, our novel eye drops formulation effectively ameliorated DED symptoms, thereby representing a novel therapeutic pathway for DED management.

A tissue-adhesive F127 hydrogel delivers antioxidative copper-selenide nanoparticles for the treatment of dry eye disease.

Dry eye disease (DED) is currently the most prevalent condition seen in ophthalmology outpatient clinics, representing a significant public health issue. The onset and progression of DED are closely associated with oxidative stress-induced inflammation and damage. To address this, an aldehyde-functionalized F127 (AF127) hydrogel eye drop delivering multifunctional antioxidant Cu2-xSe nanoparticles (Cu2-xSe NPs) was designed. The research findings revealed that the Cu2-xSe nanoparticles exhibit unexpected capabilities in acting as superoxide dismutase and glutathione peroxidase. Additionally, Cu2-xSe NPs possess remarkable efficacy in scavenging reactive oxygen species (ROS) and mitigating oxidative damage. Cu2-xSe NPs displayed promising therapeutic effects in a mouse model of dry eye. Detailed investigation revealed that the nanoparticles exert antioxidant, anti-apoptotic, and inflammation-mitigating effects by modulating the NRF2 and p38 MAPK signalling pathways. The AF127 hydrogel eye drops exhibit good adherence to the ocular surface through the formation of Schiff-base bonds. These findings suggest that incorporating antioxidant Cu2-xSe nanoparticles into a tissue-adhesive hydrogel could present a highly effective therapeutic strategy for treating dry eye disease and other disorders associated with reactive oxygen species. STATEMENT OF SIGNIFICANCE: A new formulation for therapeutic eye drops to be used in the treatment of dry eye disease (DED) was developed. The formulation combines copper-selenium nanoparticles (Cu2-xSe NPs) with aldehyde-functionalized Pluronic F127 (AF127). This is the first study to directly examine the effects of Cu2-xSe NPs in ophthalmology. The NPs exhibited antioxidant capabilities and enzyme-like properties. They effectively eliminated reactive oxygen species (ROS) and inhibited apoptosis through the NRF2 and p38 MAPK signalling pathways. Additionally, the AF127 hydrogel enhanced tissue adhesion by forming Schiff-base links. In mouse model of DED, the Cu2-xSe NPs@AF127 eye drops demonstrated remarkable efficacy in alleviating symptoms of DED. These findings indicate the potential of Cu2-xSe NPs as a readily available and user-friendly medication for the management of DED.

Polymeric Microspheres Designed to Carry Crystalline Drugs at Their Surface or Inside Cavities and Dimples.

Injectable polymer microparticles with the ability to carry and release pharmacologically active agents are attracting more and more interest. This study is focused on the chemical synthesis, characterization, and preliminary exploration of the utility of a new type of injectable drug-releasing polymer microparticle. The particles feature a new combination of structural and physico-chemical properties: (i) their geometry deviates from the spherical in the sense that the particles have a cavity; (ii) the particles are porous and can therefore be loaded with crystalline drug formulations; drug crystals can reside at both the particle's surfaces and inside cavities; (iii) the particles are relatively dense since the polymer network contains covalently bound iodine (approximately 10% by mass); this renders the drug-loaded particles traceable (localizable) by X-ray fluoroscopy. This study presents several examples. First, the particles were loaded with crystalline voriconazole, which is a potent antifungal drug used in ophthalmology to treat fungal keratitis (infection/inflammation of the cornea caused by penetrating fungus). Drug loading as high as 10% by mass (=mass of immobilized drug/(mass of the microparticle + mass of immobilized drug) × 100%) could be achieved. Slow local release of voriconazole from these particles was observed in vitro. These findings hold promise regarding new approaches to treat fungal keratitis. Moreover, this study can help to expand the scope of the transarterial chemoembolization (TACE) technique since it enables the use of higher drug loadings (thus enabling higher local drug concentration or extended therapy duration), as well as application of hydrophobic drugs that cannot be used in combination with existing TACE embolic particles.

Sustained Release of Voriconazole Using 3D-Crosslinked Hydrogel Rings and Rods for Use in Corneal Drug Delivery.

Corneal disorders and diseases are prevalent in the field of clinical ophthalmology. Fungal keratitis, one of the major factors leading to visual impairment and blindness worldwide, presents significant challenges for traditional topical eye drop treatments. The objective of this study was to create biocompatible 3D-crosslinked hydrogels for drug delivery to the cornea, intending to enhance the bioavailability of ophthalmic drugs. Firstly, a series of flexible and porous hydrogels were synthesized (free-radical polymerization), characterized, and evaluated. The materials were prepared by the free-radical polymerization reaction of 1-vinyl-2-pyrrolidinone (also known as N-vinylpyrrolidone or NVP) and 1,6-hexanediol dimethacrylate (crosslinker) in the presence of polyethylene glycol 1000 (PEG-1000) as the porogen. After the physicochemical characterization of these materials, the chosen hydrogel demonstrated outstanding cytocompatibility in vitro. Subsequently, the selected porous hydrogels could be loaded with voriconazole, an antifungal medication. The procedure was adapted to realize a loading of 175 mg voriconazole per ring, which slightly exceeds the amount of voriconazole that is instilled into the eye via drop therapy (a single eye drop corresponds with approximately 100 mg voriconazole). The voriconazole-loaded rings exhibited a stable zero-order release pattern over the first two hours, which points to a significantly improved bioavailability of the drug. Ex vivo experiments using the established porcine eye model provided confirmation of a 10-fold increase in drug penetration into the cornea (after 2 h of application of the hydrogel ring, 35.8 ± 3.2% of the original dose is retrieved from the cornea, which compares with 3.9 ± 1% of the original dose in the case of eye drop therapy). These innovative hydrogel rods and rings show great potential for improving the bioavailability of ophthalmic drugs, which could potentially lead to reduced hospitalization durations and treatment expenses.

A novel approach to achieve semi-sustained drug delivery to the eye through asymmetric loading of soft contact lenses.

Soft contact lenses are increasingly being explored as a vehicle for controlled delivery of ophthalmic drugs. However, traditional methods of drug-loading by soaking have limitations such as burst delivery and the release of drugs at the front side of the lens, leading to poor drug efficacy and systemic side effects. This study introduces a new methodology, termed asymmetric drug loading, whereby the ophthalmic drug 'Rebamipide' is attached to and released from the post-lens (=cornea-contacting) surface exclusively. The methodology involves using polymeric microparticles that carry a lipophilic crystalline ophthalmic drug at their surface. These drug-loaded microparticles first transfer the drug to the concave surface of the contact lens, and when worn, the drug is transferred again, now from the lens to the cornea. This is achieved through the diffusion of the drug from one hydrophobic microenvironment (the silicone moieties of the contact lens polymer network) to another hydrophobic microenvironment (the corneal epithelium) over a short pathway. The second drug transfer was observed and studied in experiments using an ex vivo porcine eye model. The results show that the drug amount that was absorbed by the cornea after applying the rebamipide-loaded contact lenses is approximately 3× (10.7 ± 3.1 µg) as much as the amount of rebamipide that gets transferred after the instillation of one eye drop (1% solution (p < 0.001). The new drug-loading method offers a practical and reproducible means of delivering ophthalmic drugs to the cornea through soft contact lenses. The drug payloads achieved are comparable to dosages used during eye drop therapy.

Versatile polymer microspheres for injection therapy: aspects of fluoroscopic traceability and biofunctionalization.

Synthesis and characterization of a series of novel microspheres featuring (i) radiopacity (i.e., clear fluoroscopic traceability) and (ii) an outer surface exposing aldehyde groups are reported. The aldehydes allowed us to tether proteins onto the particles' surface under mild conditions, under which the protein conformation and, hence, structural motifs for biorecognition are preserved. Essential monomer building blocks were (i) 4-iodobenzoyl-2-oxo-ethylmethacrylate (4-IEMA) for radiopacity and (ii) propenal for surface tethering of proteins. The particles demonstrated good X-ray visibility and cytocompatibility. Procedures to couple proteins onto the surface were optimized using fluorescent bovine serum albumin (FITC-BSA) or collagen (FITC-collagen). Furthermore, radiopaque microparticles with unlabeled bovine collagen type I were produced. The presence of immobilized collagen was verified with narrow-scan X-ray photoelectron spectroscopy. Fibroblasts readily adhere to and grow on the collagen-modified surfaces, whereas this was much less the case for the unmodified controls. The results led us to suggest that immobilized nondenatured collagen may transform filler particles from passive space-occupying objects to particles that cross-talk with surrounding tissues.

Synthesis and characterization of radiopaque iodine-containing degradable PVA hydrogels.

Poly (vinyl alcohol) (PVA) hydrogels are highly attractive for biomedical applications, especially for controlled release of drugs and proteins. Recently, degradable PVA hydrogels have been described, having the advantage that the material disappears over time from the implantation site. Herein, we report the synthesis of radiopaque degradable PVA, which gives a further advantage that the position of the hydrogel can precisely be determined by X-ray fluoroscopy. Radiopacity has been introduced by replacing 0.5% of the pendent alcohol groups on the PVA with 4-iodobenzoylchloride. This level of substitution rendered the polymer adequately radiopaque. The subsequent modification of 0.8% of the pendent hydroxyl groups with an ester acrylate functional group allowed for cross-linking of the macromers. The radiopaque hydrogels degraded over a time span of 140 days. Rheology data suggested that the macromer solutions were appropriate for injection.

New intrinsically radiopaque hydrophilic microspheres for embolization: synthesis and characterization.

Polymeric particles currently used for embolization procedures have the disadvantage that they are radiolucent, that is, invisible on X-ray images, and consequently the interventional radiologist has to resort to angiography to (indirectly) monitor the fate of the particles. Here, we introduce intrinsically radiopaque hydrophilic microspheres. Since these microspheres can directly be visualized on X-ray images, using these microspheres for embolization purposes will allow superprecise location of the embolic material, both during and after the procedure. The microspheres, which are prepared by suspension polymerization, are based on the radiopaque monomer 2-[4-iodobenzoyl]-oxo-ethylmethacrylate and hydroxyethylmethacrylate (HEMA) and/or 1-vinyl-2-pyrrolidinone (NVP) as hydrophilic component. It has been shown that for clinically relevant X-ray visibility the spheres should contain at least 20 wt % iodine. At this iodine content, copolymerization with HEMA results in spheres that hardly imbibe water (EQ = 1.08). When HEMA is replaced by NVP, the volume swelling ratio can be significantly increased (to 1.33).

New acrylic microspheres for arterial embolization: combining radiopacity for precise localization with immobilized thrombin to trigger local blood coagulation.

Particles currently used in arterial embolization therapy have several disadvantages, most importantly their radiolucency. This means the radiologist cannot precisely asses the fate of embolization particles. Microspheres that combine two additional features have been designed. By incorporating an iodine containing monomer, radiopaque microspheres were obtained that display good visibility under standard X-ray conditions. Incorporation of methacrylic acid makes the surface of the spheres suitable for surface functionalization. Here, thrombin was covalently attached to the surface of the radiopaque microspheres. By induction of a thrombus, improved anchoring of the embolization spheres in the blood vessel can be obtained. The immobilized thrombin induced a biphasic response of the blood namely: (1) fast deposition of fibrin on the surface resulting in sphere aggregation and (2) additional thrombin generation in the surrounding blood and a subsequent local thrombus formation. These microspheres with both intrinsic X-ray visibility and a biofunctionalized surface can potentially improve embolization therapies.

Thrombus formation at the surface of guide-wire models: effects of heparin-releasing or heparin-exposing surface coatings.

PURPOSE: This study was conducted to investigate whether thrombus formation at the surface of guide wires occurs, and--if so--whether this can be suppressed or prevented through incorporation of heparin in the surface coating. MATERIALS AND METHODS: Five guide wire models were examined; three had a polymeric hydrophilic surface coating (90/10 guide wire), which was either heparin-free, impregnated with sodium-heparin (Na-hep), or impregnated with benzalkonium heparin (BAK-hep). The other two guide wires had a coating of polytetrafluoroethylene (PTFE), either without heparin, or impregnated with BAK-hep. Release of heparin, exposure of heparin at the surface of the guide wires, thrombogenicity (under static and flow conditions) and their propensity to attract blood platelets were investigated. RESULTS: The guide wire 90/10 Na-hep releases approximately 150-200 mU active heparin per cm coil within the first few minutes after incubation in buffer. The PTFE BAK-hep shows a relatively slow release of 60-70 mU active heparin per cm coil. The 90/10 BAK-hep showed no released heparin but the most exposed heparin. In a static experiment with human full blood excessive thrombus formation occurred at the heparin-free models, whereas the others remained essentially clean. In a thrombin-generation assay under flow the authors observed strong retardation of thrombin formation in the case of the 90/10 Na-hep guide wire. CONCLUSIONS: The static and dynamic in vitro assays, taken together, show that the 90/10 Na-hep provides a coating with an extremely low level of surface thrombogenicity. Use of a guide wire with a hydrophilic distal coating that releases and exposes sodium heparin may contribute to the safety of diagnostic and therapeutic interventional procedures.

Polymers with tunable toxicity: a reference scale for cytotoxicity testing of biomaterial surfaces.

A series of copolymers, with varying ratio di-methylamino-ethylmethacrylate (DMAEMA) and methyl-methacrylate (MMA), was designed as a potential scale for cytotoxicity. These copolymers were characterized for toxicity of their surface. The surfaces of washed copolymers display increasing toxicity with increasing DMAEMA content. The toxicity was observed for three different cell-types, namely mouse fibroblasts, human endothelial cells and human osteoblast-like cells. With an increasing toxic surface, cell growth was inhibited as was indicated by the proliferation marker Ki-67. Staining for F-actin revealed that with increasing DMAEMA, cells adopted a more and more round morphology, resulting in decreased surface-contact area. Immuno-staining for phospho-tyrosine or vinculin demonstrated gradual loss of focal adhesions on increasingly toxic surfaces. Surprisingly loss of focal adhesions coincided with an increase in paxillin and vinculin protein, indicating cells try compensating for loss of adhesion. This series of copolymers may have potential as a cytotoxicity scale. They provoke cellular responses ranging from highly toxic to completely non-toxic, with some showing intermediate toxicity.

Capacity and tolerance of a new device for ocular drug delivery.

A new method to increase the drug-capacity of the OphthaCoil, a flexible and tubular device for delivery of drugs to the tear film of the eye, was explored. Poly(2-hydroxyethyl methacrylate)- and poly(2-hydroxyethyl methacrylate-co-1-vinyl-2-pyrrolidone)-microspheres were prepared by suspension polymerization. The resultant particles were swollen in a highly concentrated solution of either the dye fluorescein sodium or the antibiotic chloramphenicol. The loaded particles were placed in the central cavity of the ocular device. In vitro release profiles showed a six-fold increase of the capacity for the dye fluorescein sodium, but not for the antibiotic chloramphenicol. Flexibility measurements revealed that by introducing microspheres in the central cavity of the device, flexibility did not decrease. Finally, a preliminary in vivo evaluation of the device (n=5) was done for a 2h-period to assess the tolerance of the device in the human eye. Ophthalmologic examinations and photographs of the eye indicated no signs of irritation. Volunteers reported that the presence of the device in the eye could be noticed, but no irritation was reported.

Towards a functional radiopaque hydrogel for nucleus pulposus replacement.

Patients with severe back pain, attributed to a herniation of the nucleus pulposus of the intervertebral disc, can benefit from a replacement of only the nucleus pulposus, provided the annulus fibrosus is still functional. This study investigated four intrinsically radiopaque hydrogel biomaterials, which were designed specifically to replace the herniated nucleus pulposus. The important characteristic of these hydrogels is that they can be visualized entirely with both MRI and X-rays. The materials are based on copolymers of N-vinyl-2-pyrrolidinone (NVP) or 2-hydroxyethyl methacrylate (HEMA) and a radiopacity introducing monomer, 2-(4'-iodobenzoyl)-oxo-ethyl methacrylate (4IEMA). Two of the formulations also contain the chemical crosslinker allyl methacrylate (AMA). Physical-mechanical properties like the water-uptake, biocompatibility, stiffness, and fatigue and creep behavior were studied, while keeping an eye on the intended application. All four materials were designed with 5-6 mass % of iodine to ensure sufficient X-ray visibility between two vertebrae. It was found that the materials display appropriate stiffness and biocompatibility. The crosslinked materials hold most promise as a functional nucleus prosthesis, as they combine these properties also with high water content, fatigue resistance, and recovery after loading.

Biocompatibility of a new radiopaque iodine-containing acrylic bone cement.

Radiopacity in the vast majority of the commercially available acrylic bone cements that are used clinically is provided by particles of either BaSO(4) or ZrO(2). Literature reports have shown these agents to have a detrimental effect on some mechanical properties of the cements as well as on its biological response. We, therefore, have developed a new type of bone cement, for which radiopacity results from the presence of an iodine-containing methacrylic copolymer. The focus of the present work was the comparison of the biocompatibility of this new cement and a commercially available cement that contains barium sulfate. In vitro experiments show that both cements are cytocompatible materials, for which no toxic leachables are found. Implantation of the cements in a rabbit for three months resulted in the occasional presence of a thin fibrous tissue at the cement-bone interface, which is common for acrylic bone cements. Consideration of all the results led to the conclusion that the new cement is as biocompatible as the BaSO(4)-containing one.

The effect of high-density-lipoprotein on thrombus formation on and endothelial cell attachement to biomaterial surfaces.

Cardiovascular implants such as vascular grafts fail frequently because they lack genuine blood-compatibility. The blood-contacting surface should simultaneously prevent thrombus formation and promote formation of a confluent endothelial cell layer, to achieve sustained haemostasis. Contact activation and endothelialization are known to be determined by the plasma proteins which adsorb onto virtually all synthetic surfaces almost immediately upon contact with blood. A common approach in blood-compatibility research is, therefore, to use hydrophilic biomaterials, which are sometimes claimed to be "protein-repellent". We report here that, for synthetic polymeric surfaces, hydrophilicity is by no means synonymous to protein-repellency. We discovered that significant amounts of proteins, especially high-density lipoprotein, adsorb to hydrophilic surfaces. Pre-incubation of hydrophilic synthetic surfaces with high-density lipoprotein provides a blood-biomaterial interface, which inhibits thrombin generation and subsequent thrombus formation, and also accommodates overgrowth with a confluent endothelial layer. This approach may open the way to truly functional small-caliber arterial prostheses, and may also be relevant to cardiovascular tissue engineering in which de novo vascular tissues are cultured on or within a biomaterial scaffold.

In vitro and in vivo evaluation of drug-eluting microspheres designed for transarterial chemoembolization therapy.

Poly(D,L-lactic acid) biodegradable microspheres, loaded with the drugs cisplatin and/or sorafenib tosylate, were prepared, characterized and studied. Degradation of the microspheres, and release of cisplatin and/or sorafenib tosylate from them, were investigated in detail. Incubation of the drug-carrying microspheres in phosphate buffered saline (pH=7.4) revealed slow degradation. Nevertheless, significant release of cisplatin and sorafenib tosylate from microspheres loaded with both drugs was apparent in vitro; this can be attributed to their porous structure. Supernatants from microspheres loaded with both drugs showed strong toxic effects on cells (i.e. endothelial cells, fibroblast cells and Renca tumor cells) and potent anti-angiogenic effect in the matrigel endothelial tube assay. In vivo anti-tumor effects of the microspheres were also observed, in a Renca tumor mouse model. The poly(D,L-lactic acid) microspheres containing both cisplatin and sorafenib tosylate revealed highest therapeutic efficacy, probably demonstrating that combined local administration of cisplatin and sorafenib tosylate synergistically inhibits tumor growth in situ. In conclusion, this study demonstrates the applicability of biodegradable poly(D,L-lactic acid) microspheres loaded with cisplatin and sorafenib tosylate for local drug delivery as well as the potential of these microspheres for future use in transarterial chemoembolization.

Synthesis and characterization of a new vertebroplasty cement based on gold-containing PMMA microspheres.

There are a number of drawbacks to incorporating large concentrations of barium sulfate (BaSO4) as the radiopacifier in PMMA-based bone cements for percutaneous vertebroplasty. These include adverse effects on injectability, viscosity profile, setting time, mechanical properties of the cement and bone resorption. We have synthesized a novel cement that is designed to address some of these drawbacks. Its powder includes PMMA microspheres in which gold particles are embedded and its monomer is the same as that used in commercial cements for vertebroplasty. In comparison to one such commercial cement brand, VertaPlex™, the new cement has longer doughing time, longer injection time, higher compressive strength, higher compressive modulus, and is superior in terms of cytotoxicity. For augmentation of fractured fresh-frozen cadaveric vertebral bodies (T6-L5) using simulated vertebroplasty, results for compressive strength and compressive stiffness of the construct and the percentage of the volume of the vertebral body filled by the cement were comparable for the two cements although the radiopacity of the new cement was significantly lower than that for VertaPlex™. The present results indicate that the new cement warrants further study.

Intrinsically radiopaque hydrogels for nucleus pulposus replacement.

Degeneration of the intervertebral disc is the most common cause of back pain. In case of early stage degenerative disc disease or traumatic herniations, a suitable treatment may be to replace the nucleus pulposus, preserving the annulus fibrosus. Eight new hydrogel biomaterials were prepared and studied for their potential as a nucleus replacement. The hydrogels were designed according to the following criteria: (i), they should exhibit adequate radiopacity; (ii), they should be non-cytotoxic; (iii), implantation in the dry state and subsequent swelling in situ to fill the entire nucleus cavity; (iv), after swelling they should match the physical-mechanical properties of the native nucleus. The approach was to use copolymers consisting of 2-(4'-iodobenzoyl)-oxo-ethyl methacrylate (4IEMA) and a hydrophilic building block (either N-vinyl-2-pyrrolidinone (NVP) or 2-hydroxyethyl methacrylate (HEMA)); 4 copolymers of NVP/4IEMA and 4 copolymers of HEMA/4IEMA in different compositions (5, 10, 15 and 20 mol% 4IEMA). The study comprised 1H-NMR analysis of the copolymerization reaction NVP+4IEMA. Furthermore, the copolymers were studied with respect to their swelling behavior, mechanical properties, cytotoxicity in vitro and X-ray contrast. Hydrogels with 5 mol% 4IEMA appear to meet all criteria: they are non-cytotoxic, have adequate physical-mechanical properties and feature sufficient radiopacity in a realistic model. The potential implications of these new results with respect to treatment of degenerative disc disease are discussed briefly.

Hydrophobicity as a design criterion for polymer scaffolds in bone tissue engineering.

Porous polymeric scaffolds play a key role in most tissue-engineering strategies. A series of non-degrading porous scaffolds was prepared, based on bulk-copolymerisation of 1-vinyl-2-pyrrolidinone (NVP) and n-butyl methacrylate (BMA), followed by a particulate-leaching step to generate porosity. Biocompatibility of these scaffolds was evaluated in vitro and in vivo. Furthermore, the scaffold materials were studied using the so-called demineralised bone matrix (DBM) as an evaluation system in vivo. The DBM, which is essentially a part of a rat femoral bone after processing with mineral acid, provides a suitable environment for ectopic bone formation, provided that the cavity of the DBM is filled with bone marrow prior to subcutaneous implantation in the thoracic region of rats. Various scaffold materials, differing with respect to composition and, hence, hydrophilicity, were introduced into the centre of DBMs. The ends were closed with rat bone marrow, and ectopic bone formation was monitored after 4, 6, and 8 weeks, both through X-ray microradiography and histology. The 50:50 scaffold particles were found to readily accommodate formation of bone tissue within their pores, whereas this was much less the case for the more hydrophilic 70:30 counterpart scaffolds. New healthy bone tissue was encountered inside the pores of the 50:50 scaffold material, not only at the periphery of the constructs but also in the center. Active osteoblast cells were found at the bone-biomaterial interfaces. These data indicate that the hydrophobicity of the biomaterial is, most likely, an important design criterion for polymeric scaffolds which should promote the healing of bone defects. Furthermore, it is argued that stable, non-degrading porous biomaterials, like those used in this study, provide an important tool to expand our comprehension of the role of biomaterials in scaffold-based tissue engineering approaches.