Suitability of Marine- and Porcine-Derived Collagen Type I Hydrogels for Bioprinting and Tissue Engineering Scaffolds.

Collagens from a wide array of animals have been explored for use in tissue engineering in an effort to replicate the native extracellular environment of the body. Marine-derived biomaterials offer promise over their conventional mammalian counterparts due to lower risk of disease transfer as well as being compatible with more religious and ethical groups within society. Here, collagen type I derived from a marine source (Macruronus novaezelandiae, Blue Grenadier) is compared with the more established porcine collagen type I and its potential in tissue engineering examined. Both collagens were methacrylated, to allow for UV crosslinking during extrusion 3D printing. The materials were shown to be highly cytocompatible with L929 fibroblasts. The mechanical properties of the marine-derived collagen were generally lower than those of the porcine-derived collagen; however, the Young's modulus for both collagens was shown to be tunable over a wide range. The marine-derived collagen was seen to be a potential biomaterial in tissue engineering; however, this may be limited due to its lower thermal stability at which point it degrades to gelatin.

Design, Synthesis and Characterization of a Novel Type of Thermo-Responsible Phospholipid Microcapsule-Alginate Composite Hydrogel for Drug Delivery.

Liposomes are extensively used in drug delivery, while alginates are widely used in tissue engineering. However, liposomes are usually thermally unstable and drug-leaking when in liquids, while the drug carriers made of alginates show low loading capacities when used for drug delivery. Herein, we developed a type of thermo-responsible liposome-alginate composite hydrogel (TSPMAH) by grafting thermo-responsive liposomes onto alginates by using Ca2+ mediated bonding between the phosphatidic serine (PS) in the liposome membrane and the alginate. The temperature-sensitivity of the liposomes was actualized by using phospholipids comprising dipalmitoylphosphatidylcholine (DPPC) and PS and the liposomes were prepared by a thin-film dispersion method. The TSPMAH was then successfully prepared by bridge-linking the microcapsules onto the alginate hydrogel via PS-Ca2+-Carboxyl-alginate interaction. Characterizations of the TSPMAH were carried out using scanning electron microscopy, transform infrared spectroscopy, and laser scanning confocal microscopy, respectively. Their rheological property was also characterized by using a rheometer. Cytotoxicity evaluations of the TSPMAH showed that the composite hydrogel was biocompatible, safe, and non-toxic. Further, loading and thermos-inducible release of model drugs encapsulated by the TSPMAH as a drug carrier system was also studied by making protamine-siRNA complex-carrying TSPMAH drug carriers. Our results indicated that the TSPMAH described herein has great potentials to be further developed into an intelligent drug delivery system.

Advances in printing biomaterials and living cells: implications for islet cell transplantation.

PURPOSE OF REVIEW: Rapid advances in bioprinting have attracted the attention of the clinical world. The advent of printable, cytocompatible materials and appropriate hardware provides an unprecedented ability to design and create 3D structures throughout which living cells and bioactive components are strategically distributed. Here, we review those advances and present how they can be used to create new structures for more effective islet cell transplantation. RECENT FINDINGS: There is a need for improvements in the delivery vehicle for transplantable islet cells. Significant progress has been made in 3D printing of multicellular structures and vascularized structures and multiple bioactive molecules. Strategies for extending these recent findings to islet transplantation are discussed. More importantly, the first promising step, 3D printing human islets has recently been demonstrated. SUMMARY: The advent of 3D bioprinting provides unprecedented opportunities for islet transplantation. Highlighting the capabilities of 3D bioprinting should also encourage clinicians to consider other areas appropriate for its use.

Bipolar Electrochemical Stimulation Using Conducting Polymers for Wireless Electroceuticals and Future Directions.

Electrochemistry has become a powerful strategy to modulate cellular behavior and biological activity by manipulating electrical signals. Subsequent electrical stimulus-responsive conducting polymers (CPs) have advanced traditional wired electrochemical stimulation (ES) systems and developed wireless cell stimulation systems due to their electroconductivity, biocompatibility, stability, and flexibility. Bipolar electrochemistry (BPE), i.e., wireless electrochemistry, offers an effective pathway to modify wired ES systems into a desirable contactless mode, turning out a potential technique to offer fundamental insights into neural cell stimulation and neural network formation. This review commences with a brief discussion of the BPE technique and also the advantages of a bipolar electrochemical stimulation (BPES) system compared to traditional wired ES systems and other wireless ES systems. Then, the BPES system is elucidated through four aspects: the benefits of BPES, the key factors to establish BPES platforms for cell stimulation, the limits/barriers to overcome for current rigid materials in particular metals-based systems, and a brief overview of the concept proved by CPs-based systems. Furthermore, how to refine the existing BPES system from materials/devices modification that combine CP compositions with 3D fabrication/bioprinting technologies is elaborately discussed as well. Finally, the review ends together with future research directions, picturing the potential of BPES system in biomedical applications.

Fibrinogen, collagen, and transferrin adsorption to poly(3,4-ethylenedioxythiophene)-xylorhamno-uronic glycan composite conducting polymer biomaterials for wound healing applications.

We present the conducting polymer poly (3,4-ethylenedioxythiophene) (PEDOT) doped with an algal-derived glycan extract, Phycotrix™ [xylorhamno-uronic glycan (XRU84)], as an innovative electrically conductive material capable of providing beneficial biological and electrical cues for the promotion of favorable wound healing processes. Increased loading of the algal XRU84 into PEDOT resulted in a reduced surface nanoroughness and interfacial surface area and an increased static water contact angle. PEDOT-XRU84 films demonstrated good electrical stability and charge storage capacity and a reduced impedance relative to the control gold electrode. A quartz crystal microbalance with dissipation monitoring study of protein adsorption (transferrin, fibrinogen, and collagen) showed that collagen adsorption increased significantly with increased XRU84 loading, while transferrin adsorption was significantly reduced. The viscoelastic properties of adsorbed protein, characterized using the ΔD/Δf ratio, showed that for transferrin and fibrinogen, a rigid, dehydrated layer was formed at low XRU84 loadings. Cell studies using human dermal fibroblasts demonstrated excellent cell viability, with fluorescent staining of the cell cytoskeleton illustrating all polymers to present excellent cell adhesion and spreading after 24 h.

Shaping collagen for engineering hard tissues: Towards a printomics approach.

Hard tissue engineering has evolved over the past decades, with multiple approaches being explored and developed. Despite the rapid development and success of advanced 3D cell culture, 3D printing technologies and material developments, a gold standard approach to engineering and regenerating hard tissue substitutes such as bone, dentin and cementum, has not yet been realised. One such strategy that differs from conventional regenerative medicine approach of other tissues, is the in vitro mineralisation of collagen templates in the absence of cells. Collagen is the most abundant protein within the human body and forms the basis of all hard tissues. Once mineralised, collagen provides important support and protection to humans, for example in the case of bone tissue. Multiple in vitro fabrication strategies and mineralisation approaches have been developed and their success in facilitating mineral deposition on collagen to achieve bone-like scaffolds evaluated. Critical to the success of such fabrication and biomineralisation approaches is the collagen template, and its chemical composition, organisation, and density. The key factors that influence such properties are the collagen processing and fabrication techniques utilised to create the template, and the mineralisation strategy employed to deposit mineral on and throughout the templates. However, despite its importance, relatively little attention has been placed on these two critical factors. Here, we critically examine the processing, fabrication and mineralisation strategies that have been used to mineralise collagen templates, and offer insights and perspectives on the most promising strategies for creating mineralised collagen scaffolds. STATEMENT OF SIGNIFICANCE: In this review, we highlight the critical need to fabricate collagen templates with advanced processing techniques, in a manner that achieves biomimicry of the hierarchical collagen structure, prior to utilising in vitro mineralisation strategies. To this end, we focus on the initial collagen that is selected, the extraction techniques used and the native fibril forming potential retained to create reconstituted collagen scaffolds. This review synthesises current best practises in material sourcing, processing, mineralisation strategies and fabrication techniques, and offers insights into how these can best be exploited in future studies to successfully mineralise collagen templates.

Hybrid Printing Using Cellulose Nanocrystals Reinforced GelMA/HAMA Hydrogels for Improved Structural Integration.

3D printing of soft-tissue like cytocompatible single material constructs with appropriate mechanical properties remains a challenge. Hybrid printing technology provides an attractive alternative as it combines a cell-free ink for providing mechanical support with a bioink for housing embedded cells. Several hybrid printed structures have been developed, utilizing thermoplastic polymers such as polycaprolactone as structural support. These thermoplastics demonstrated limited structural integration with the cell-laden components, and this may compromise the overall performance. In this work, a hybrid printing platform is presented using two distinct hydrogel inks that share the same photo-crosslinking chemistry to enable simple fabrication and seamless structural integration. A mechanically reinforced hydrogel ink is developed comprising cellulose nanocrystals and gelatin methacryloyl/hyaluronic acid methacrylate (GelMA/HAMA) as the structural component, and GelMA/HAMA as the cytogel containing a mouse chondrogenic cell line, ATDC5. Hybrid printed constructs with encapsulated cells are fabricated using the two optimized inks, and the structural integration of the constructs is evaluated by cyclic mechanical compression. Finally, the cell viability of encapsulated ATDC5 cells in the hybrid printed structures is evaluated.

3D hybrid printing platform for auricular cartilage reconstruction.

As scaffolds approach dimensions that are of clinical relevance, mechanical integrity and distribution becomes an important factor to the overall success of the implant. Hydrogels often lack the structural integrity and mechanical properties for use in vivo or handling. The inclusion of a structural support during the printing process, referred to as hybrid printing, allows the implant to retain structure and protect cells during maturation without needing to compromise its biological performance. In this study, scaffolds for the purpose of auricular cartilage reconstruction were evaluated via a hybrid printing approach using methacrylated Gelatin (GelMA) and Hyaluronic acid (HAMA) as the cell-laden hydrogel, Polycaprolactone (PCL) as structural support and Lutrol F-127 as sacrificial material. Furthermore, printing parameters such as nozzle diameter, strand spacing and filament orientation scaffolds were investigated. Compression and bending tests showed that increasing nozzle sizes decrease the compressive modulus of printed scaffolds, with up to 82% decrease in modulus when comparing between a 400 µm and 200 µm sized nozzle tip at the same strand spacing. On the contrary, strand spacing and orientation influences mainly the bending modulus due to the greater porosity and changes in pore size area. Using a 400 µm sized nozzle, scaffolds fabricated have a measured compression and bending modulus in the range similar to the native cartilage. The viability and proliferation of human mesenchymal stem cells delivered within the bioink was not affected by the printing process. Using results obtained from mechanical testing, a scaffold with matching mechanical properties across six distinct regions mimicking the human auricular cartilage can be completed in one single print process. The use of PCL and GelMA-HAMA as structural support and cell-laden hydrogel respectively are an excellent combination to provide tailored mechanical integrity, while maintaining porosity and protection to cells during differentiation.

Electrofluidic control of bioactive molecule delivery into soft tissue models based on gelatin methacryloyl hydrogels using threads and surgical sutures.

The delivery of bioactive molecules (drugs) with control over spatial distribution remains a challenge. Herein, we demonstrate for the first time an electrofluidic approach to controlled delivery into soft tissue models based on gelatin methacryloyl (GelMA) hydrogels. This was achieved using a surgical suture, whereby transport of bioactive molecules, including drugs and proteins, was controlled by imposition of an electric field. Commonly employed surgical sutures or acrylic threads were integrated through the hydrogels to facilitate the directed introduction of bioactive species. The platform consisted of two reservoirs into which the ends of the thread were immersed. The anode and cathode were placed separately into each reservoir. The thread was taken from one reservoir to the other through the gel. When current was applied, biomolecules loaded onto the thread were directed into the gel. Under the same conditions, the rate of movement of the biomolecules along GelMA was dependent on the magnitude of the current. Using 5% GelMA and a current of 100 µA, 2 uL of fluorescein travelled through the hydrogel at a constant velocity of 7.17 ± 0.50 um/s and took less than 8 minutes to exit on the thread. Small molecules such as riboflavin migrated faster (5.99 ± 0.40 µm/s) than larger molecules such as dextran (2.26 ± 0.55 µm/s with 4 kDa) or BSA (0.33 ± 0.07 µm/s with 66.5 kDa). A number of commercial surgical sutures were tested and found to accommodate the controlled movement of biomolecules. Polyester, polyglactin 910, glycolide/lactide copolymer and polyglycolic acid braided sutures created adequate fluid connection between the electrodes and the hydrogel. With a view to application in skin inflammatory diseases and wound treatment, wound healing, slow and controlled delivery of dexamethasone 21-phosphate disodium salt (DSP), an anti-inflammatory prodrug, was achieved using medical surgicryl PGA absorbable suture. After 2 hours of electrical stimulation, still 81.1% of the drug loaded was encapsulated within the hydrogel.

A simple technique for development of fibres with programmable microsphere concentration gradients for local protein delivery.

Alginate has been a biologically viable option for controlled local delivery of bioactive molecules in vitro and in vivo. Specific bioactive molecule release profiles are achieved often by controlling polymer composition/concentration, which also determines the modulus of hydrogels. This largely limits alginate-mediated bioactive molecule delivery to single-factors of uniform concentration applications, rather than applications that may require (multiple) bioactive molecules delivered at a concentration gradient for chemotactic purposes. Here we report a two-phase PLGA/alginate delivery system composed of protein-laden poly-d,l-lactic-co-glycolic acid (PLGA) microspheres wet-spun into alginate fibres. Fluorescein isothiocyanate-conjugated bovine serum albumin (FITC-BSA) was used as a model protein and the developed structures were characterized. The fabrication system devised was shown to produce wet-spun fibres with a protein concentration gradient (G-Alg/PLGA fibre). The two-phase delivery matrices display retarded FITC-BSA release in both initial and late stages compared to release from the PLGA microspheres or alginate fibre alone. In addition, incorporation of higher concentrations of protein-loaded PLGA microspheres increased protein release compared to the fibres with lower concentrations of BSA-loaded microspheres. The "programmable" microsphere concentration gradient fibre methodology presented here may enable development of novel alginate scaffolds with the ability to guide tissue regeneration through tightly-controlled release of one or more proteins in highly defined spatio-temporal configurations.

Development of rhamnose-rich hydrogels based on sulfated xylorhamno-uronic acid toward wound healing applications.

An array of biological properties is demonstrated in the category of extracts broadly known as ulvans, including antibacterial, anti-inflammatory and anti-coagulant activities. However, the development of this category in biomedical applications is limited due to high structural variability across species and a lack of consistent and scalable sources. In addition, the modification and formulation of these molecules is still in its infancy with regard to progressing to product development. Here, a sulfated and rhamnose-rich, xylorhamno-uronic acid (XRU) extract from the cell wall of a controlled source of cultivated Australian ulvacean macroalgae resembles mammalian connective glycosaminoglycans. It is therefore a strong candidate for applications in wound healing and tissue regeneration. This study targets the development of polysaccharide modification for fabrication of 3D scaffolds for skin cell (fibroblast) culture. The XRU extract is methacrylated and UV-crosslinked to produce hydrogels with tuneable mechanical properties. The hydrogels demonstrate high cell viability and support cell proliferation over 14 days, which are far more functional than comparable alginate gels. Importantly, an XRU-based bioink is developed for extrusion printing 3D constructs both with and without cell encapsulation. These results highlight the close to product potential of this rhamnose-rich XRU extract as a promising biomaterial toward wound healing. Future studies should be focused on in-depth in vitro characterizations to examine the role of the material in dermal extracellular matrix (ECM) secretion of 3D printed structures, and in vivo characterizations to assess its capacity in supporting wound healing.

On Low-Concentration Inks Formulated by Nanocellulose Assisted with Gelatin Methacrylate (GelMA) for 3D Printing toward Wound Healing Application.

Cellulose nanofibrils (CNFs) in the form of hydrogels stand out as a platform biomaterial in bioink formulation for 3D printing because of their low cytotoxicity and structural similarity to extracellular matrices. In the present study, 3D scaffolds were successfully printed with low-concentration inks formulated by 1 w/v % 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-oxidized CNF with less than 1 w/v % gelatin methacrylate (GelMA). Quartz crystal microbalance with dissipation monitoring (QCM-D) measurements showed strong interaction between the two biopolymers. The UV cross-linking ability of GelMA (≤1 w/v %) was enhanced in the presence of TEMPO-oxidized CNFs. Multiple factors including strong physical interaction between CNF and GelMA, in situ cross-linking of CNF by Ca2+, and UV cross-linking of GelMA enabled successful 3D printing of low-concentration inks of CNF/GelMA into scaffolds possessing good structural stability. The mechanical strength of the scaffolds was tuned in the range of 2.5 to 5 kPa. The cell culture with 3T3 fibroblasts revealed noncytotoxic and biocompatible features for the formulated inks and printed scaffolds. More importantly, the incorporated GelMA in the CNF hydrogel promoted the proliferation of fibroblasts. The developed low-concentration CNF/GelMA formulations with a facile yet effective approach to fabricate scaffolds showed great potential in 3D printing for wound healing application.

Development of a Coaxial 3D Printing Platform for Biofabrication of Implantable Islet-Containing Constructs.

Over the last two decades, pancreatic islet transplantations have become a promising treatment for Type I diabetes. However, although providing a consistent and sustained exogenous insulin supply, there are a number of limitations hindering the widespread application of this approach. These include the lack of sufficient vasculature and allogeneic immune attacks after transplantation, which both contribute to poor cell survival rates. Here, these issues are addressed using a biofabrication approach. An alginate/gelatin-based bioink formulation is optimized for islet and islet-related cell encapsulation and 3D printing. In addition, a custom-designed coaxial printer is developed for 3D printing of multicellular islet-containing constructs. In this work, the ability to fabricate 3D constructs with precise control over the distribution of multiple cell types is demonstrated. In addition, it is shown that the viability of pancreatic islets is well maintained after the 3D printing process. Taken together, these results represent the first step toward an improved vehicle for islet transplantation and a potential novel strategy to treat Type I diabetes.

Aqueous solution behaviour and membrane disruptive activity of pH-responsive PEGylated pseudo-peptides and their intracellular distribution.

The effect of PEGylation on the aqueous solution properties and cell membrane disruptive activity of a pH-responsive pseudo-peptide, poly(l-lysine iso-phthalamide), has been investigated by dynamic light scattering, haemolysis and lactate dehydrogenase (LDH) assays. Intracellular trafficking of the polymers has been examined using confocal and fluorescence microscopy. With increasing degree of PEGylation, the modified polymers can form stabilised compact structures with reduced mean hydrodynamic diameters. Poly(l-lysine iso-phthalamide) with a low degree of PEGylation (17.4 wt%) retained pH-dependent solution behaviour and showed enhanced kinetic membrane disruptive activity compared to the parent polymer. It facilitated trafficking of endocytosed materials into the cytoplasm of HeLa cells. At levels of PEGylation in excess of 25.6 wt%, the modified polymers displayed a single particle size distribution unresponsive to pH, as well as a decrease in cell membrane lytic ability. The mechanism involved in membrane destabilisation was also investigated, and the potential applications of these modified polymers in drug delivery were discussed.

Advanced fabrication approaches to controlled delivery systems for epilepsy treatment.

INTRODUCTION: Epilepsy is a chronic brain disease characterized by unprovoked seizures, which can have severe consequences including loss of awareness and death. Currently, 30% of epileptic patients do not receive adequate seizure alleviation from oral routes of medication. Over the last decade, local drug delivery to the focal area of the brain where the seizure originates has emerged as a potential alternative and may be achieved through the fabrication of drug-loaded polymeric implants for controlled on-site delivery. AREAS COVERED: This review presents an overview of the latest advanced fabrication techniques for controlled drug delivery systems for refractory epilepsy treatment. Recent advances in the different techniques are highlighted and the limitations of the respective techniques are discussed. EXPERT OPINION: Advances in biofabrication technologies are expected to enable a new paradigm of local drug delivery systems through offering high versatility in controlling drug release profiles, personalized customization and multi-drug incorporation. Tackling some of the current issues with advanced fabrication methods, including adhering to GMP-standards and industrial scale-up, together with innovative solutions for complex designs will see to the maturation of these techniques and result in increased clinical research into implant-based epilepsy treatment. ABBREVIATIONS: GMP: Good manufacturing process; DDS(s): Drug delivery system(s); 3D: Three-dimensional; AEDs: Anti-epileptic drugs; BBB: Blood brain barrier; PLA: Polylactic acid; PLGA: Poly(lactic-co-glycolic acid); PCL: poly(ɛ-caprolactone); ESE: Emulsification solvent evaporation; O/W: Oil-in-water; W/O/W: Water-in-oil-in-water; DZP: Diazepam; PHT: Phenytoin; PHBV: Poly(hydroxybutyrate-hydroxyvalerate); PEG: Polyethylene glycol; SWD: Spike-and-wave discharges; CAD: Computer aided design; FDM: Fused deposition modeling; ABS: Acrylonitrile butadiene styren; eEVA: Ethylene-vinyl acetate; GelMA: Gelatin methacrylate; PVA: Poly-vinyl alcohol; PDMS: Polydimethylsiloxane; SLA: Stereolithography; SLS: Selective laser sintering.

Development of drug-loaded polymer microcapsules for treatment of epilepsy.

Despite significant progress in developing new drugs for seizure control, epilepsy still affects 1% of the global population and is drug-resistant in more than 30% of cases. To improve the therapeutic efficacy of epilepsy medication, a promising approach is to deliver anti-epilepsy drugs directly to affected brain areas using local drug delivery systems. The drug delivery systems must meet a number of criteria, including high drug loading efficiency, biodegradability, neuro-cytocompatibility and predictable drug release profiles. Here we report the development of fibre- and sphere-based microcapsules that exhibit controllable uniform morphologies and drug release profiles as predicted by mathematical modelling. Importantly, both forms of fabricated microcapsules are compatible with human brain derived neural stem cells and differentiated neurons and neuroglia, indicating clinical compliance for neural implantation and therapeutic drug delivery.

Evaluation of the Biocompatibility of Polypyrrole Implanted Subdurally in GAERS.

This blinded controlled prospective randomized study investigates the biocompatibility of polypyrrole (PPy) polymer that will be used for intracranial triggered release of anti-epileptic drugs (AEDs). Three by three millimeters PPy are implanted subdurally in six adult female genetic absence epilepsy rats from Strasbourg. Each rat has a polymer implanted on one side of the cortex and a sham craniotomy performed on the other side. After a period of seven weeks, rats are euthanized and parallel series of coronal sections are cut throughout the implant site. Four series of 15 sections are histological (hematoxylin and eosin) and immunohistochemically (neuron-specific nuclear protein, glial fibrillary acidic protein, and anti-CD68 antibody) stained and evaluated by three investigators. The results show that implanted PPy mats do not induce obvious inflammation, trauma, gliosis, and neuronal toxicity. Therefore the authors conclude the PPy used offer good histocompatibility with central nervous system cells and that PPy sheets can be used as intracranial, AED delivery implant.

Modulation of the pH-responsive properties of poly(L-lysine iso-phthalamide) grafted with a poly(ethylene glycol) analogue.

A pH responsive pseudopeptide, poly(L-lysine iso-phthalamide), has been modified with a hydrophilic poly(ethylene glycol) analogue, Jeffamine M-1000 and the effect of grafting ratio on the pH responsive behaviour of the grafted polymers in aqueous solution investigated using fluorescence and 1H NMR spectroscopy. It was demonstrated that at below 35.1 wt% grafting, the modified polymers retained the pH-driven conformational transition of the parent polymer from an expanded structure at high degrees of ionisation to a compact hydrophobically stabilised structure at low degrees of ionisation. The onset of pH response and the pH range over which the conformational transition occurred varied significantly with degree of grafting. At Jeffamine M-1000 ratios in excess of 48.0 wt%, the graft polymer existed in a micellular form over the whole pH studied. Potential applications in drug delivery of both the linear and micellular forms are discussed.

Modulation of cell membrane disruption by pH-responsive pseudo-peptides through grafting with hydrophilic side chains.

The effect of grafting an amphiphilic pseudo-peptide, poly (L-lysine iso-phthalamide), with poly (ethylene glycol) or a hydrophilic poly (ethylene glycol) analogue, Jeffamine M-1000, on the pH-dependent erythrolytic activity and in vitro cytotoxicity have been studied together with the concentration-dependent haemolysis of the polymers with different degrees of grafting. PEGylated polymers showed pH-dependent membrane-disruptive ability similar to the parent poly (L-lysine iso-phthalamide). The polymers showed a better ability to haemolyse the erythrocyte membrane at mildly acidic pHs with increasing degree of PEGylation (up to 17.0 wt.%). Further increasing the degree of PEGylation resulted in a decrease in haemolytic ability. Grafting poly (L-lysine iso-phthalamide) with the lower molecular weight Jeffamine M-1000 had little effect on the haemolytic ability. Finally, the in vitro cytotoxicity of the grafted polymers was assessed by MTT assay, LDH assay and viable cell counts. At pH 7.4, these polymers were well tolerated by a range of mammalian cell lines and grafting reduced the cytotoxicity of polymers. However, at pH 5.5, relative to poly (L-lysine iso-phthalamide), the grafted polymers displayed a better ability to rupture the outer membranes of these cells.

Injectable phenytoin loaded polymeric microspheres for the control of temporal lobe epilepsy in rats.

PURPOSE: Epilepsy is a prevalent neurological disorder with a high frequency of drug resistance. While significant advancements have been made in drug delivery systems to overcome anti-epileptic drug resistance, efficacies of materials in biological systems have been poorly studied. The purpose of the study was to evaluate the anti-epileptic effects of injectable poly(epsilon-caprolactone) (PCL) microspheres for controlled release of an anticonvulsant, phenytoin (PHT), in an animal model of epilepsy. METHODS: PHT-PCL and Blank-PCL microspheres formulated using an oil-in-water (O/W) emulsion solvent evaporation method were evaluated for particle size, encapsulation efficiency, surface morphology and in-vitro drug release profile. Microspheres with the most suitable morphology and release characteristics weresubsequently injected into the hippocampus of a rat tetanus toxin model of temporal lobe epilepsy. Electrocorticography (ECoG)from the cerebral cortex were recorded for all animals. The number of seizure events, severity of seizures, and seizure duration were then compared between the two treatment groups. RESULTS: We have shown that small injections of drug-loaded microspheres are biologically tolerated and released PHT can control seizures for the expected period of time that is in accord with in-vitro release data. CONCLUSION: The study demonstrated the feasibility of polymer-based delivery systems incontrolling focal seizures.