Jennifer J. Kang-Mieler

Advances in ocular drug delivery: emphasis on the posterior segment

Introduction: Recent advances in pharmacological therapies to treat ocular diseases such as glaucoma, age-related macular degeneration, diabetic macular edema and retinal vascular occlusions have greatly improved the prognosis for these diseases. Due to these advances in pharmacological therapy, there is a great deal of interest in minimally invasive delivery methods, which has generated rapid developments in the field of ocular drug delivery. Areas covered: This review will summarize currently available and recent developments for ocular drug delivery to both the anterior and posterior segments. Modes of delivery, including topical, systemic, transcleral/periocular and intravitreal, will be discussed and corresponding examples will be given. This review will highlight the advantages and disadvantages of each mode of delivery and discuss strategies to address these issues. Expert opinion: An ideal therapy should maintain effective levels of drug for the intended duration of treatment following a single application, yet a significant number of months of therapy may be required. There are numerous approaches under investigation to improve treatment options. From the use of novel biomaterial implants and depots for sustained release, to prodrug formations, to iontophoresis to improve drug delivery, the main emphasis will continue to be placed on less invasive, longer acting, sustained release formulations in the treatment of numerous ocular disorders.

Investigation of lysine acrylate containing poly(N‐isopropylacrylamide) hydrogels as wound dressings in normal and infected wounds

The design of materials for cutaneous wound dressings has advanced from passive wound covers to bioactive materials that promote skin regeneration and prevent infection. Crosslinked poly(N-isopropylacrylamide) (PNIPAAm)-based hydrogels have been investigated for a number of biomedical applications. While these materials can be used for drug delivery, limited cell interactions restrict their biological activity. In this article, acryoyl-lysine (A-Lys) was incorporated into poly(ethylene glycol) crosslinked PNIPAAm to enhance biological activity. A-Lys could be incorporated into the hydrogels to improve cellular interaction in vitro, while maintaining swelling properties and thermoresponsive behavior. Polyhexamethylene biguanide, an antimicrobial agent, could be encapsulated and released from the hydrogels and resulted in decreased bacteria counts within 2 hours. Two in vivo animal wound models were used to evaluate the hydrogel wound dressing. First, application of the hydrogels to a rodent cutaneous wound healing model resulted in significant increase in healing rate when compared with controls. Moreover, the hydrogels were also able to decrease bacteria levels in an infected wound model. These results suggest that PNIPAAm hydrogels containing A-Lys are promising wound dressings due to their ability to promote healing and deliver active antimicrobial drugs to inhibit infection.

Decreased Circulation in the Feline Choriocapillaris Underlying Retinal Photocoagulation Lesions

PURPOSE To investigate the effects of argon laser photocoagulation on the choroidal circulation in cats. METHODS Three sizes of argon laser lesions designed to damage the outer retina were created in six cats: larger than 1 mm, 500 μm, and 200 μm. At least 1 month after the lesions, damage to the choroidal vasculature was studied in two ways. First, scanning laser ophthalmoscopy was used to obtain infrared reflectance (IR) photographs and indocyanine green (ICG) angiograms. Second, fluorescent microspheres (15 μm) were injected into the left ventricle. The globes were fixed, the choroid was flat mounted, and images were taken with a fluorescence microscope. Retinal histology was assessed in comparable lesions. RESULTS Histology showed that the inner retina was preserved, but the choroid, tapetum, and outer retina were damaged. ICG angiograms revealed choriocapillaris loss in large lesions and in some 500-μm lesions, whereas the larger vessels were preserved; in 200 μm lesions, choriocapillaris loss was not detectable. However, in all lesions, the distribution of microspheres revealed little if any choriocapillaris flow. In larger lesions, the damaged region was surrounded by an area in which the number of microspheres was higher than in the lesion but lower than in the normal retina. CONCLUSIONS Under lesions that destroyed photoreceptors, the choriocapillaris was also compromised, even when no change could be detected with ICG angiography. Panretinal photocoagulation is designed to increase retinal PO2 by allowing choroidal oxygen to reach the inner retina, but its effectiveness may be limited by damage to the choriocapillaris.

Controlled and Extended In Vitro Release of Bioactive Anti-Vascular Endothelial Growth Factors from a Microsphere-Hydrogel Drug Delivery System

ABSTRACT Purpose: To demonstrate controlled and extended release of bioactive anti-vascular endothelial growth factor (VEGF) agents (ranibizumab or aflibercept) from an injectable microsphere-hydrogel drug delivery system (DDS). Methods: Anti-VEGF agents were radiolabeled with iodine-125 and loaded into poly(lactic-co-glycolic acid) (PLGA) 75:25 microspheres using a modified double-emulsion, solvent evaporation technique. Microspheres were then suspended in an injectable poly(N-isopropylacrylamide)-based thermo-responsive hydrogel to create a microsphere-hydrogel DDS. Release profiles were performed in phosphate buffered saline at 37°C and at predetermined intervals, release samples were collected. Microspheres were also made using non-radiolabeled anti-VEGFs to determine the bioactivity of the DDS throughout release. Bioactivity and cytotoxicity of release samples were determined using human umbilical vascular endothelial cells (HUVECs) under VEGF-induced proliferation. Results: The DDS is capable of releasing either ranibizumab or aflibercept for 196 days with an initial burst (first 24 h) of 22.2 ± 2.2 and 13.1 ± 0.5 μg, respectively, followed by controlled release of 0.153 and 0.065 μg/day, respectively. Release samples showed no toxicity in HUVECs at any time. Both anti-VEGFs remained bioactive throughout release with significant inhibition of HUVEC proliferation compared to the drug-free DDS, which showed no inhibitory effect on HUVEC proliferation. Conclusions: Controlled, extended, and bioactive release for approximately 200 days was achieved for both ranibizumab and aflibercept in vitro. The use of anti-VEGF-loaded microspheres suspended within an injectable, thermo-responsive hydrogel may be an advantageous ocular DDS with the potential to improve upon current therapies.

Extended ocular drug delivery systems for the anterior and posterior segments: biomaterial options and applications

ABSTRACT Introduction: The development of new therapies for treating various eye conditions has led to a demand for extended release delivery systems, which would lessen the need for frequent application while still achieving therapeutic drug levels in the target tissues. Areas covered: Following an overview of the different ocular drug delivery modalities, this article surveys the biomaterials used to develop sustained release drug delivery systems. Microspheres, nanospheres, liposomes, hydrogels, and composite systems are discussed in terms of their primary materials. The advantages and disadvantages of each drug delivery system are discussed for various applications. Recommendations for modifications and strategies for improvements to these basic systems are also discussed. Expert opinion: An ideal sustained release drug delivery system should be able to encapsulate and deliver the necessary drug to the target tissues at a therapeutic level without any detriment to the drug. Drug encapsulation should be as high as possible to minimize loss and unless it is specifically desired, the initial burst of drug release should be kept to a minimum. By modifying various biomaterials, it is possible to achieve sustained drug delivery to both the anterior and posterior segments of the eye.

Objective area measurement technique for choroidal neovascularization from fluorescein angiography

The purpose of this study was to develop a non-biased method of quantitatively measuring choroidal neovascularization (CNV) areas based on late-phase fluorescein angiography (FA) images. Experimental CNV was induced in Long Evans rats by laser disruption of the Bruchs membrane. FA was performed weekly for 5weeks. Multi-Otsu thresholding (MOT) was used to quantify CNV in late-phase FA images from both experimental rodent CNV and wet age-related macular degeneration (wAMD) patients. Images were automatically thresholded into three levels based on the image histogram, with the highest level containing CNV. To determine the techniques ability to quantify CNV areas, rats were given either triamcinolone acetonide or dexamethasone sodium phosphate to treat CNV and compared to untreated rats. The rat CNV lesion areas measured from 5-week histology sections from each treatment group were compared to areas measured from the corresponding FA images. MOT was able to detect statistical decreases in rodent CNV area in the treatment groups versus control from weeks 3 through 5. The ratio of CNV area measured from histology to area measured from FA images was not statistically different between groups. Finally, to determine the usefulness of MOT on pathological morphologies of CNV, MOT was performed on late-phase FA images from patients with classic and diffuse CNV. The technique was able to segment classical CNV in wAMD patients, but performed poorly with diffuse CNV. MOT provides a robust, objective, and quantifiable area measurement of CNV lesion area in both experimentally-induced and pathological CNV. The results indicate that MOT could be a useful research tool in helping evaluate the effects of therapeutics on CNV growth.

In Vivo Efficacy of an Injectable Microsphere-Hydrogel Ocular Drug Delivery System

ABSTRACT Purpose: Demonstrate in vivo that controlled and extended release of a low dose of anti-vascular endothelial growth factor (anti-VEGF) from a microsphere-hydrogel drug delivery system (DDS) has a therapeutic effect in a laser-induced rat model of choroidal neovascularization (CNV). Methods: Anti-VEGF (ranibizumab or aflibercept) was loaded into poly(lactic-co-glycolic acid) microspheres that were then suspended within an injectable poly(N-isopropylacrylamide)-based thermo-responsive hydrogel DDS.The DDS was shown previously to release bioactive anti-VEGF for ~200 days. CNV was induced using an Ar-green laser. The four experimental groups were as follows: (i) non-treated, (ii) drug-free DDS, (iii) anti-VEGF-loaded DDS, and (iv) bolus injection of anti-VEGF. CNV lesion areas were measured based on fluorescein angiograms and quantified using a multi-Otsu thresholding technique. Intraocular pressure (IOP) and dark-adapted electroretinogram (ERG) were also obtained pre- and post-treatment (1, 2, 4, 8, and 12 weeks). Results: The anti-VEGF-loaded DDS group had significantly smaller (60%) CNV lesion areas than non-treated animals throughout the study. A small transient increase in IOP was seen immediately after injection; however, all IOP measurements at all time points were within the normal range. There were no significant changes in ERG maximal response compared to pre-treatment measurements for the drug-loaded DDS, which suggests no adverse effects on retinal cellular function. Conclusions: The current study demonstrates that the DDS can effectively decrease laser-induced CNV lesions in a murine model. Controlled and extended release from our DDS achieved greater treatment efficacy using an order of magnitude less drug than what is required with bolus administration. This suggests that our DDS may provide a significant advantage in the treatment of posterior segment eye diseases.

Dual Electroretinogram/Nitric Oxide Carbon Fiber Microelectrode for Direct Measurement of Nitric Oxide in the In Vivo Retina

Nitric oxide (NO) plays an important physiological role in normal and pathological retinas. Intraretinal NO concentrations have not been directly measured due to lack of NO electrodes capable of determining their location in the retina. The microelectrodes described here allow recording of the intraretinal electroretinogram (ERG) and NO concentration from the same location, with ERGs used to determine retinal depth. Double-barreled electrodes were constructed with one barrel serving as a reference/voltage recording barrel and the other containing a Nafion-coated carbon fiber used to detect NO amperometrically. Nafion coating imparted a high selectivity for NO versus ascorbic acid (2000:1). In vivo rodent experiments demonstrated that the electrodes could record intraretinal ERGs and NO current with minimal retinal thickness deformation (9%), allowing for retinal NO depth profile measurements. Comparison of NO depth profiles under control conditions and under nitric oxide synthase (NOS) inhibition by 5 mM L-NG-Nitroarginine methyl ester (L-NAME) verified that the recorded current was attributable to NO. NO concentrations from control profiles ( n = 4) were 2.37 ± 0.34 μM at the choroid and 1.12 ± 0.14 μM at the retinal surface. NO concentrations from L-NAME profiles ( n = 4) were significantly lower at 0.83 ± 0.15 μM at the choroid ( p = 0.006) and 0.27 ± 0.04 μM at the retinal surface ( p = 0.001). Localized regions of increased NO (100-400 nM) were seen in the inner retina under control conditions but not after L-NAME. The dual ERG-NO electrode may be a valuable tool in evaluating the role of NO in normal and diseased retinas.

Design of a Novel Electronic Travel Aid to Assist Visually Impaired Individuals Navigate Their Environment

Worldwide there are over 160 million people with severe visual impairment, as defined by visual acuity poorer than 20/200.1 A prominent concern for visually impaired individuals is their limited navigational abilities due to insufficient sensory information about their surrounding environment which results in difficulty with navigating new or complex environments. In these situations, they often have to rely on the assistance of others to help them reach their destination. Furthermore, even when the visually impaired individuals are familiar with the area, they are not always aware of non-stationary obstacles, such as cars or people. Two commonly used solutions currently available to help visually impaired individuals navigate their surroundings are the white cane and guide dogs. The white cane is useful for alerting its users to obstacles closer than 1.5 m, but it does not provide any information about the environment beyond that scope. Guide dogs are in unfortunately limited supply and can cost upwards of

Biodegradable Microsphere-Hydrogel Ocular Drug Delivery System for Controlled and Extended Release of Bioactive Aflibercept In Vitro

42,000 to train.2 To address this challenge, multiple groups have examined more technologically advanced solutions to help visually impaired individuals. However, these devices have some major limitations, such as complicated display modalities and non-intuitive sensory representation of environmental information. The major goal of this proposal is to develop a new electronic travel aid (ETA) that will help visually impaired individuals navigate their environment more easily by using a novel method of directly displaying the location of obstacles up to 4 m away on the user’s torso with a grid of small vibrational devices called tactors. This device is intended to be used with a traditional white cane that can detect objects very close to the user and terrain changes, such as a step in a stairwell.Copyright