pH Sensitive Erythrocyte-Derived Membrane for Acute Systemic Retention and Increased Infectivity of Coated Oncolytic Vaccinia Virus.

Oncolytic viruses have emerged as a promising modality in cancer treatment given their high synergy with highly efficient immune checkpoint inhibitors. However, their potency is limited by their rapid in vivo clearance. To overcome this, we coated oncolytic vaccinia viruses (oVV) with erythrocyte-derived membranes (EDMs), hypothesizing that they would not only remain in systemic circulation for longer as erythrocytes would when administered intravenously, but also respond to environmental pH cues due to their membrane surface sialic acid residues. For this, we developed a model based on DLVO theory to show that the acidic moieties on the surface of EDM confers it the ability to respond to pH-based stimuli. We corroborate our modeling results through in vitro cell culture models and show that EDM-coated oVV infects cancer cells faster under acidic conditions akin to the tumor microenvironment. When EDM-coated oVVs were intravenously injected into wild-type mice, they exhibited prolonged circulation at higher concentrations when compared to the unprocessed oVV. Furthermore, when EDM-coated oVV was directly injected into xenografted tumors, we observed that they were suppressed earlier than the tumors that received regular oVV, suggesting that the EDM coating does not hinder oVV infectivity. Overall, we found that EDM was able to serve as a multi-functional encapsulant that allowed the payload to remain in circulation at higher concentrations when administered intravenously while simultaneously exhibiting pH-responsive properties.

Ultrasound-Enhanced Transcorneal Drug Delivery for Treatment of Fungal Keratitis.

PURPOSE: Transcorneal drug delivery is hindered by ocular physical and biochemical properties, such as tear production, the epithelial layer of the cornea, and blinking. The aim of this study was to determine whether ultrasound can be applied to increase the transcorneal drug delivery of natamycin used in the treatment of fungal keratitis without dangerously overheating the surrounding ocular tissues. METHODS: To verify the safety of various sets of ultrasound parameters, modeling studies were conducted using OnScale, an ultrasonic wave modeling software. Ultrasound parameters determined optimal for ocular tissue safety were used in a laboratory setting in a jacketed Franz diffusion cell setup. Histological images of the cross-section of the corneas used in experiments were examined for cell damage under a microscope. RESULTS: Increases in transcorneal drug delivery were seen in every treatment parameter combination when compared with the sham treatment. The highest increase was 4.0 times for 5 minutes of pulsed ultrasound at a 25% duty cycle and a frequency of 400 kHz and an intensity of 0.5 W/cm 2 with statistical significance ( P < 0.001). Histological analysis revealed structural damage only in the corneal epithelium, with most damage being at the epithelial surface. CONCLUSIONS: This study suggests that ultrasound is a safe, effective, and minimally invasive treatment method for enhancing the transcorneal drug delivery of natamycin. Further research is needed into the long-term effects of ultrasound parameters used in this study on human ocular tissues.

Therapeutic Ultrasound-Enhanced Transcorneal PHMB Delivery In Vitro.

OBJECTIVE: Delivery of therapeutic agents to the cornea is a difficult task in the treatment of parasitic keratitis. In this study, we looked at using different combinations of ultrasound parameters to enhance corneal permeability to polyhexamethylene biguanide (PHMB), a clinically available ophthalmic antiparasitic formulation. METHODS: Permeability of PHMB was investigated in vitro using a standard diffusion cell setup. Continuous or 25% duty-cycle ultrasound was used at frequencies of 400 or 600 kHz, intensities of 0.5 or 0.8 W/cm2 , and exposure times ranging from 1 to 5 minutes. Structural changes in the cornea were examined using light microscopy. RESULTS: Ultrasound exposure produced increases in transcorneal delivery in every treatment parameter combination when compared to the sham treatment. The highest increase was 2.36 times for 5 minutes of continuous ultrasound at a frequency of 600 kHz and an intensity of 0.5 W/cm2 with statistical significance (p <.001). Histological analysis showed that ultrasound application only caused structural changes in the corneal epithelium, with most damage being at the surface layers. CONCLUSIONS: This study suggests the possibility of therapeutic ultrasound as a novel drug delivery technique for the treatment of parasitic keratitis. Further studies are needed to examine the thermal effects of these proposed ultrasound applications and the long-term viability of this treatment.

Ultrasound-Enhanced Drug Delivery for Treatment of Acanthamoeba Keratitis.

Reaching sufficient amounts of therapeutic agents in ocular tissues is a major challenge in ophthalmology. In this study, we examined the effects of ultrasound application for delivery of polyhexamethylene biguanide for treatment of Acanthamoeba keratitis. Ultrasound intensities of 0.5 - 0.8 W/cm2 and frequencies of 400 - 600 kHz were tested with exposure durations of 1 - 5 minutes. Light microscopy was used to determine the ultrasound-induced structural changes in the cornea. All groups showed increases in drug concentration, up to 2.36 times, passing through the cornea, with the 600 kHz treatment groups reaching statistical significance. Structural changes were observed in the epithelial layer of the cornea, but the stroma and endothelium remained mostly unaffected.