Nanoparticle-hydrogel composite as dual-drug delivery system for the potential application of corneal graft rejection.

Immune rejection remains the major cause of corneal graft failure. Immunosuppressants (such as rapamycin; RAPA) adjunctive to antibiotics (such as levofloxacin hydrochloride; Lev) are a clinical mainstay after corneal grafts but suffer from poor ocular bioavailability associated with severe side effects. In this study, we fabricated a Lev@RAPA micelle loaded cationic peptide-based hydrogel (NapFFKK) as a dual-drug delivery system by integrating RAPA micelles with Lev into a cationic NapFFKK hydrogel to potentially reduced the risk of corneal graft rejection. The properties of the resulting hydrogels were characterized using transmission electronmicroscopy and rheometer. Lev@RAPA micelles loaded NapFFKK hydrogel provided sustained in vitro drug release without compromising their inherent pharmacological activities. Topical instillation of Lev@RAPA micelles loaded NapFFKK hydrogel resulted in the great ocular tolerance and extended precorneal retention over 60 min, thus significantly enhancing the ocular bioavailability of both Lev and RAPA. Overall, such dual-drug delivery system might be a promising formulation for the suppression of corneal graft failure.

Subconjunctival injection of rapamycin-loaded polymeric microparticles for effective suppression of noninfectious uveitis in rats.

Noninfective uveitis is a major cause of vision impairment, and corticosteroid medication is a mainstay clinical strategy that causes severe side effects. Rapamycin (RAPA), a potent immunomodulator, is a promising treatment for noninfective uveitis. However, because high and frequent dosages are required, it is a great challenge to implement its clinical translation for noninfective uveitis therapy owing to its serious toxicity. In the present study, we engineered an injectable microparticulate drug delivery system based on biodegradable block polymers (i.e., polycaprolactone-poly (ethylene glycol)-polycaprolactone, PCEC) for efficient ocular delivery of RAPA via a subconjunctival injection route and investigated its therapeutic efficacy in an experimental autoimmune uveitis (EAU) rat model. RAPA-PCEC microparticles were fabricated using the emulsion-evaporation method and thoroughly characterized using scanning electron microscopy, fourier transform infrared spectroscopy, X-ray diffraction, and differential scanning calorimetry. The formed microparticles exhibited slow in vitro degradation over 28 days, and provided both in vitro and in vivo sustained release of RAPA over 4 weeks. Additionally, a single subconjunctival injection of PCEC microparticles resulted in high ocular tolerance. More importantly, subconjunctival injection of RAPA-PCEC microparticles significantly attenuated the clinical signs of EAU in a dose-dependent manner by reducing inflammatory cell infiltration (i.e., CD45+ cells and Th17 cells) and inhibiting microglial activation. Overall, this injectable microparticulate system may be promising vehicle for intraocular delivery of RAPA for the treatment of noninfective uveitis.

Repeatability and agreement of AOCT-1000 M, RTVue XR and IOL master 500 in measuring corneal thickness mapping and axial length applying principle of optical coherence tomography.

PURPOSE: To evaluate the repeatability and agreement of Fourier-domain optical coherence tomography (AOCT-1000 M and RTVue XR) and partial coherence interferometry biometer (IOL Master 500) in measuring corneal thickness mapping and axial length respectively. METHODS: Corneal thickness was measured by AOCT-1000 M and RTVue XR. Axial lengths were measured by AOCT-1000 M and IOL Master 500. The repeatability and agreement of corneal thickness and axial length were calculated in two groups of devices. The intraclass correlation coefficient (ICC) was used to verify the repeatability of the device. The 95% confidence interval of the difference compared to the set cut-off value was used to verify the agreement between the two devices. RESULTS: A total of 60 subjects with 58 eyes were included. The central corneal thickness measured by AOCT-1000 M and RTVue XR were 504.46 ± 42.53 µm and 504.43 ± 42.89 µm respectively. The average difference between groups was 0.03 ± 4.58 µm, and the 95% confidence interval was (-1.17, 1.24), which was far less than the set threshold value of 15 µm (P < 0.001). Both RTVue XR and AOCT-1000 M had very good ICC values of central corneal thickness (0.998 and 0.994, respectively). The average axial lengths measured by AOCT-1000 M and IOL Master 500 were 24.28 ± 1.25 mm and 24.29 ± 1.26 mm respectively and the 95% confidence interval was (-0.02, 0.01), which was less than the set threshold value of 0.15 mm (P < 0.001). The ICC for both devices were 1.000. CONCLUSION: Good repeatability and agreement were seen in measurements of central corneal thickness and axial length by AOCT-1000 M.

Changes in Shape Discrimination Sensitivity Under Glare Conditions After Orthokeratology in Myopic Children: A Prospective Study.

Purpose: To investigate changes in shape discrimination under mesopic conditions with and without glare after orthokeratology in myopic children. Methods: This prospective study included 79 eyes of 79 myopic children (ages: 8-16 years). Shape discrimination thresholds (SDTs) were measured using radial frequency patterns, with a radial frequency of 4 cycles/360°, a peak spatial frequency of 3 cycles per degree, a contrast of 20%, and a mean radius of 1.5 degrees. SDT under mesopic conditions with and without glare was measured before orthokeratology and again at 1 week and 1 month after orthokeratology. Changes in the SDTs and their relationships to baseline ocular parameters were analyzed. Results: SDTs with glare decreased significantly at 1 week (-0.08 ± 0.15 log(arcsec), P < 0.001) and 1 month (-0.09 ± 0.15 log(arcsec), P < 0.001) after orthokeratology. SDTs without glare remained stable (P = 0.81 and P = 1.00, respectively). The difference between SDTs with and without glare also decreased significantly at 1 week (-0.10 ± 0.17 log(arcsec), P < 0.001) and at 1 month (-0.08 ± 0.18 log(arcsec), P = 0.001) after orthokeratology. Based on a multivariate analysis, the greater decrease in SDT with glare after 1 month of orthokeratology was associated with a higher baseline spherical equivalent refraction. Conclusions: Orthokeratology resulted in improved shape discrimination in myopic children under mesopic conditions but only when measured in the presence of glare.

An injectable thermosensitive hydrogel for dual delivery of diclofenac and Avastin® to effectively suppress inflammatory corneal neovascularization.

Corneal neovascularization (CNV) is a sequela of anterior segment inflammation, which could lead to vision impairment and even blindness. In the present study, the dual delivery of anti-inflammatory agent (i.e., diclofenac; DIC) and anti-VEGF antibody (i.e., Avastin®; Ava) by the thermosensitive hydrogel (Poly(dl-lactide)-poly(ethylene glycol)-poly(dl-lactide); PDLLA-PEG-PDLLA) is expected to effectively inhibit CNV via their synergistic effects. The optimal DIC micelles were formulated and then mixed with Ava and PDLLA-PEG-PDLLA aqueous solution to generate various DIC@Ava-loaded hydrogels. The co-encapsulation of DIC micelles and Ava did not influence the gelling behavior of the system, and the resulting DIC@Ava-loaded hydrogel provided sustained drug release of both DIC and Ava without compromising their pharmacological activity over 19 days. As indicated by in vitro cytotoxicity and in vivo ocular biocompatibility test, the proposed PDLLA-PEG-PDLLA hydrogel caused minimal cytotoxicity against all tested cell lines at a polymeric concentration ranging from 0.05 mg/mL to 0.8 mg/mL and demonstrated good ocular biocompatibility after a single subconjunctival injection. Using the rabbit CNV model, we documented the superior anti-angiogenic effects of the DIC@Ava-loaded hydrogel over Ava alone medication (treatment with Ava solution and Ava-loaded hydrogel) due to synergistic effects of anti-VEGF and anti-inflammatory action. Overall, the proposed DIC@Ava-loaded hydrogel might be a powerful strategy to reduce CNV.

Biofunctional peptide-click PEG-based hydrogels as 3D cell scaffolds for corneal epithelial regeneration.

Poly(ethylene glycol) (PEG)-based hydrogels as highly promising 3D cell scaffolds have been widely implemented in the field of tissue regrowth and regeneration, yet the functionalized PEG hydrogel providing dynamic, cell-instructive microenvironments is inherently difficult to obtain. Here, we have exploited the specificity of click reaction to develop a set of hydrogels based on 4-arm PEG tetraazide (4-arm-PEG-N3) and di-propargylated peptides (GRGDG and GRDGG) with tunable physicochemical properties applicable for 3D cell scaffolds. The azide groups of PEG were reacted with the alkynyl groups of peptides, catalyzed by copper to form triazole rings, thus generating a cross-linked hydrogel. The gelation time and mechanical strength of the hydrogels varied according to the PEG/peptide feed ratio. The resulting hydrogel exhibited a typical porous microstructure and suitable swelling behavior. The in vitro cytotoxicity test indicated that the resulting hydrogels did not cause apparent cytotoxicity against human corneal epithelial cells (HCECs). After co-incubation with HCECs, the density of RGD as well as peptide sequence in the hydrogels remarkably affected the cell attachment, spreadability, and proliferation. Additionally, the proposed hydrogel showed high ocular biocompatibility after being embedded subconjunctivally into rabbit eyes. Overall, these findings highlighted that the biofunctional hydrogels formed by PEG and RGD motifs via a controllable click reaction might be promising 3D cell scaffolds for corneal epithelial regeneration.

Single subcutaneous injection of the minocycline nanocomposite-loaded thermosensitive hydrogel for the effective attenuation of experimental autoimmune uveitis.

Autoimmune uveitis induces a serious pathological and inflammatory response in the retina/choroid and results in vision impairment and blindness. Here, we report a minocycline (Mino) nanocomposite-loaded hydrogel offering a high drug payload and sustained drug release for the effective control of ocular inflammation via a single subcutaneous injection. In the presence of divalent cations (i.e., Ca2+), Mino was found to co-assemble with a phosphorylated peptide (i.e., NapGFFpY) via electrostatic interaction and consequently generating Mino nanocomposite. The drug entrapment efficiency (EE) of the Mino nanocomposite varied from 29.93 ± 0.76% to 67.90 ± 6.57%, depending on different component concentrations. After incorporation into 30 wt% poly (D,L-lactide)-b-poly (ethylene glycol)-b-poly (D,L-lactide) (PDLLA-PEG-PDLLA) thermosensitive hydrogel, the resulting Mino nanocomposite-loaded hydrogel provided a sustained drug release over 21 days. In the experimental autoimmune uveitis (EAU) rat model, a single subcutaneous injection of the Mino nanocomposite-loaded hydrogel effectively alleviated ocular inflammation in a dose-dependent manner. As indicated by optical coherence tomography (OCT) and electroretinogram (ERG) measurements, the Mino nanocomposite-loaded hydrogel treatment not only remarkably reduced destruction of the retina by EAU, but also greatly rescued retinal functions. Moreover, the proposed Mino nanocomposite-loaded hydrogel exerted its therapeutic effect on EAU primarily through a significant reduction of the influx of leukocytes and Th17 cells as well as suppression of microglia activation and apoptosis in the retina. Overall, the proposed Mino nanocomposite-loaded hydrogel might be a promising strategy for the clinical management of EAU.

Evaluation of microvasculature alterations in convalescent Vogt-Koyanagi-Harada disease using optical coherence tomography angiography.

PURPOSE: To evaluate the microvasculature alterations in convalescent Vogt-Koyanagi-Harada (VKH) disease using optical coherence tomography angiography (OCTA), and to explore the association between microvasculature and the presence of sunset glow fundus (SGF). METHODS: A cross-sectional study was conducted with 28 VKH patients at convalescent stage and 25 healthy individuals. Both eyes of each participant were enrolled. The VKH patients were classified into two subgroups based on the existence of SGF. OCTA images (3 × 3 mm) were assessed for the data of superficial capillaris plexus (SCP), deep capillaris plexus (DCP), choriocapillaris, and foveal avascular zone (FAZ). RESULTS: Compared with healthy control eyes and eyes without SGF, the vessel densities of the SCP and DCP decreased significantly in most regions of eyes with SGF (p < 0.0167). No significant difference of vascular perfusion was found between eyes without SGF and control eyes (p > 0.05). VKH patients with SGF had slightly increased FAZ area (p = 0.067) and decreased choroid flow area (p = 0.427) than those in the control group. CONCLUSION: Convalescent VKH patients with SGF showed decreased macular capillary perfusion. OCTA could serve as a sensitive tool to assess the microvasculature alterations of VKH disease.

Glycol chitosan/oxidized hyaluronic acid hydrogel film for topical ocular delivery of dexamethasone and levofloxacin.

In the present study, we fabricated a glycol chitosan/oxidized hyaluronic acid hydrogel film with promising potential for the dual ophthalmic delivery of dexamethasone (Dex) and levofloxacin (Lev). Utilizing different oxidation degrees of oxidized hyaluronic acid (OHA), several blank hydrogel films and Lev-loaded hydrogel films were successfully fabricated. With an increase in the oxidation degree of OHA, the swelling ratio of the hydrogel films decreased accordingly. The hydrogel films displayed a stepwise release of Lev and Dex, with Lev rapidly released from the hydrogel film, followed by a sustained release of Dex. Lev-loaded hydrogel films revealed a potent capacity to inhibit bacterial growth in different bacterial strains. In lipopolysaccharide-activated RAW264.7 macrophages, the formulated hydrogel films displayed potent in vitro anti-inflammatory activity by significantly downregulating various inflammatory cytokines. Overall, the fabricated hydrogel film acting as a dual drug delivery system might be a promising vehicle for the treatment of postoperative endophthalmitis.

Mucoadhesive dexamethasone-glycol chitosan nanoparticles for ophthalmic drug delivery.

Self-assembly of drug-polysacrrides conjugates forming nanostructures provides a simple and promising strategy for the extension of precorneal retention and enhancement of corneal permeability. In the present study, a series of dexamethasone-glycol chitosan (Dex-GCS) conjugates were synthesized and thoroughly characterized by Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD) and ultraviolet-visible (UV-Vis) spectroscopy. The resulting Dex-GCS conjugates were able to self-assemble into nanoparticles spontaneously with particle sizes in the range of 277-289 nm and a positive charge of approximately +15 mV. Roughly spherical nanoparticles were observed by transmission electron microscopy (TEM). The in vitro mucoadhesive properties of Dex-GCS nanoparticles were evaluated by recording the variations in the zeta potential after incubation with different concentrations of mucin. In vitro release studies performed in phosphate-buffered saline (PBS, pH = 7.4) indicated progressive Dex release up to 8 h, followed by a plateau up to 48 h. Dex-GCS nanoparticles caused slight cytotoxicity against L929, HCEC and RAW 264.7 cells after 24 h of incubation and displayed a nearly identical anti-inflammatory efficacy to dexamethasone sodium phosphate (Dexp) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. More importantly, the proposed Dex-GCS nanoparticles showed good ocular tolerance and provided a relatively longer precorneal duration compared with that of the aqueous solution formulation, which suggested that the self-assembled Dex-GCS nanoparticle might be a promising candidate for ophthalmic drug delivery.

Thermosensitive glycol chitosan-based hydrogel as a topical ocular drug delivery system for enhanced ocular bioavailability.

In the present study, we developed and evaluated an in situ gelling system based on hexanoyl glycol chitosan (H-GCS) for enhanced ocular bioavailability. An aqueous solution of H-GCS exhibited a typical sol-gel transition at 32 °C. The formed H-GCS hydrogel was characterized by rheology and scanning electron microscopy (SEM). H-GCS had minimal in vitro cytotoxicity against L-929 and HCEC cells over a concentration range of 0-0.8 mg/mL. Additionally, the H-GCS hydrogel exhibited good ocular tolerance and biocompatibility after a single instillation. Moreover, H-GCS hydrogel significantly prolonged the precorneal retention of fluorescein sodium compared with its aqueous solution. An in vivo pharmacokinetic study demonstrated that the levofloxacin-loaded H-GCS hydrogel could provide a significantly higher Cmax and AUC0-12h compared with the levofloxacin aqueous solution, thus increasing ocular bioavailability. Overall, the proposed H-GCS hydrogel acts as an in situ gelling system that might represent a promising vehicle for topical ocular drug delivery.

A Facile Strategy to Generate High Drug Payload Celecoxib Micelles for Enhanced Corneal Permeability.

Topical ocular drug administration aimed at providing a high drug concentration at the precorneal site accompanied with enhanced corneal permeability to avoid systemic side effects is a very important therapeutic goal in ocular disorder therapy. In the present study, the solubility of the poorly soluble drug celecoxib (CXB) was significantly improved using a facile strategy to generate a high drug payload micellar formulation. By varying the drug/polymer feed ratios, the mean diameter of the formed CXB micelles ranged from 21.34 ± 0.23 to 28.53 ± 0.11 nm, and the drug loading capacity ranged from 4.31 to 15.87%. Transmission electron microscopy (TEM) showed that the formed CXB micelles had a uniform spherical morphology. Differential scanning calorimetry (DSC), X-ray powder diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) were used to characterize the physicochemical properties of lyophilized CXB micelles. The obtained CXB micelles retained their properties through freeze-drying and rehydration, thereby providing long-term physicochemical stability over 3 months of storage at -20 °C. An in vitro release study showed that the CXB micelles released CXB in a sustained release manner without any apparent burst release over 72 h by the Higuchi non-Fickian diffusion mechanism. Notably, using corneas excised from rabbits, the in vitro corneal permeability of CXB from the micellar formulation was observed to be significantly greater than that of the microparticle formulation. Overall, the proposed micelles might be a promising vehicle for ophthalmic delivery of CXB with the significant enhancement of water solubility and corneal permeability.