Multikinase inhibition-mediated proliferative vitreoretinopathy therapy by nanoparticles in rabbits

Abstract

Purpose: To investigate the efficacy of nanoparticles in treating proliferative vitreoretinopathy (PVR) through clinical observation, histology, and immunohistochemistry, despite unsatisfactory surgical outcomes and failed therapies for the current PVR treatment.

Design: Twelve rabbits were divided into control and nintedanib (NTB) groups. The rabbits underwent weekly ophthalmologic examinations over a period of four weeks.

Methods: At the end of the fourth week, the rabbits' eyes were removed for histological and immunohistochemical evaluation. Three additional rabbits outside the PVR model were administered a 0.5% NTB-loaded liposomal formulation in one eye. The drug concentrations in the vitreous samples were determined using high-pressure liquid chromatography on days 1, 7, 14, and 35.

Results: The PVR stages were low in the NTB group, and there was no significant difference between the NTB and control groups (p = 0.108). However, it is worth noting that the group treated with NTB had significantly fewer epiretinal membrane formations during the histological evaluation. In addition, the corrected fluorescence intensity measurement of the subjects for collagen-1 in the NTB group was significantly lower than that in the control group (p = 0.004). Most importantly, no significant adverse effects were observed.

Conclusions: Our study has provided preclinical support for a liposomal formulation containing NTB that, with single-dose administration, has the potential to be effective in vivo in preventing the development of PVR and its correlated pathologies without causing any significant side effects.

Figure 1

(A) Merangiotic structure of rabbit fundus and optic disc. (B) Vitreous hemorrhage observed immediately after application.

Figure 2

Size dimension and zeta potential of the nintedanib-liposome (NTB-LIP) formulations.

Figure 3

Microscope images of the NTB-LIP formulations.

Figure 4

High-pressure liquid chromatograms before and after liposome addition.

Figure 5

Expression levels of specific markers, whether treated with formulations or not. A: Formulations’ marker levels. B: Fluorescence microscope image of the RPE cells after staining. C: Western blot protein dragging result (Nf-ĸβ: necrotic factor kappa beta; TNF-ɑ: tumor necrosis factor alpha; VEGF-A: vascular endothelial growth factor A).

Figure 6

Viability assay to evaluate the formulations’ toxicological formations.

Figure 7

Migration study microscope images.

Figure 8

Dissolution profile of the NTB and NTB-LIP formulations.

Figure 9

Fundus images of the rabbits. A: Total retinal detachment; the optic disc is indicated by an arrow (↑;subject no. C1, stage 5). B: Focal traction and intravitreal membrane formations and gliotic changes in the retina (subject no. C2, stage 2). C: Traction creating a membrane (↑) and a wide detachment area (De; subject no. C4, stage 4). D: Intravitreal membrane, vitreous opacities, and gliotic changes in the retina (subject no. N1, stage 1). E: Membrane extending from the disc (OD) and that does not produce traction, indicated by an arrow (↑;subject no. N2, stage 1). (F) Intravitreal membrane indicated by an arrow (↑), and gliotic changes in the retina (subject no. N6, stage 1).

Figure 10

Vascular tissue (←) attached to the lens (100×).

Figure 11

A choriocapillary layer (star) thinner in the NTB group (left) than in the control group (right; 40×).

Figure 12

Fibrotic membrane formations in the NTB (left) and control (right) groups. The asterisk indicates fibrotic membranes (40×).

Figure 13

Histology sections of the subjects in the NTB group. A: Papillary changes (*; 40×). B: Corrugation in the retina (star; 100×). C: Opaque region (↑; 12.5×).

Figure 14

Corrected fluorescence intensity measurement (CFIM) results of the NTB and control groups. Statistical analysis (Mann–Whitney U test) indicated that the CFIM level of the NTB group was significantly lower than that of the control group (p = 0.004).