TY - JOUR
T1 - Determining vitreous viscosity using fluorescence recovery after photobleaching
AU - Srikantha, Nishanthan
AU - Teijeiro-Gonzalez, Yurema
AU - Simpson, Andrew
AU - Elsaid, Naba
AU - Somavarapu, Satyanarayana
AU - Suhling, Klaus
AU - Jackson, Timothy L.
N1 - Funding Information:
Funding: The project was funded by Fight for Sight Grant Number 1874 and BBSRC Grant Number BB/ R004803/1. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. N. Srikantha is a Fight for Sight Research Fellow. K. Suhling acknowledges BBSRC grant BB/R004803/ 1.
Publisher Copyright:
© 2022 Public Library of Science. All rights reserved.
PY - 2022/2
Y1 - 2022/2
N2 - PURPOSE: Vitreous humor is a complex biofluid whose composition determines its structure and function. Vitreous viscosity will affect the delivery, distribution, and half-life of intraocular drugs, and key physiological molecules. The central pig vitreous is thought to closely match human vitreous viscosity. Diffusion is inversely related to viscosity, and diffusion is of fundamental importance for all biochemical reactions. Fluorescence Recovery After Photobleaching (FRAP) may provide a novel means of measuring intravitreal diffusion that could be applied to drugs and physiological macromolecules. It would also provide information about vitreous viscosity, which is relevant to drug elimination, and delivery.METHODS: Vitreous viscosity and intravitreal macromolecular diffusion of fluorescently labelled macromolecules were investigated in porcine eyes using fluorescence recovery after photobleaching (FRAP). Fluorescein isothiocyanate conjugated (FITC) dextrans and ficolls of varying molecular weights (MWs), and FITC-bovine serum albumin (BSA) were employed using FRAP bleach areas of different diameters.RESULTS: The mean (±standard deviation) viscosity of porcine vitreous using dextran, ficoll and BSA were 3.54 ± 1.40, 2.86 ± 1.13 and 4.54 ± 0.13 cP respectively, with an average of 3.65 ± 0.60 cP.CONCLUSIONS: FRAP is a feasible and practical optical method to quantify the diffusion of macromolecules through vitreous.
AB - PURPOSE: Vitreous humor is a complex biofluid whose composition determines its structure and function. Vitreous viscosity will affect the delivery, distribution, and half-life of intraocular drugs, and key physiological molecules. The central pig vitreous is thought to closely match human vitreous viscosity. Diffusion is inversely related to viscosity, and diffusion is of fundamental importance for all biochemical reactions. Fluorescence Recovery After Photobleaching (FRAP) may provide a novel means of measuring intravitreal diffusion that could be applied to drugs and physiological macromolecules. It would also provide information about vitreous viscosity, which is relevant to drug elimination, and delivery.METHODS: Vitreous viscosity and intravitreal macromolecular diffusion of fluorescently labelled macromolecules were investigated in porcine eyes using fluorescence recovery after photobleaching (FRAP). Fluorescein isothiocyanate conjugated (FITC) dextrans and ficolls of varying molecular weights (MWs), and FITC-bovine serum albumin (BSA) were employed using FRAP bleach areas of different diameters.RESULTS: The mean (±standard deviation) viscosity of porcine vitreous using dextran, ficoll and BSA were 3.54 ± 1.40, 2.86 ± 1.13 and 4.54 ± 0.13 cP respectively, with an average of 3.65 ± 0.60 cP.CONCLUSIONS: FRAP is a feasible and practical optical method to quantify the diffusion of macromolecules through vitreous.
UR - http://www.scopus.com/inward/record.url?scp=85124412693&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0261925
DO - 10.1371/journal.pone.0261925
M3 - Article
C2 - 35143514
SN - 1932-6203
VL - 17
JO - PLoS ONE
JF - PLoS ONE
IS - 2
M1 - e0261925
ER -