TY - JOUR
T1 - Enzymatic degradation identifies components responsible for the structural properties of the vitreous body
AU - Filas, Benjamen A.
AU - Zhang, Qianru
AU - Okamoto, Ruth J.
AU - Shui, Ying Bo
AU - Beebe, David C.
PY - 2014/1/3
Y1 - 2014/1/3
N2 - PURPOSE. Vitreous degeneration contributes to several age-related eye diseases, including retinal detachment, macular hole, macular traction syndrome, and nuclear cataracts. Remarkably little is understood about the molecular interactions responsible for maintaining vitreous structure. The purpose of this study was to measure the structural properties of the vitreous body after enzymatic degradation of selected macromolecules. METHODS. Mechanical properties of plugs of bovine and porcine vitreous were analyzed using a rheometer. Oscillatory and extensional tests measured vitreous stiffness and adhesivity, respectively. Major structural components of the vitreous were degraded by incubation overnight in collagenase, trypsin, or hyaluronidase, singly or in combination. Vitreous bodies were also incubated in hyper- or hypotonic saline. Effects of these treatments on the mechanical properties of the vitreous were measured by rheometry. RESULTS. Enzymatic digestion of each class of macromolecules decreased the stiffness of bovine vitreous by approximately half (P < 0.05). Differential effects were observed on the damping capacity of the vitreous (P < 0.05), which was shown to correlate with material behavior in extension (P < 0.01). Digestion of hyaluronan significantly decreased the damping capacity of the vitreous and increased adhesivity. Collagen degradation resulted in the opposite effect, whereas digestion of proteins and proteoglycans with trypsin did not alter behavior relative to controls. Osmotic perturbations and double-enzyme treatments further implicated hyaluronan and hyaluronan-associated water as a primary regulator of adhesivity and material behavior in extension. CONCLUSIONS. Collagen, hyaluronan, and proteoglycans act synergistically to maintain vitreous stiffness. Hyaluronan is a key mediator of vitreous adhesivity, and mechanical damping is an important factor influencing dynamic vitreous behavior.
AB - PURPOSE. Vitreous degeneration contributes to several age-related eye diseases, including retinal detachment, macular hole, macular traction syndrome, and nuclear cataracts. Remarkably little is understood about the molecular interactions responsible for maintaining vitreous structure. The purpose of this study was to measure the structural properties of the vitreous body after enzymatic degradation of selected macromolecules. METHODS. Mechanical properties of plugs of bovine and porcine vitreous were analyzed using a rheometer. Oscillatory and extensional tests measured vitreous stiffness and adhesivity, respectively. Major structural components of the vitreous were degraded by incubation overnight in collagenase, trypsin, or hyaluronidase, singly or in combination. Vitreous bodies were also incubated in hyper- or hypotonic saline. Effects of these treatments on the mechanical properties of the vitreous were measured by rheometry. RESULTS. Enzymatic digestion of each class of macromolecules decreased the stiffness of bovine vitreous by approximately half (P < 0.05). Differential effects were observed on the damping capacity of the vitreous (P < 0.05), which was shown to correlate with material behavior in extension (P < 0.01). Digestion of hyaluronan significantly decreased the damping capacity of the vitreous and increased adhesivity. Collagen degradation resulted in the opposite effect, whereas digestion of proteins and proteoglycans with trypsin did not alter behavior relative to controls. Osmotic perturbations and double-enzyme treatments further implicated hyaluronan and hyaluronan-associated water as a primary regulator of adhesivity and material behavior in extension. CONCLUSIONS. Collagen, hyaluronan, and proteoglycans act synergistically to maintain vitreous stiffness. Hyaluronan is a key mediator of vitreous adhesivity, and mechanical damping is an important factor influencing dynamic vitreous behavior.
KW - Dynamic mechanical analysis
KW - Enzymatic vitreolysis
KW - Liquefaction
KW - Mechanical properties
KW - Ocular biomechanics
KW - Vitreous
KW - Vitreous degeneration
UR - http://www.scopus.com/inward/record.url?scp=84891642378&partnerID=8YFLogxK
U2 - 10.1167/iovs.13-13026
DO - 10.1167/iovs.13-13026
M3 - Article
AN - SCOPUS:84891642378
SN - 0146-0404
VL - 55
SP - 55
EP - 63
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
IS - 1
ER -