ocular drugs influence on the structure of ocular tissues

Saratov State University

______________________________________________

Department of Optics & Biophotonics

__________________________________________________

Saratov Fall Meeting 2011

Ocular drugs influence on the structure of ocular tissues

Anzhelika S. Orlova, Tatyana G. Kamenskikh, Ivan D. Kamenskikh,

Igor O. Kolbenev,

Saratov State Medical University

Alexey N. Bashkatov, Elina A. Genina, Artem A. Aratov,Anastasiya M. Parkheychuk, Valery V. Tuchin

Department of Optics and Biophotonics of Saratov State University

e-mail: [email protected]

Anzhelika S. Orlova, Tatyana G. Kamenskikh, Ivan D. Kamenskikh,

Igor O. Kolbenev,

Saratov State Medical University

Alexey N. Bashkatov, Elina A. Genina, Artem A. Aratov,Anastasiya M. Parkheychuk, Valery V. Tuchin

Department of Optics and Biophotonics of Saratov State University

e-mail: [email protected]


______________________________________________


__________________________________________________


Motivation:Motivation:

It is well known that diagnostics and treatment of many diseases of human eye is connected with monitoring of glucose content. However, in spite of numerous investigations dealing with the transport of the metabolite within biological tissue, the problem of monitoring of structural changes of eye cornea under action of glucose has not been studied in detail.

Goal of the study is investigation of influence of 40% aqueous glucose solution on stuctural changes in human eye cornea at the noninflamation cornea edema after post-surgical phacoemulsification of aging cataract

It is well known that diagnostics and treatment of many diseases of human eye is connected with monitoring of glucose content. However, in spite of numerous investigations dealing with the transport of the metabolite within biological tissue, the problem of monitoring of structural changes of eye cornea under action of glucose has not been studied in detail.

Goal of the study is investigation of influence of 40% aqueous glucose solution on stuctural changes in human eye cornea at the noninflamation cornea edema after post-surgical phacoemulsification of aging cataract


______________________________________________


__________________________________________________


Materials and Methods: in vitro study

The study has been performed with ten samples of

rabbit cornea immersed in solutions with different pH in the

range from 3 to 9. The buffer solutions have been prepared

using sodium citrate (C6H5O7Na25*5H2O), citric acid

(C6H8O7*H2O), (Na2HPO4), (KH2PO4), sodium carbonate

(Na2CO3), and sodium bicarbonate (NaHCO3).

pH of these solutions (3.07, 4.04, 4.91, 5.5, 5.91, 7.01,

8.59, 8.99) have been measured by pH-meter "HANNA"

(Portugal) using standard procedure.

The study has been performed with ten samples of

rabbit cornea immersed in solutions with different pH in the

range from 3 to 9. The buffer solutions have been prepared

using sodium citrate (C6H5O7Na25*5H2O), citric acid

(C6H8O7*H2O), (Na2HPO4), (KH2PO4), sodium carbonate

(Na2CO3), and sodium bicarbonate (NaHCO3).

pH of these solutions (3.07, 4.04, 4.91, 5.5, 5.91, 7.01,

8.59, 8.99) have been measured by pH-meter "HANNA"

(Portugal) using standard procedure.


______________________________________________


__________________________________________________


Materials and Methods: in vitro study

For investigation of hydration degree, the samples have been

weighed using electronic balance (SCIENTECH, SA210) with the

precision of 1 mg. The tissue samples have been immersed into

cuvettes with the buffer solutions. Every 10 min during 2 hours, each

sample has been taken out from the solution and weighted. The

hydration degree of the cornea samples has been calculated using the

equation:

For investigation of hydration degree, the samples have been

weighed using electronic balance (SCIENTECH, SA210) with the

precision of 1 mg. The tissue samples have been immersed into

cuvettes with the buffer solutions. Every 10 min during 2 hours, each

sample has been taken out from the solution and weighted. The

hydration degree of the cornea samples has been calculated using the

equation:

where M is the mass of tissue sample and t is time. where M is the mass of tissue sample and t is time.

0,

0

M t M tH t

M t


______________________________________________


__________________________________________________


Materials and Methods: in vivo study

Cornea structure has been investigated using Heidelberg Retina Tomograph (HRT) with the Rostok Cornea Module (RCM) (Heidelberg Engineering GmbH, Germany)

Cornea structure has been investigated using Heidelberg Retina Tomograph (HRT) with the Rostok Cornea Module (RCM) (Heidelberg Engineering GmbH, Germany)


______________________________________________


__________________________________________________



Cornea thickness has been measured with Topcon 3D OCT-1000 Optical coherence Tomograph (OCT) (Japan)

Cornea thickness has been measured with Topcon 3D OCT-1000 Optical coherence Tomograph (OCT) (Japan)


______________________________________________


__________________________________________________



Sixty volunteers were enrolled for the study. The subjects were divided into two groups. The I group included thirty volunteers (21 male and 9 female at the age from 56 to 72 years) with post-surgical edema of cornea after phacoemulsification of aging cataract. The II (control) group included thirty health volunteers (16 male and 14 female at the age from 22 to 30 years).

For all persons the study of corneal structure and thickness was performed before the instillation of 40%-glucose solution (2 drops 0.05 ml = 0.1 ml) and in 20 and 40 minutes after the procedure with HRT and OCT

Sixty volunteers were enrolled for the study. The subjects were divided into two groups. The I group included thirty volunteers (21 male and 9 female at the age from 56 to 72 years) with post-surgical edema of cornea after phacoemulsification of aging cataract. The II (control) group included thirty health volunteers (16 male and 14 female at the age from 22 to 30 years).

For all persons the study of corneal structure and thickness was performed before the instillation of 40%-glucose solution (2 drops 0.05 ml = 0.1 ml) and in 20 and 40 minutes after the procedure with HRT and OCT


______________________________________________


__________________________________________________


Results: in vitro studyResults: in vitro study

3 4 5 6 7 8 9 10 11-0.2

0.0

0.2

0.4

0.6

0.8

1.0

1.2

Hyd

ratio

n de

gree

(H

)

pH

Cornea (t = 10 min) Cornea (t = 20 min) Cornea (t = 60 min) Gelatin hydration

H(t) = (M(t) - M(t = 0)) / M(t = 0)

Swelling

Shrinkage

Initial state

The comparison of hydration degree of rabbit cornea and collagen (gelatin) under action of solutions with different pH measured in different time. M is the mass of the sample; t is the time of action. The data about gelatin swelling have been obtained from Lykov A.V. Transport phenomena in capillaro-porous media / Moscow, 1954


______________________________________________


__________________________________________________


Results: in vivo studyResults: in vivo study

The typical OCT images obtained from cornea in normal state under action of 40%-glucose solution. (a) – before glucose instillation; (b) – after 20 min; (c) – after 40 min

a b

c


______________________________________________


__________________________________________________



The thickness of human cornea measured in vivo by OCT under action of 40%-glucose solution (pH = 3.5) in different time

0 20 40500

600

700

800

900

1000

1100

Thi

ckne

ss, m

Time, min

Cornea in normal state Cornea with edema

0 20 400.4

0.6

0.8

1.0

Hyd

ratio

n d

eg

ree

Time, min

The hydration degree of human cornea with edema measured in vivo under action of 40%-glucose solution (pH = 3.5) in different time


______________________________________________


__________________________________________________



The typical HRT images obtained from corneum epithelium in normal state under action of 40%-glucose solution. (a) – before glucose instillation; (b) – after 20 min; (c) – after 40 min

The typical HRT images obtained from corneum epithelium with edema under action of 40%-glucose solution. (a) – before glucose instillation; (b) – after 20 min; (c) – after 40 min

a b c


______________________________________________


__________________________________________________



The typical HRT images obtained from corneum endothelium in normal state under action of 40%-glucose solution. (a) – before glucose instillation; (b) – after 40 min

The typical HRT images obtained from corneum endothelium with edema under action of 40%-glucose solution. (a) – before glucose instillation; (b) – after 40 min

a b


______________________________________________


__________________________________________________



0 20 400

2

4

6

8

10

12

Dia

me

ter,

m

Time, min


The diameter of the corneal epithelial cells measured in vivo by HRT under action of 40%-glucose solution in different time

The diameter of the corneal endothelium cells measured in vivo by HRT under action of 40%-glucose solution in different time

0 20 40 600

4

8

12

16

20

24

Dia

met

er, m

Time, min



______________________________________________


__________________________________________________


Russian Federation governmental contacts 02.740.11.0770, and 02.740.11.0879

Russian Federation governmental contacts 02.740.11.0770, and 02.740.11.0879

Grant #224014 Network of Excellence for Biophotonics (PHOTONICS4LIFE) of the Seventh Framework Programme of Commission of the European Communities

Grant #224014 Network of Excellence for Biophotonics (PHOTONICS4LIFE) of the Seventh Framework Programme of Commission of the European Communities

Acknowledgement:

The authors thank Dr. Alexander B. Pravdin (Department of Optics and Biophotonics of Saratov State University) for the help in preparation of the buffer solutions

ocular drugs influence on the structure of ocular tissues

Documents