Comparison of Vitreolytic Effect in Rabbit Eyes: Plasmin, Hyaluronidase, and Their Mixtures

Moo Sang Kim; Sang Woong Moon; Eung Suk Kim; Seung Young Yu; Hyung Woo Kwak

doi:10.3341/jkos.2009.50.6.911

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Kim, Moon, Kim, Yu, and Kwak: Comparison of Vitreolytic Effect in Rabbit Eyes: Plasmin, Hyaluronidase, and Their Mixtures

Original Article

J Korean Ophthalmol Soc 2009;50(6):911-918.

Published online: 7 September 2009

DOI: https://doi.org/10.3341/jkos.2009.50.6.911

Comparison of Vitreolytic Effect in Rabbit Eyes: Plasmin, Hyaluronidase, and Their Mixtures

Moo Sang Kim, MD¹, Sang Woong Moon, MD², Eung Suk Kim, MD³, Seung Young Yu, MD¹, Hyung Woo Kwak, MD¹

¹Department of Ophthalmology, KyungHee University Hospital, Seoul, Korea

²Department of Ophthalmology, Inje University College of Medicine, Seoul Paik Hospital, Seoul, Korea

³Department of Ophthalmology, Chung-Ang University College of Medicine, Seoul, Korea

Address reprint requests to Hyung Woo Kwak, MD, PhD Department of Ophthalmology, KyungHee University Hospital #1 Hoegi-dong, Dongdaemun-gu, Seoul 130-702, Korea Tel: 82-2-958-8455, Fax: 82-2-966-7340, E-mail: hwkwak@khmc.or.kr

Received 25 August 2008 Accepted 2 March 2009

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

The aim of the present study was to quantify and compare the vitreolytic effect of plasmin, hyaluronidase, and a combination of the two.

Methods

Thirty-six rabbits were randomized into 3 groups: (A) twelve rabbits had an intravitreal injection of plasmin 1 U with hyaluronidase 10 U/0.1 mL into the right eye, (B) twelve rabbits had an injection of plasmin alone (1 U/0.1 mL), and (C) twelve rabbits had an injection of hyaluronidase alone (10 U/0.1 mL). The left eye of each rabbit was used as control, which was injected with 0.1 mL phosphate buffered saline (PBS). The eyes were enucleated 1 hour and 24 hours after injection. The volume of fluid-type vitreous and gel-type vitreous was measured with a micropipette using the melting point as the difference. Statistical analysis was performed and light microscopy was used to assess potential damage to the retinal tissue.

Results

The volume of remaining gel-type vitreous was measured as 52.5%, 60.3%, 59.2%, and 76.5% after 1 hour enucleation and as 44.6%, 56.7%, 56.1%, and 74.7%, after 24 hours enucleation in group A, B, C, and control group, respectively. Group A, B, and C showed statistically significant differences against the control group. Group A (plasmin with hyaluronidase) showed less remaining gel-type vitreous volume than a single injection of plasmin or hyaluronidase alone.

Conclusions

Intravitreal injection of plasmin with hyaluronidase showed more vitreolytic effect than a single injection of plasmin or hyaluronidase alone. The enzyme may be useful in liquefying the vitreous, and may be a useful biochemical adjunct to vitrectomy.

Keywords: Hyaluronidase, Intravitreal injection, Pharmacologic vitreolysis, Tissue plasminogen activator

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Figure 1.

Photographs of the experimental process. (A, B and C) Peeling the sclera, choroid, retina tissue of frozen rabbit eyeball. (D, E) Removal of the cornea, aqueous humor, iris and lens of frozen eyeball. (F) Separated frozen vitreous, lens and iris respectively (Blue asterisk: vitreous green asterisk: lens white asterisk: iris). (G, H and I) Melting the vitreous at room temperature (red arrow: gel type vitreous yellow arrow: fluid type vitreous).

Figure 2.

Comparison of the plasmin and hyaluronidase injection group with the control group. At 1 hour after enucleation, volume of gel type vitreous per whole vitreous was 52.5% and 76.5% in group A and the control group, respectively. At 24 hours after enucleation, volume of gel type vitreous per whole vitreous was 44.6% and 74.7%, respectively. Volume of gel type vitreous per whole vitreous showed statistically significant difference between group A and the control group (1 hour: p=0.000; 24 hours: p=0.000).

Figure 3.

Comparison of the plasmin injection group with the control group. At 1 hour after enucleation, volume of gel type vitreous per whole vitreous was 60.3% and 76.5% in group B and the control group, respectively. At 24 hours after enucleation, volume of gel type vitreous per whole vitreous was 56.7% and 74.7%, respectively. volume of gel type vitreous per hole vitreous showed statistically significant difference between group A and the control group (1 hour: p=0.000; 24 hours: p=0.000).

Figure 4.

Comparison of the hyaluronidase injection group with the control group. At 1 hour after enucleation, volume of gel type vitreous per whole vitreous was 69.2% and 76.5% in group C and the control group, respectively. At 24 hours after enucleation, volume of gel type vitreous per whole vitreous was 56.1% and 74.7%, respectively. volume of gel type vitreous per whole vitreous showed statistically significant difference between group A and the control group (1 hour: p=0.000; 24 hours: p=0.000).

Table 1.

The mean volume of remained gel type vitreous

	Mean volume of gel type vitreous
	I hour after enucleation	24 hours after enucleation	p-value^П
Group A^*	52.5±2.3%	44.6±2.1%	p=0.018
Group B^†	60.3±2.1%	56.7±2.4%	p=0.000
Group C^‡	59.2±3.6%	56.1±2.0%	p=0.179
Control^§	76.5±2.5%	74.7±2.4%	

^* Group A=plasmin 1 U+hyaluronidase 10 U

^† Group B=plasmin 1 U

^‡ Group C=hyaluronidase 10 U

^§ Control=phosphate buffered saline (PBS) 0.1 mL

^П Comparison between 1 hour after injection and 24 hours after injection.

Table 2.

Comparison of the mean volume of remained gel type vitreous between groups (at 24 hours after injection)

		Mean volume of gel type vitreous (%)		p value
Group A^*	Group B	44.6±2.1	56.7±2.4	0.000
	Group C		56.1±2.0	0.000
	Control^§		74.7±2.4	0.000
Group B^†	Group C	56.7±2.4	56.1±2.0	0.647
Group B^†	Control	56.7±2.4	74.7±2.4	0.000
Group C^‡	Control	56.1±2.0	74.7±2.4	0.000

^* Group A=plasmin 1 U+hyaluronidase 10 U

^† Group B= plasmin 1 U

^‡ Group C=hyaluronidase 10 U

^§ Control= phosphate buffered saline (PBS) 0.1 mL.

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