Korean J Ophthalmol.  2012 Aug;26(4):290-296. 10.3341/kjo.2012.26.4.290.

Safety of Bevacizumab on Extraocular Muscle in a Rabbit Model

• Jung JH
• Lee JH
• Lee JE
• Choi HY

• KMID: 1387397
• DOI: http://doi.org/10.3341/kjo.2012.26.4.290

Abstract

PURPOSE
The purpose of this study was to investigate the myotoxicity of bevacizumab on extraocular muscles in a rabbit model.
METHODS
Thirty New Zealand white rabbits were used for this study. The animals were evenly divided into two groups. In the first group, 15 rabbits were treated with intramuscular injections of bevacizumab (1.25 mg/0.05 mL) in the right superior rectus muscle and normal saline solution (0.05 mL) in the left superior rectus muscle. In the second group, 15 rabbits were treated with subconjunctival injections of bevacizumab (2.5 mg/0.1 mL) in the right superior subconjunctival area and normal saline solution (0.1 mL) in the left superior subconjunctival area. Five rabbits in each group were sacrificed at one day, two weeks and four weeks after the injections. Extraocular muscle samples were prepared for light microscopic (LM) and electron microscopic (EM) examination. Degrees of acute inflammation were evaluated via CD-11b immunohistochemistry, and global muscle change was investigated using hematoxylin and eosin stains. Intensity of fibrosis was evaluated using Masson trichrome stains, and ultrastructural changes were observed on EM.
RESULTS
We observed no significant inflammatory cell infiltration, muscle necrosis or fibrotic change in treated and control eyes. EM findings revealed no significant damage to muscle or vascular tissue after bevacizumab injection.
CONCLUSIONS
We found no signs of extraocular muscle toxicity after LM and EM intramuscular and subconjunctival bevacizumab injections in a rabbit model.

Keyword

Bevacizumab; Extraocular muscle; Myotoxicity

MeSH Terms

Angiogenesis Inhibitors/*administration & dosage/toxicity
Animals
Antibodies, Monoclonal, Humanized/*administration & dosage/toxicity
Conjunctiva/drug effects
Injections
Oculomotor Muscles/*drug effects
Rabbits

Figure

• 

  Fig. 1 One day following muscle injection. (A) Microscopic findings revealed no remarkable inflammatory cell infiltration in a rabbit after receiving bevacizumab injection. (B) The same degree of inflammatory cells is present in the control eye (H&E, ×200). (C) CD-11b antibody immunostaining of the extraocular muscle. Note the lack of remarkable staining of CD-11b-positive inflammatory cells (which stain a brown or black color, and indicates the presence of monocytes, macrophages and neutophils) infiltration in the muscle layer of a bevacizumab injection in the rabbit. (D) Control group showing the no significant degree of CD-11b-positive cellular response (×200).

• 

  Fig. 2 Four weeks after muscle injection of bevacizumab (A) and normal saline (B). The muscle shows a uniform diameter, even staining and unremarkable fibrosis compared to the control eye (Masson's trichrome, ×100).

• 

  Fig. 3 Subconjunctival injection group. (A) One day following bevacizumab injection in the subconjunctival space. Microscopic findings reveal regularly arranged individual muscle fibers surrounded by thin fibrous connective tissues and blood vessels (H&E, ×200). (B) Normal appearance in a normal saline injection eye (H&E, ×200) one day after injection. (C,D) Four weeks after bevacizumab and normal saline injections, a small amount of collagen fibers and minimal fibrosis are present in surrounding tissue (Masson's trichrome, ×100).

• 

  Fig. 4 Electron micrograph of extraocular muscle after bevacizumab muscle injection. (A) Longitudinal section of myofiber one day following bevacizumab muscle injection. Normal, ultra structural appearance of muscle fiber with subsarcolemmal nuclei is observed (black arrow). The distinct and intact banding patterns of sarcomeres and the Z-band are visible (×3,000). (B) Transverse section of the muscle and capillaries one day after bevacizumab muscle injection. Endothelial cells of the capillaries exhibit normal cytology, and the capillaries show intact structural integrity (black arrow head) (×3,000). (C) Longitudinal section of a myofiber four weeks after bevacizumab muscle injection. Muscle fibers and mitochondrial membranes (white arrow) are seen (×3,000). (D) Four weeks after bevacizumab injection, capillary endothelium and connective tissue of the muscle layer are intact. Dense and regular, uniform arrayed muscle fibers are seen (white arrow head) (×3,000).

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