The influence of subretinal injection pressure on the microstructure of the monkey retina

Kosuke Takahashi, Yuki Morizane, Toshio Hisatomi, Takashi Tachibana, Shuhei Kimura, Mio Morizane Hosokawa, Yusuke Shiode, Masayuki Hirano, Shinichiro Doi, Shinji Toshima, Ryoichi Araki, Hiroshi Matsumae, Yuki Kanzaki, Mika Hosogi, Atsushi Yoshida, Koh Hei Sonoda, Fumio Shiraga

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Abstract

Purpose To investigate the influence of subretinal injection pressure on the microstructure of the retina in a monkey model. Methods After vitrectomy, balanced salt solution was injected subretinally into one eye each of four cynomolgus monkeys while controlling the injection pressure. Initially, a pressure of 2 psi was used, and this was gradually increased to determine the minimum required pressure. Subsequent injections were performed at two pressures: minimum (n = 13) and high (n = 6). To compare the influence of these injection pressures on retinal structure, optical coherence tomography (OCT) was performed before surgery and every week afterwards. The monkeys were euthanized and their eyes were enucleated at 1 or 6 weeks after the injections. The eyes were processed for light microscopy and transmission electron microscopy (TEM) as well as for TdT-mediated dUTP nick end labeling. Results The minimum pressure required to perform subretinal injection was 6 psi. After injection at this pressure, both OCT and microscopy showed that the retinal structure was well-preserved throughout the experimental period at all injection sites. Conversely, after injection at high pressure (20 psi) OCT images at all injection sites showed disruption of the ellipsoid zone (EZ) after 1 week. Microscopy indicated damage to the photoreceptor outer segment (OS) and stratification of the retinal pigment epithelium (RPE). After 6 weeks, OCT demonstrated that the EZ had become continuous and TEM confirmed that the OS and RPE had recovered. Photoreceptor apoptosis was absent after subretinal injection at both pressures. Conclusions The retinal damage caused by subretinal injection increases depending on pressure, indicating that clinicians should perform subretinal injection at pressures as low as possible to ensure safety.

Original languageEnglish
Article numbere0209996
JournalPLoS One
Volume13
Issue number12
DOIs
Publication statusPublished - Dec 1 2018

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

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    Takahashi, K., Morizane, Y., Hisatomi, T., Tachibana, T., Kimura, S., Hosokawa, M. M., Shiode, Y., Hirano, M., Doi, S., Toshima, S., Araki, R., Matsumae, H., Kanzaki, Y., Hosogi, M., Yoshida, A., Sonoda, K. H., & Shiraga, F. (2018). The influence of subretinal injection pressure on the microstructure of the monkey retina. PLoS One, 13(12), [e0209996]. https://doi.org/10.1371/journal.pone.0209996