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

• Published in: PloS one Vol. 13; no. 12; p. e0209996
• Main Authors: Takahashi, Kosuke, Morizane, Yuki, Hisatomi, Toshio, Tachibana, Takashi, Kimura, Shuhei, Hosokawa, Mio Morizane, Shiode, Yusuke, Hirano, Masayuki, Doi, Shinichiro, Toshima, Shinji, Araki, Ryoichi, Matsumae, Hiroshi, Kanzaki, Yuki, Hosogi, Mika, Yoshida, Atsushi, Sonoda, Koh-Hei, Shiraga, Fumio
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 31.12.2018; Public Library of Science (PLoS)
• Subjects: Animals; Apoptosis; Biology and Life Sciences; Dentistry; Disruption; DNA nucleotidylexotransferase; Electron microscopy; Epithelium; Eye; Eye (anatomy); Gene therapy; High pressure; Injection; Injections, Intraocular; Laboratory animals; Light microscopy; Macaca fascicularis; Macular degeneration; Male; Medicine; Medicine and Health Sciences; Microscopy, Electron, Transmission; Microstructure; Monkeys; Monkeys & apes; Optical Coherence Tomography; Pharmaceutical sciences; Photoreceptors; Pressure; R&D; Research & development; Research and Analysis Methods; Retina; Retinal pigment epithelium; Retinal Pigment Epithelium - ultrastructure; Saline solutions; Social Sciences; Stem cells; Surgery; Tomography; Tomography, Optical Coherence; Transmission electron microscopy; University graduates; Vitrectomy; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0209996

To investigate the influence of subretinal injection pressure on the microstructure of the retina in a monkey model. 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. 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. 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.