Intracellular calcium response and adhesiveness of chick embryonic retinal neurons to photoelectric dye-coupled polyethylene films as prototypes of retinal prostheses.
Photoelectric dyes, which absorb light and convert photon energy to electric potentials, were shown to stimulate retinal neurons in culture. One of these dyes coupled with polyethylene film surface, as a prototype of retinal prostheses, could induce intracellular calcium elevation in chick embryonic retinal tissues. In this study, we used retinal cells from chick embryos in which no photoreceptor outer segments yet developed, and assessed cell adhesiveness and response to the original and modified types of our retinal prostheses. Extruded-blown film of high density polyethylene was used as the original polyethylene film. Polyethylene film surface was modified either by coupling with type 1 collagen or by crystallization from the melt of the original polyethylene film. A photoelectric dye, 2-[2-[4-(dibutylamino)phenyl]ethenyl]-3-carboxymethylbenzothiazolium bromide, was coupled to these different types of polyethylene films through amide linkage. Retinal cells from chick embryos at the 12-day embryonic stage were cultured on the surface of dye-coupled films. The response of retinal cells to the dye-coupled films was observed by calcium imaging. Cell types, either neurons or glial cells, were detected by immunocytochemical staining and also observed by scanning electron microscopy (SEM). Intracellular calcium elevation was observed in cells, including both neurons and glial cells, on the surface of the original dye-coupled film, collagen-coated dye-coupled film, and dye-coupled recrystallized film, in contrast with no response of cells cultured on the dye-uncoupled plain film. Retinal neurons grew upon colonies of glial cells settling on the surface of all different types of the films. The number of retinal neurons on the collagen-coated dye-coupled film and dye-coupled recrystallized film was significantly larger than the number of neurons on the dye-uncoupled plain film (P = 0.0004, analysis of variance; P < 0.05, Tukey-Kramer test). With SEM, Muller cell-like large cells covered the original dye-coupled film and collagen-coated dye-coupled film, while neuron-like small cells settled directly on the dye-coupled recrystallized film and also formed large colonies on Muller-like cells. Retinal neurons and glial cells were cultured directly on the different types of dye-coupled polyethylene films. Photoelectric dye-coupled polyethylene films could be used as prototypes of retinal prostheses.
Uji
Akihito
A
Department of Ophthalmology, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University, Okayama City, Japan.
Matsuo
Toshihiko
T
Uchida
Tetsuya
T
Shimamura
Kaoru
K
Ohtsuki
Hiroshi
H
D000814:Aniline Compounds; D000818:Animals; D001705:Bioprosthesis; D002118:Calcium / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques; D002642:Chick Embryo; D020099:Coated Materials, Biocompatible; D003094:Collagen; D003460:Crystallization; D042541:Intracellular Space / Q000378:metabolism*; D009474:Neurons / Q000502:physiology*; D020959:Polyethylene; D012160:Retina / Q000166:cytology* / Q000196:embryology; D013844:Thiazoles
C501867:2-(2-(4-(dibutylamino)phenyl)ethenyl)-3-carboxymethylbenzothiazolium; D000814:Aniline Compounds; D020099:Coated Materials, Biocompatible; D013844:Thiazoles; D020959:Polyethylene; D003094:Collagen; D002118:Calcium