N V Kosheleva, I V Ilina, K V Kozhina, I M Zurina, A E Roskova, A A Gorkun, A V Ovchinnikov, M B Agranat, S G Morozov, I N Saburina
{"title":"[Cellular Model Based on Laser Microsurgery of Cell Spheroids to Study the Repair Process].","authors":"N V Kosheleva, I V Ilina, K V Kozhina, I M Zurina, A E Roskova, A A Gorkun, A V Ovchinnikov, M B Agranat, S G Morozov, I N Saburina","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>In this study, modern techniques of laser microsurgery of cell spheroids have been used to develop\na new simple, reproducible model for studying the mechanisms of repair and regeneration in vitro. Nanosecond\nlaser pulses were applied to perform a microdissection of the outer and the inner zones of the spheroids\nfrom dermal fibroblasts. To achieve effective dissection and preservation of spheroid viability, the optimal\nparameters were chosen: 355 nm wavelength, 100 Hz frequency, 2 ns pulse duration, laser pulses in the range\nof 7–9 μ J. After microdissection, we observed injury of the spheroids : the edges of the wound surface opened\nand the angular opening reached a value of more than 180°. As early as during the first hour after spheroid\nmicrodissection with laser radiation, the surviving cells changed their shape: cells on the spheroid surface and\ndirectly in the damaged area became rounded. One day after microdissection, the structure of the spheroids\nbegan to partially recover, the cells in the surface layers began to take the original flattened shape; debris of\ndead damaged cells and their fragments was gradually cleared from the spheroid composition. In the proposed\nmodel, the first data on stimulation of structure recovery of injured spheroids from dermal fibroblasts with a\nP199 synthetic polypeptide, which is used in cosmetology for the initiation of antiaging and regenerative\neffects in the skin, were received. After microdissection, recovery of the spheroids structure with a few surface\nlayers of flattened imbricated arranged cells and polygonal cells of the inner zone in the presence of P199 peptide\nwas faster than in the control group, and was completed within 7 days, presumably due to the remodeling\nof the survived cells.</p>","PeriodicalId":19673,"journal":{"name":"Ontogenez","volume":"48 1","pages":"63-72"},"PeriodicalIF":0.0000,"publicationDate":"2017-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Ontogenez","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
In this study, modern techniques of laser microsurgery of cell spheroids have been used to develop
a new simple, reproducible model for studying the mechanisms of repair and regeneration in vitro. Nanosecond
laser pulses were applied to perform a microdissection of the outer and the inner zones of the spheroids
from dermal fibroblasts. To achieve effective dissection and preservation of spheroid viability, the optimal
parameters were chosen: 355 nm wavelength, 100 Hz frequency, 2 ns pulse duration, laser pulses in the range
of 7–9 μ J. After microdissection, we observed injury of the spheroids : the edges of the wound surface opened
and the angular opening reached a value of more than 180°. As early as during the first hour after spheroid
microdissection with laser radiation, the surviving cells changed their shape: cells on the spheroid surface and
directly in the damaged area became rounded. One day after microdissection, the structure of the spheroids
began to partially recover, the cells in the surface layers began to take the original flattened shape; debris of
dead damaged cells and their fragments was gradually cleared from the spheroid composition. In the proposed
model, the first data on stimulation of structure recovery of injured spheroids from dermal fibroblasts with a
P199 synthetic polypeptide, which is used in cosmetology for the initiation of antiaging and regenerative
effects in the skin, were received. After microdissection, recovery of the spheroids structure with a few surface
layers of flattened imbricated arranged cells and polygonal cells of the inner zone in the presence of P199 peptide
was faster than in the control group, and was completed within 7 days, presumably due to the remodeling
of the survived cells.