PVA缩甲醛吸水海绵在无创宫腔液取样和RNA检测中的安全性与可行性  

Safety and feasibility of PVA formaldehyde absorbent sponge in noninvasive uterine fluid sampling and RNA sequencing

作　　者：黄曦 李艳萍[2] 李丽娅[3] 何爱桦 HUANG Xi;LI Yanping;LI Liya;HE Aihua(Department of Reproductive Medicine,Third Xiangya Hospital,Central South University,Changsha 410013;Department of Reproductive Medicine,Xiangya Hospital,Central South University,Changsha 410008;Institute of Powder Metallurgy,Central South University,Changsha 410017,China)

机构地区：[1]中南大学湘雅三医院生殖医学中心,长沙410013 [2]中南大学湘雅医院生殖医学中心,长沙410008 [3]中南大学粉末冶金研究院,长沙410017

出　　处：《中南大学学报（医学版）》2022年第11期1504-1511,共8页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(8187061497)。

摘　　要：目的:宫腔液RNA可用作检测子宫内膜容受性,但目前仍没有无创取样方法。聚乙烯醇(polyvinyl alcohol,PVA)缩甲醛吸水海绵作为一种医用生物吸水海绵,具有良好的吸水性和亲生物性,可用于开发新型无创宫腔液取样器。本研究旨在探讨PVA缩甲醛吸水海绵对子宫内膜上皮细胞的毒性及对宫腔液RNA测序(RNA sequencing,RNA-Seq)的影响。方法:使用PVA缩甲醛吸水海绵的细胞为实验组,包括0.005%、0.01%及0.02%(w/v)的悬液组以及0.01%、0.05%及0.1%(v/v)的浸提液组,对照组仅添加完全培养基,空白组无添加。采用体外细胞毒性实验测量每组细胞的存活率。招募2019年11月至2020年1月在中南大学湘雅医院生殖医学中心进行体外受精术的8例患者,采集患者的宫腔液,实验组将宫腔液用无菌PVA缩甲醛吸水海绵吸入后再置入RNA-later保存液中,对照组直接放入等量RNA-later保存液中,随后进行RNA-Seq及数据分析。结果:体外细胞毒性实验结果显示:0.005%浓度悬液组各培养时间的细胞存活率差异无统计学意义(P=0.255);0.01%和0.02%浓度悬液组在12 h内各培养时间的细胞存活率差异无统计学意义(均P>0.05),24 h与0 h相比细胞存活率下降(分别P<0.01,P<0.05)。在同一培养时间下,3个浓度梯度的悬液组细胞存活率均比对照组低(均P<0.05),其中0.005%浓度悬液组培养至24 h时细胞存活率下降幅度小于30%,0.01%浓度悬液组在12 h时细胞存活率下降幅度大于30%,0.02%浓度悬液组在0 h时下降幅度大于30%。0.01%浓度浸提液组在6 h内各培养时间的细胞存活率差异无统计学意义(均P>0.05),12 h、24 h与0 h组相比细胞存活率下降(均P<0.01);0.05%浓度浸提液组在12 h内各培养时间的细胞存活率差异无统计学意义(均P>0.05),24 h与0 h相比存活率下降(P<0.05);0.1%浓度浸提液组中各培养时间的细胞存活率差异无统计学意义(P=0.082)。在同一培养时间下,0.01%浓度浸提液组在Objective:Uterine fluid RNA can be used as a test for endometrial receptivity,but there is still no noninvasive sampling method available.The polyvinyl alcohol(PVA)formaldehyde absorbent sponge,a medical bio-absorbent sponge with good water absorption and biophilic properties,can be used to develop a new noninvasive endometrial fluid sampler.This study aims to investigate the toxicity of PVA acetal absorbent sponges on endometrial epithelial cells and its effect on RNA sequencing(RNA-Seq).Methods:The experimental group using PVA formaldehyde absorbent sponge was prepared into 0.005%,0.01%and 0.02%(w/v)suspension,and 0.01%,0.05%and 0.1%(v/v)extract groups.The control group was only the complete culture medium.Nothing was added to the blank group.In vitro cytotoxicity assay was used to evaluate the survival rate of cells.Eight patients underwent in vitro fertilization treatment in the Reproductive Center of Xiangya Hospital,Central South University from November 2019 to January 2020.The uterine fluid of each patient was aspirated.The experimental group was inhaled with sterile PVA formaldehyde absorbent sponge and then immersed RNA-later solution.The control group was directly injected into the same amount of RNA-later solution.RNAseq and data analysis was performed later.Results:The vitro cytotoxicity assay showed that in suspension groups,there was no significance difference in cell survival between different co-culture time in 0.005%group(P=0.255).In the 0.01%and 0.02%group,there was no difference at each incubation time within 12 h(all P>0.05),but the cell survival rate was decreased at 24 h compared with 0 h(P<0.01,P<0.05).At the same co-culture time,the cell survival of the 3 concentration gradient groups were significantly lower than that of the control group(all P<0.05).The cell viability of the 0.005%concentration group was decreased less than 30%at 24 h,the 0.01%concentration group decreased more than 30%at 12 h,and the 0.02%concentration group was decreased more than 30%at 0 h.For extract groups,there wa

关 键 词：PVA缩甲醛吸水海绵 宫腔液 无创取样 RNA测序 

分 类 号：R711[医药卫生—妇产科学]