CAJ | 학술논문

目的：观察bevacizumab对人Tenon囊成纤维细胞迁移能力的影响,探讨青光眼术后滤过泡瘢痕化的对策。方法：采用从陕西省人民医院中心实验室细胞库中取出冻存的人Tenon囊成纤维细胞,采用细胞复苏法,遵循无菌原则,进行常规培养。创伤划痕试验：待细胞融合度达到80%时在单层细胞表面划出一条无细胞的刮除带,空白对照组加入不含血清的DMEM培养液,bevacizumab处理组加入 bevacizumab 浓度为1 mg/mL 不含血清的DMEM培养液,分时段(0,24,48,72h)观察并测量划痕宽度。结果：人Tenon囊成纤维传代细胞显微镜下观察呈长梭形,细胞核位于细胞的中部,核较大,胞浆丰富,生长时呈漩涡状排列走形,增殖能力强,符合成纤维细胞的一般形态。冻存和复苏后细胞的形态结构和生物学特点维持不变。细胞划痕试验显示：0h时,两组细胞划痕初始宽度相等；24 h 时两组细胞迁移距离基本一致；48 h 时bevacizumab处理组细胞迁移距离明显小于空白对照组；72 h时空白对照组划痕基本愈合, bevacizumab处理组细胞迁移距离较48 h无明显变化,且细胞死亡数目较多。结论：利用细胞复苏法可成功培养形态结构和生物学特性稳定的传代人Tenon囊成纤维细胞,为实验研究奠定细胞学基础。成纤维细胞本身具有较强的迁移能力,外源性bevacizumab作用可以明显抑制细胞的迁移,作用时间过长时,会造成细胞的过多死亡。抗新生血管药物对成纤维细胞的迁徙具有一定的抑制作用,未来很有可能成为眼科临床抗击青光眼术后滤过泡瘢痕化的重要手段。
목적：관찰bevacizumab대인Tenon낭성섬유세포천이능력적영향,탐토청광안술후려과포반흔화적대책。방법：채용종합서성인민의원중심실험실세포고중취출동존적인Tenon낭성섬유세포,채용세포복소법,준순무균원칙,진행상규배양。창상화흔시험：대세포융합도체도80%시재단층세포표면화출일조무세포적괄제대,공백대조조가입불함혈청적DMEM배양액,bevacizumab처리조가입 bevacizumab 농도위1 mg/mL 불함혈청적DMEM배양액,분시단(0,24,48,72h)관찰병측량화흔관도。결과：인Tenon낭성섬유전대세포현미경하관찰정장사형,세포핵위우세포적중부,핵교대,포장봉부,생장시정선와상배렬주형,증식능력강,부합성섬유세포적일반형태。동존화복소후세포적형태결구화생물학특점유지불변。세포화흔시험현시：0h시,량조세포화흔초시관도상등；24 h 시량조세포천이거리기본일치；48 h 시bevacizumab처리조세포천이거리명현소우공백대조조；72 h시공백대조조화흔기본유합, bevacizumab처리조세포천이거리교48 h무명현변화,차세포사망수목교다。결론：이용세포복소법가성공배양형태결구화생물학특성은정적전대인Tenon낭성섬유세포,위실험연구전정세포학기출。성섬유세포본신구유교강적천이능력,외원성bevacizumab작용가이명현억제세포적천이,작용시간과장시,회조성세포적과다사망。항신생혈관약물대성섬유세포적천사구유일정적억제작용,미래흔유가능성위안과림상항격청광안술후려과포반흔화적중요수단。
Abstract? AlM: To investigate the inhibition effect of bevacizumab on human Tenon capsule fibroblasts ( HTFs ) and discuss the countermeasures of bleb scarringscarring in glaucoma surgery countermeasures.? METHODS: Adopted cell recovery method and followed the aseptic principles, we performed the culture of HTFs which came from the Central Laboratory of Shaanxi People's Hospital cell library. Wound Healing assay:We scraped a cell-free zone on the cell surface when the cells reached confluence at 80%. The control group was added to serum-free DMEM medium. The HTFs of bevacizumab group were stimulated with 1mg/mL concentrations without DEME for 0, 24, 48, and 72h. The scratch width was observed and measured.? RESULTS: HTFs were long fusiform shape under microscope, the nucleus is in the center of the cell with larger nucleus, abundant cytoplasm, were arranged in a whorled growth out of shape, strong ability to proliferate, conform to general forms of fibroblast. The morphological and biological characteristics of cells after cryopreservation and resuscitation remain unchanged. Wound healing assay: 0h, equal to the initial width of the two groups, 24h when the migration distance of the two groups of cells are basically the same, 48h when the control group cell migration distance is greater than that in the bevacizumab processing group, 72h when the control group scratches basichealing, bevacizumab treated cells migrate closer than 48h no significant change, and a lot of cells died.?CONCLUSlON:Cell recovery method can successfully cultured HTFs, which was stability on morphology and biological properties, laying the cellular basis for experimental research. Fibroblast itself has a strong ability to migrate, outside - derived bevacizumab can inhibit HTFs migration evidently and it will cause excessive cell death when. Bevacizumab has certain extent inhibitory effect on HTFs migration, and it is likely to become one of the important drugs for creating bleb scarring after glaucoma surgery in the future.