P-选择素对角膜上皮创伤修复及其中性粒细胞迁移的影响

目的观察P-选择素对角膜上皮创伤修复及其中性粒细胞迁移的影响。方法选用P-选择素基因敲除小鼠与野生型小鼠进行对比,观察角膜上皮创伤修复过程,同时用免疫组织化学方法观察中性粒细胞在角膜的迁移,并用酶联免疫吸附测定方法检测角膜中细胞因子和趋化因子的变化。结果与野生型小鼠相比,P-选择素基因敲除小鼠的角膜创伤修复延迟,向角膜迁移的中性粒细胞数量减少;中性粒细胞的迁移与趋化因子和细胞因子有密切的关系。结论P-选择素表达缺陷可导致角膜上皮创伤修复的延迟。这种延迟可能与创伤后中性粒细胞向创伤区迁移数量减少有关。     

左旋肉碱促进碱烧伤后角膜上皮修复的作用及机制

目的：探讨在角膜碱烧伤所致高渗和炎症环境中,左旋肉碱(LC)对角膜上皮修复的作用及其调控的分子机制。

方法：将96只健康C57/6J小鼠随机分为空白对照组、磷酸缓冲盐溶液(PBS)组和LC组,空白对照组不做任何处理、LC组及PBS组制备急性碱烧伤模型。自模型制作前1d,LC组给予60mmol/L LC滴眼液,PBS组给予磷酸缓冲盐溶液连续每天点眼,每天6次。建模后0h,3、7d裂隙灯显微镜下荧光素钠染色观察角膜上皮修复情况。在第3d,取眼球进行冰冻切片免疫荧光检测Ki-67、IL-1β蛋白表达情况; 提取角膜上皮全蛋白进行Western blot检测P63、NLRP3、Caspase-1的表达及STAT3的活化水平。

结果：角膜荧光素钠染色结果显示在碱烧伤后3、7d时PBS组与LC组比,残留角膜上皮缺损面积占原始缺损面积的百分比分别为(29.38±6.83)% vs(17.78±4.11)%、(14.23±4.51)% vs(4.10±2.10)%(P<0.01)。LC组比PBS组角膜上皮修复快。造模后3d,与空白对照组比,碱烧伤处理的各组小鼠角膜上皮中焦亡相关蛋白NLRP3、Caspase-1表达上调,P63表达降低,p-STAT3/STAT3水平升高,除cleaved Caspase-1空白对照组vs LC组,差异均有意义。与PBS组比,LC组的NLRP3、pro Caspase-1、cleaved Caspase-1蛋白表达降低,P63表达量上调,p-STAT3/STAT3水平升高,差异均有意义。免疫荧光显示,碱烧伤后3d,PBS组和LC组与空白对照组比IL-1β、Ki-67表达上调。与PBS组比,LC组Ki-67蛋白表达上调,IL-1β表达降低。

结论：LC可通过抑制细胞焦亡信号通路,促进STAT3信号通路的活化,促使小鼠角膜上皮干/祖细胞增生,进而促进碱烧伤后角膜上皮的修复。     

OCTA分析埃罗替尼对小鼠角膜厚度的影响

目的：通过光学相干断层扫描血管造影,对小鼠角膜上皮和角膜全层厚度进行检测并分析埃罗替尼对其的影响。

方法：随机均分20只小鼠为埃罗替尼组和PBS组,制备埃罗替尼滴眼液,分别应用埃罗替尼滴眼液和PBS,于每日8：00、12：00、16：00和20：00对2组小鼠点眼。在点眼前、点眼后1、2、3wk应用OCTA测量角膜17个区域的上皮和角膜全层厚度。

结果：点眼前埃罗替尼组与PBS组各区域角膜上皮和角膜全层厚度比较无差异(均P>0.05); 点眼后第2wk,埃罗替尼组小鼠角膜上皮和角膜全层的17个区域中M、T5、IT5、I5、IN5、N5、T6、IT6、I6、IN6、N6区域与PBS组相比均明显增厚(均P<0.05)。第3wk,埃罗替尼组小鼠角膜上皮和角膜全层厚度的17个区域与PBS组相比均明显增厚(均P<0.05); 埃罗替尼组和PBS组经处理2、3wk后,各组间角膜上皮厚度和角膜全层厚度平均值相比有差异(均P<0.05); 通过角膜上皮和角膜全层厚度平均值变化趋势分析,埃罗替尼组与PBS组均有差异(均P<0.05)。

结论：利用OCTA可以发现埃罗替尼具有使角膜上皮和角膜全层增厚的作用,且随着应用次数的增多,其效果越明显。     

地塞米松对大鼠角膜酸烧伤疗效初探

目的：探讨不同时期使用地塞米松对大鼠角膜酸烧伤的疗效。

方法：40只SD大鼠的40只角膜酸烧伤眼按随机数字法分成4组,每组10只,其中Ⅰ,Ⅱ,Ⅲ组为治疗组,分别于角膜烧伤后0～3,3～7,7～21d使用地塞米松滴眼; Ⅳ组为空白对照组。于烧伤后使用裂隙灯显微镜动态观察角膜上皮缺损直径、角膜新生血管及角膜基质混浊程度并照相,并取角膜行HE染色观察角膜各层组织的形态学变化。

结果：最终角膜修复优良情况依次为Ⅲ组,Ⅰ组,Ⅳ组,Ⅱ组; 酸烧伤后各组角膜上皮愈合速度和新生血管面积不同,不同时期的实验组与对照组间的比较有统计学意义(P<0.01)。

结论：在酸烧伤后不同时期使用地塞米松,疗效存在差异。在急性期和修复晚期使用地塞米松能够促进角膜的修复,在修复早期使用地塞米松则会阻碍角膜的修复甚至可促使角膜溶解。     

角膜前基质针刺术治疗角膜上皮细胞功能障碍的临床疗效

目的：探讨角膜前基质针刺术(ASP)治疗角膜上皮细胞功能障碍(CED)的临床疗效。

方法：选取2015-09/12于华中科技大学同济医学院附属协和医院眼科行ASP治疗的CED患者16例16眼,分别于术前、术后1、3mo观察裸眼视力、眼表疾病指数评分(OSDI)、角膜荧光染色、角膜上皮厚度、全角膜厚度、角膜上皮下树突状细胞密度、角膜内皮细胞密度、角膜上皮下神经丛密度。

结果：术后1mo,本组患者裸眼视力、上皮下神经丛密度均较术前明显改善,OSDI评分、角膜上皮厚度、全角膜厚度、角膜上皮下树突状细胞密度均较术前明显降低(P<0.05),而角膜内皮细胞密度较术前无明显变化(P>0.05)。术后3mo,裸眼视力、OSDI评分、角膜上皮厚度、全角膜厚度、角膜内皮细胞密度与术后1mo无明显差异(P>0.05),但角膜上皮下树突状细胞密度明显下降,上皮下神经丛密度明显增加(P<0.05)。

结论：ASP可以有效治疗CED及其诱发的炎症,并可修复其导致的角膜上皮下神经丛缺损。     

芥子气兔角膜损伤和角膜中央厚度变化

目的：观察芥子气兔角膜损伤病变发生发展过程中角膜上皮的愈合情况和角膜中央厚度的变化。

方法：设实验组10眼,对照组10眼,正常组20眼。实验组和对照组分别用浓度400μL/L(0.2mL)芥子气液和浓度200μL/L(0.2mL)芥子气液接触角膜4min,于1,2,3,4,5,6d; 1,2,3wk; 1,3mo行角膜白光及荧光照相,并测量角膜中央厚度。

结果：实验组的角膜上皮损伤面积1wk内由Ⅳ级减小为0级,第2,3wk; 1mo角膜上皮反复发生点片状剥脱。对照组的角膜上皮损伤面积3d内由Ⅲ级减小为0级,第2,3wk; 1mo角膜上皮仅发生针尖样的缺损。第3mo实验组和对照组角膜上皮均恢复光滑平面。角膜中央厚度第1～6d实验组明显高于对照组和正常组,差异有统计学意义(P<0.05)。第1wk～1mo实验组、对照组和正常组相比,差异无统计学意义。第3mo实验组、对照组高于正常组,但差异无统计学意义。

结论：不同浓度的芥子气液可致兔角膜上皮不同程度的损伤和角膜厚度变化,1wk内出现一过性角膜上皮层的重塑改变,1mo内完全恢复。3mo内角膜厚度基本恢复正常。     

角巩膜缘切口与角膜切口超声乳化术治疗年龄相关性白内障的疗效比较

目的：分析角巩膜缘切口与角膜切口超声乳化术对年龄相关性白内障患者术后康复的影响。

方法：选择2015-01/2017-01在我院行超声乳化术的年龄相关性白内障患者98例132眼，按手术切口位置分为角巩膜缘切口组(A组，52例74眼)与角膜切口组(B组，46例58眼)，手术前后不同时间测定两组裸眼视力、BCVA、角膜散光度的变化，统计两组上皮修复时间、切口修复时间，记录治疗前后不同时间患者BUT、泪液分泌量及角膜染色评分的变化，统计两组术后并发症发生率。

结果：两组组内、组间不同时间裸眼视力、BCVA、角膜散光度、手术源性角膜散光度、BUT、泪液分泌量、角膜染色评分均有差异(P<0.05)，A组术后不同时间裸眼视力、BCVA优于B组，角膜散光度与手术源性角膜散光度低于B组，BUT长于B组，泪液分泌量高于B组，角膜染色评分低于B组(均P<0.05)； A组术后上皮修复时间、切口修复时间均短于B组(P<0.05)； 两组术后并发症发生率无差异(P>0.05)。

结论：角巩膜缘切口下超声乳化手术可改善年龄相关性白内障患者术后视力，降低角膜散光度，减少对泪膜功能的影响，促进患者术后康复。     

自体血清联合重组人表皮生长因子对角膜创面的修复作用

目的：探讨自体血清联合重组人表皮生长因子对角膜异物剔除术后角膜创面的修复效果。

方法：选取2015-10/2016-03于我院行角膜异物剔除手术的患者92例110眼,随机分为治疗组和对照组,术后对照组患者使用重组人表皮生长因子滴眼液点眼,治疗组患者在对照组的基础上联合使用自体血清滴眼。手术前后,采用AS-OCT观察角膜创面情况。术后评估两组患者的临床疗效及泪液中TNF-α和IL-6的浓度。

结果：术后1wk治疗组患者治愈率(96.4% vs 78.2%)和术后2d角膜荧光素染色阴性率(98.2% vs 83.6%)均明显高于对照组,泪液中TNF-α和IL-6(51.47±7.83、5.29±1.67ng/L)浓度均明显低于对照组(143.62±18.12、13.95±3.44ng/L)。术后治疗组患者角膜上皮愈合时间(1.4±0.6d)较对照组(2.2±1.3d)明显缩短。术后1wk,AS-OCT图像显示治疗组修复后的角膜基质形态更加接近术前正常水平。

结论：自体血清联合重组人表皮生长因子对角膜异物剔除术后角膜创面的修复效果显著,可以明显缩短角膜创面愈合时间,减轻炎症反应,对角膜基质具有良好的修复功能。     

羊膜泪小管支架治疗去势雄兔干眼症的实验研究

目的：探究羊膜泪小管支架对手术阉割雄兔干眼症的影响。

方法：将36只成年雄性兔分组,其中A组行假阉割手术,B组假植入泪小管支架,C组植入泪小管支架。分别于植入前和植入后2、4、6wk行角膜荧光素染色(FL)和泪液分泌试验(SⅠt)检查,共聚焦显微镜观察角膜神经变化。

结果：A组角膜光滑、清晰,FL染色不明显; B组角膜粗糙,角膜上皮见点染色斑点; C组角膜较假手术组更为光滑、清晰,无明显点状染色。各组植入后2、4、6wk时FL染色评分和SⅠt比较,差异有统计学意义(均P<0.05)。共聚焦显微镜下可见A组为相对直的神经丛,量多,清晰可见; B组角膜上皮细胞下神经可见弯曲,数量有所减少; C组角膜上皮下神经相对直,与正常雄兔角膜上皮下神经相比,数量略有减少。

结论：羊膜泪小管支架可显著改善阉割去势雄兔干眼症的症状和体征,具有治疗作用。     

甘草甜素对小鼠角膜急性碱烧伤后新生血管的抑制作用

目的：研究甘草甜素(Gly)对小鼠角膜急性碱烧伤后角膜新生血管(CNV)的抑制作用。

方法：采用碱烧伤法制备小鼠碱烧伤模型60只,随机平均分为Gly组和PBS组,分别隔天结膜下注射2g/L Gly溶液或PBS溶液,共14d。裂隙灯显微镜下观察角膜炎症反应和CNV。实验结束后取角膜行HE染色法和免疫组织化学法CD34及髓过氧化物酶(MPO)染色并计算角膜微血管密度(MVD)及中性粒细胞数。

结果：造模后第7、14d Gly组CNV面积分别是4.16±0.00、7.33±0.13mm²,显著低于PBS组(7.58±0.20、9.24±0.08mm²,均P<0.05)。HE病理染色显示正常小鼠角膜结构完整,未见新生血管形成及炎症细胞浸润; Gly组角膜新生血管数量及炎症细胞浸润较少,胶原排列较规则,而PBS组角膜基质中可见大量炎症细胞浸润及新生血管。免疫组织化学CD34染色结果显示Gly组MVD为11.13±1.46条/mm²,显著低于PBS组(34.08±2.46条/mm²,P<0.001); 免疫组织化学MPO染色结果显示Gly组中性粒细胞计数为每200倍视野17.50±1.98个,明显少于PBS组(59.56±4.79个,P<0.001)。

结论：Gly能减轻小鼠角膜急性碱烧伤模型中的角膜炎症反应,并抑制角膜新生血管形成,这为临床上有效防治角膜新生血管类疾病提供了一种新的思路。     

翼状胬肉扩大切除联合自体角膜缘干细胞移植术的效果观察

目的：观察翼状胬肉扩大切除联合自体角膜缘干细胞移植的手术效果。

方法：将59例60眼翼状胬肉患者,随机分为A,B两组,A组27例27眼行翼状胬肉单纯切除; B组32例33眼行翼状胬肉扩大切除联合自体角膜缘干细胞移植,比较两组复发率、角膜创面愈合时间、术后泪膜恢复情况。

结果：术后随访15～25(平均18±3.6)mo,A组复发8眼(30%),B组复发2眼(6%),B组术后角膜创面愈合时间较A组缩短,泪膜破裂时间长,两组差异具有统计学意义。

结论：翼状胬肉扩大切除联合自体角膜缘干细胞移植,角膜创面愈合快,术后复发率低,是一种安全、有效的方法。     

High fat diet induced gut dysbiosis alters corneal epithelial injury response in mice

PurposeCommensal microbiome secretes various metabolites that can exert important effects on the host immunity and inflammation and can alter cellular functions. However, little is known regarding the effect of microbiome on corneal immunity and genetic expression. The purpose of this study is to describe the effect of diet-induced gut dysbiosis on corneal immunity and corneal gene expression after wounding.MethodsThis study is approved by the Animal Care and Use of the University of Illinois. Six-week-old female C57BL6 mice were fed on a normal chow diet (ND), isocaloric low-fat control diet (LFD), or a 21% milk high-fat diet (HFD) for six weeks. 2 mm corneal epithelial debridement was performed (n = 10). Fecal samples from mice were used for microbial diversity analysis (n > 3). Immunofluorescence staining of corneal wholemount tissue post-debridement was used to visualize immune cell distribution. RNA Seq was performed on tissue samples from corneas following debridement.ResultsMice fed differing diets had significant alterations in gut microbial diversities. After corneal debridement, HFD mice experienced delayed wound healing in comparison to LFD mice and ND mice groups. However, fecal transplantation led to normalization of wound closure rates. Increased γδTCR staining was observed in the LFD group, and decreased LY6G was observed in HFD group (p < 0.05). Gene Ontology terms of differentially expressed genes included response to external stimulus, cell proliferation, migration, adhesion, defense response and leukocyte migration. Top over-represented pathways included ECM-receptor interaction, Cytokine-cytokine receptor interaction, Focal adhesion and Leukocyte trans-endothelial migration.ConclusionsGut microbial dysbiosis alters corneal immune cell distribution, corneal response to injury, and genes related to epithelial function and corneal immunity.     

重组人表皮生长因子修复角膜异物剔除后角膜创面的分析

目的:探讨重组人表皮生长因子(recombinant human epidermal growth factor,rhEGF)对角膜异物剔除后角膜创面的修复效果。方法:选取2012-01/2013-01来我院就诊的角膜异物患者102例188眼为研究对象,根据随机数字表法分为治疗组和对照组。两组患者均于裂隙灯下剔除角膜异物,术后治疗组患眼应用重组人表皮生长因子衍生物滴眼液联合妥布霉素进行治疗,对照组患眼仅应用妥布霉素治疗。比较两组治疗3d后的疗效。结果:治疗后3d,治疗组的治愈率(93.7%)明显高于对照组治愈率(76.6%),差异具有统计学意义(P<0.05)。治疗组荧光素染色阴性率为90.4%,高于对照组的46.8%,差异具有统计学意义(P<0.01)。结论:rhEGF能够特异性和角膜上皮细胞受体结合,缩短损伤的角膜上皮的创伤愈合时间,加速愈合,是一种治疗外伤性角膜上皮缺损的有效方法。     

共焦显微镜观察真菌性角膜溃疡多层羊膜移植术后的转归

目的 应用共焦显微镜观察真菌性角膜溃疡多层羊膜移植术后不同随访时期的变化,以评价该种手术方法的临床价值及探讨术后羊膜的转归.方法 选取经两性霉素B脂质体联合局部清创的方法控制感染,但上皮长期不愈的真菌性角膜溃疡患者32例(32眼),其中22例行多层羊膜移植术作为本研究的治疗组,10例患者拒绝手术,继续给予抗真菌药物治疗,作为对照组.所有研究对象角膜溃疡深度≤1/2角膜厚度,角膜溃疡直径≤6mm.术前所有患眼经角膜刮片均证实有真菌感染(包括丝状真菌和酵母样菌).对治疗组患者于术后2周、1个月、3个月及6个月行共焦显微镜检查,同时对照组亦行共焦显微镜检查,观察角膜上皮愈合时间,溃疡深层愈合情况,并于术后6个月行患眼UBM检查,测量角膜厚度.结果 治疗组术后1个月溃疡表面完全愈合,其外层羊膜片于术后10～14 d溶解,原病变区角膜创面上皮化,至术后3个月时深层羊膜部分吸收,瘢痕形成;术后6个月患者原病变区角膜半透明,表面光滑.随访期间真菌无复发.共焦显微镜检查示,术后2周角膜袁层的羊膜细胞溶解,被角膜上皮细胞取代,上皮下可见大量树突细胞;随之上皮下树突状细胞逐渐减少,深层羊膜逐渐溶解,但至术后6个月仍可见羊膜基底膜,上皮下神经纤维未见恢复;术后6个月角膜测厚与正常角膜厚度无显著差异.药物治疗对照组可见感染控制后1个月溃疡表面上皮化,但可见上皮点状缺损,上皮下树突状细胞随病程延长未见明显减少;6个月时角膜厚度明显变薄,与正常角膜厚度具有显著差异.结论 多层羊膜移植是治疗轻度和中度真菌性角膜溃疡的一种有效的方法,共焦显微镜为活体状态下观察深层羊膜转归及溃疡愈合提供了有力的手段.     

软性角膜接触镜对复发性翼状胬肉术后患者舒适度的影响

目的：评价软性角膜接触镜对复发性翼状胬肉术后患者舒适度的作用。

方法：复发性翼状胬肉切除术后患者98例100眼,采用上部自体带角膜缘干细胞的结膜植片,转位并覆盖手术区巩膜表面,50眼术后角膜加用角膜绷带镜5d(试验组A组),另外50眼常规包盖(对照组B组),观察术后角膜创面愈合时间以及各种症状和体征的情况。

结果：胬肉术后1d,A组角膜刺激症状和体征与B组相比,差异有统计学意义(P<0.05); 术后1d,两组患者角膜上皮愈合情况,差异有统计学意义(P<0.05),A组角膜上皮愈合更快。

结论：角膜接触镜可明显改善患者翼状胬肉术后早期的角膜刺激症状,促进角膜创面愈合,减轻患者术后的不适感。     

Effects of amniotic membrane on epithelial wound healing and stromal remodelling after excimer laser keratectomy in rabbit cornea

AIMS: To investigate if the amniotic membrane (AM) promotes epithelial migration while inhibiting stromal remodelling associated with corneal haze after excimer laser keratectomy. METHODS: A wound 150 microm in depth and 6.0 mm in diameter was produced in 40 rabbits using an excimer laser. One eye was randomly chosen to be covered by the AM while the other eye served as a control. Epithelial wound healing was evaluated, together with any morphological changes of the anterior stroma connected with corneal haze. These morphological changes were histopathologically analysed using dichlortriazinyl aminofluorescein (DTAF), Masson trichrome staining, and an image analyser. RESULTS: The AM group had a short latent phase followed by fast epithelial healing (p<0.001) during the early wound healing period and a significant decrease in the inflammatory response, together with a smaller change in the number of keratocytes than the control group. The mean thickness of the regenerated stroma was significantly thinner in the AM group than in the control group at 8 weeks (p<0.0001). The AM group had a more regular architecture of regenerated stromal lamella at 8 weeks and significantly less haze after 4 weeks than the control group (p<0.05). CONCLUSION: Use of the AM as a dressing on a corneal wound created by excimer laser surgery, in which severe haze is expected, may induce rapid epithelial healing with less inflammatory response. The AM may inhibit the irregular synthesis of stromal collagen that is associated with corneal haze.     

低浓度地塞米松对兔角膜上皮细胞的影响

目的: 探讨地塞米松(dexam ethasone,D EX)对培养的兔角膜上皮细胞和角膜上皮伤口愈合的影响。方法: 体外实验原代培养兔角膜上皮细胞, 用四唑盐(M TT)实验和流式细胞仪检测不同浓度 D EX 对兔角膜上皮细胞的作用, 同时通过 H E 、免疫组织化学染色和扫描电镜及临床观察兔角膜上皮细胞刮除后 0.1g/L D EX 用药组和对照组伤口愈合的情况。结果: M TT 检测显示小于 0.1g/L 浓度的 D EX 对兔角膜上皮细胞的生长无明显影响。流式细胞仪检测到 0.1g/L 浓度的 D EX 对兔角膜细胞生长周期无抑制作用。动物实验: 0.1g/L 地塞米松作用后上皮愈合时间显著性缩短, 两组中新生成的上皮细胞均有较强的 PC N A 蛋白的表达。0.1g/L 地塞米松组与对照组相比, 电子显微镜检查角膜基质排列更规则, 炎症反应较轻。结论: 低于 0.1g/L 浓度的地塞米松对培养的兔角膜上皮细胞的生长无抑制作用。0.1g/L 浓度的地塞米松眼液能有效的促进角膜上皮生长并降低炎症反应, 将有利于准分子激光角膜屈光手术后早期角膜伤口愈合。     

Subconjunctival Injection of Regulatory T Cells Potentiates Corneal Healing Via Orchestrating Inflammation and Tissue Repair After Acute Alkali Burn

PurposeThis study aimed to investigate the therapeutic effects and underlying mechanisms of locally delivered regulatory T cells (Tregs) on acute corneal wound healing after alkali burn.MethodsAfter corneal alkali burn, the mice were injected subconjunctivally with regulatory T cells (Tregs) isolated from syngeneic mice. The wound healing process was monitored by clinical manifestation, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). As amphiregulin (Areg) was significantly upregulated, its reparative function in injured corneas was suggested. The hypothesis was further verified via loss- and gain-of-function experiments by administrating the antibody of Areg (anti-Areg) and recombinant Areg (rmAreg).ResultsSubconjunctivally injected Tregs rapidly migrated to injured corneas. The mice treated with Tregs showed prominently reduced corneal opacity, alleviated edema, and faster re-epithelialization compared with the control group. Mechanistically, Treg treatment led to suppressed infiltration of inflammatory cells, along with improved proliferation and inhibited apoptosis of corneal epithelial cells. Tregs expressed upregulated functional markers, including Areg. Expectantly, the levels of Areg in corneas were dramatically higher in the Treg injection group, in line with better corneal restoration. Additional experiments showed that the administration of anti-Areg blunted the reparative effect of Tregs, while exogenous Areg enhanced it. Treg-treated corneas also exhibited less neovascularization and fibrosis at a later reconstruction stage of corneal repair.ConclusionsThe findings showed that the subconjunctival injection of Tregs effectively promoted corneal wound healing by inhibiting excessive inflammation and enhancing epithelial regeneration, with an indispensable reparative role of Areg. Subsequent complications of corneal vascularization and fibrosis were therefore reduced.     

Effect of fortified antibiotic solutions on corneal epithelial wound healing

PURPOSE: To evaluate the influence of fortified antibiotic eyedrops on corneal epithelial wound healing. METHODS: We developed an in vitro epithelial wound-healing model to evaluate the toxicity of antibiotics. An excimer laser was used to create an epithelial defect 1.5 mm in diameter, 70 microm in depth on the central area of porcine cornea. The intact animal globes were maintained in the incubator by a perfusion system. Fortified antibiotics: 10% piperacillin, 5% cefazolin, 0.5% chloramphenicol, 5% vancomycin, 1% amikacin, 2% gentamicin, and 0.1% amphotericin B were applied to the wound in three applications. The wounds were evaluated 24 h after setup with fluorescein stain and a scoring system. RESULTS: The 0.1% amphotericin B and 2% gentamicin disturbed the corneal epithelial healing rate significantly. The remaining antibiotics did not interfere with the epithelial healing rate in our study design. CONCLUSION: Fortified antibiotic eyedrops demonstrated varied degrees of influence on corneal epithelial wound healing. When antibiotic eyedrops are used, both the efficacy and toxicity of the antibiotics should be the major concern. If efficacy is equivalent, less-toxic agents should be given preference.     

角质细胞生长因子促进角膜上皮损伤修复的研究

目的寻找促进角膜上皮损伤修复，治疗持续性角膜上皮缺损的有效方法。方法用３Ｈ－胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入及液体闪烁技术，观察不同浓度的外源性角质细胞生长因子（ｋｅｒａｔｉｎｏｃｙｔｅｇｒｏｗｔｈｆａｃｔｏｒ，ＫＧＦ）对体外培养的人角膜上皮细胞生长的影响，由此推算出有效滴眼液浓度并应用于兔眼。用计算机图形分析系统计算角膜上皮愈合速率；用光镜和电镜评估愈合的质量。结果０．１～１００ｎｇ／ｍｌＫＧＦ有明显促进体外培养的人角膜上皮细胞生长的作用（增长率为２７．６６％～７６．７３％），且呈剂量依赖性（ｒ＝０．９２３３，Ｐ＜０．００１）。１μｇ／ｍｌＫＧＦ滴兔眼，加速了角膜上皮损伤修复（愈合速率，ＫＧＦ组为１．７７±０．２３ｍｍ２／ｈ，与对照组１．４９±０．２４ｍｍ２／ｈ比较，Ｐ＜０．０５）。结论外源性ＫＧＦ对体外培养的人角膜上皮细胞有明显的促生长作用，其滴眼液有加速兔眼角膜上皮创伤修复的作用。