兔自体静脉移植重建泪小管的病理组织学研究
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摘要
研究背景及目的
因泪小管位置表浅,眼睑内侧外伤极易导致泪小管损伤,临床上多见下泪小管损伤,严重外伤常引起泪小管全程损伤,一期手术若不能及时实现泪道再通,延期修复问题较多,成功率低,极易导致患者终生流泪,影响其视力及生活。目前治疗泪小管全程损伤的标准方法有结膜泪囊吻合术和泪道义管置入术。但因结膜泪囊吻合术改变了泪囊生理位置,易使泪囊肌受损,失去正常虹吸作用,术后泪道虽然通畅,但泪溢仍然存在;而泪道义管置入术易因义管移位或肉芽组织增生无法达到预期效果。使用自体静脉移植重建泪小管保持了泪道原有的生理结构及功能且取材方便,经济可行。上世纪六十年代起,陆续开始有临床报道采用自体静脉移植修复泪道,但未见系统的相关实验研究。
本实验研究在泪小管全程损伤泪囊尚存在的情况下,使用自体静脉结膜囊-泪囊移植重建泪小管,观察移植静脉能否存活或其改建情况,为临床应用提供实验资料。
材料与方法
1动物与分组
成年健康日本大耳白兔39只,体重2.0-2.5kg,雌雄兼用,无眼部疾患及头面部异常。以抽签法随机抽取3只设为正常对照组(6眼),剩余36只兔(72眼)左右眼以随机分为睑结膜-泪囊组及泪湖-泪囊组(每组36眼),以术后6个观察时间点为标准将2个实验组分别分为6组,每组12眼。
2手术方法
将兔全麻后取其耳缘静脉剪裁修边后取8-10mm套用于泪道专用硅胶管上备用,由泪点入路切开泪囊并以巩膜穿刺刀于睑结膜或泪湖向泪囊做一通道,将取下的耳缘静脉置入通道内并将血管两端分别与睑结膜/泪湖及泪囊壁吻合,结膜端的硅胶管固定于睑缘,泪囊端硅胶管插入鼻泪管,缝合泪囊切开处。正常对照组不进行手术处理。
3观察项目和时间点
于术后1、3、5、7、14、28天观察实验组泪道通畅情况;术眼一般情况;移植血管病理及形态学特征;免疫组织化学染色分层计数明确上皮层及上皮下层细胞增殖情况。正常对照组观察项目同上,数据作为正常对照使用。
4统计学分析
使用SPSS11.0软件进行统计学分析。计量数据均采用(?)±s表示,睑结膜-泪囊组与泪湖-泪囊组PCNA阳性细胞指数比较采用配对t检验;睑结膜-泪囊组、泪湖-泪囊组分别与正常对照组PCNA阳性细胞指数比较采用独立样本t检验;睑结膜-泪囊组、泪湖-泪囊组、正常对照组荧光素排泄试验阳性时间比较采用单因素方差分析。以α=0.05作为检验水准。
结果
1结膜囊-泪囊静脉移植重建泪小管术后泪道通畅情况
泪湖-泪囊组于术后14天,睑结膜-泪囊组于术后28天,泪道完全恢复导泪功能。术后28天睑结膜-泪囊组荧光素排泄实验阳性时间与泪湖-泪囊组及正常对照组比较差异均无统计学意义(P＞0.05)。
2移植后术眼一般情况
术后1天,结膜严重充血水肿,伴有大量分泌物,泪溢明显;术后3天,结膜充血水肿稍好转,分泌物明显减少,泪溢减轻;术后5天,结膜水肿基本消退,分泌物局限于硅胶管周,泪溢明显好转;术后7天,结膜充血仅限于硅胶管周结膜,水肿已消退,部分兔有轻度泪溢;术后14天,硅胶管周结膜轻度充血,外观上已无明显异常表现;术后28天,外观上已视不及异常表现。睑结膜-泪囊组与泪湖-泪囊组表现无差异。
3移植静脉病理及形态学
3.1大体观
术后1天、3天,移植静脉严重充血高度水肿,管腔内可见大量分泌物;术后5天,充血水肿较前明显好转,渗出物减少;术后7天,充血水肿基本消退,渗出物及坏死组织视不及,血管内壁光滑,与周围组织分界已不明显;术后14天,移植静脉与周围组织颜色基本相同,肉眼难以辨认其分界;术后28天,仅能通过缝线判断移植静脉与周围组织分界。两实验组表现无明显差异。
3.2 HE染色观察结果
术后1天、3天移植静脉内皮细胞开始脱落、坏死,内皮下平滑肌细胞退化;术后5天,平滑肌细胞大量崩溃瓦解,成纤维细胞增生,出现大片红染均质无结构区域;术后7天,移植静脉邻近吻合口处可见吻合口处上皮开始向血管移行,上皮下成纤维细胞增生明显;术后14天,移植静脉被两端相邻组织上皮被覆,且新生上皮向血管中段移行,上皮下成纤维细胞增殖活跃;术后28天,移植静脉上皮层大部分被新生上皮覆盖,上皮下可见胶原纤维平行或交错分布,成纤维细胞分布其中。两实验组病理学表现基本相同。
4移植血管细胞增殖情况观察(PCNA免疫组织化学染色观察结果)
术后3天,移植血管阳性细胞主要位于上皮下,仅有极少数阳性细胞位于上皮层内;术后7天上皮细胞PCNA阳性细胞指数明显增加,上皮细胞及成纤维细胞PCNA阳性细胞指数均达到高峰;7天后上皮细胞及成纤维细胞PCNA阳性细胞指数逐渐下降。术后各个观察时间点睑结膜-泪囊组与泪湖-泪囊组上皮细胞及成纤维细胞的PNCA阳性细胞指数无明显差异(P＞0.05);术后28天睑结膜-泪囊组与泪湖-泪囊组上皮细胞及成纤维细胞PCNA阳性细胞指数均高于正常对照组(P＜0.05)。
结论
1.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管移植血管改建形成膜性管腔。
2.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管后泪湖-泪囊组14天,睑结膜-泪囊组28天可完全恢复导泪功能。
3.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管后结膜囊及泪囊上皮移行形成新的泪小管上皮,28天时上皮覆盖仍未完成;血管壁肌层变性退化改建,28天仍处于增殖期。
Objective
Because of the superficial position of the lacrimal canaliculus,lesions of lacrimal canalicular,espesilly the lower canaliculus,are often associated with eyelid trauma. Severe eyelid trauma often leads to full range lesions of lacrimal Canalicular.If the injury of the lacrimal canaliculus can not be repaired in time by surgery,it may lead to scarring and stenosis,which in turn causes tearing,and brings the uncomfortable to patient.We usually use the conjunctivodacryocystostomy or set the holdfast in the lacrimal passages to repair the lacrimal passages at present.But the physiological position of the dacryocyst is changed when have the conjunctivodacryocystostomy, and the dacryocyst lost the syphonage.So after the operation the patient still have the symptom of dacryostegma;The tube always displaced and the lumens often be obstructed by the granulation tissue when the man-made tube was put into the lacrimal passages.All of these influence the effect of this treatment.Reconstructing lacrimal cancanaliculus by the autogenous vein graft keep the physiological structure and function of the lacrimal passages.And autogenous vein graft can be taken conveniently.From 1960s,using autogenous vein graft to reconstruct lacrimal drainage system were continuously reported,but those thesises are always on clinical application,without experimental study,and could not be recognized by eye doctors.
In this experiment,an animal model with reconstructing lacrimal cancanaliculus by the autogenous vein graft was made in rabbit between the conjunctival sac and lacrimal sac when the lacrimal canaliculus injuried extensively.The healing process of the autogenous vein graft was observed,in order to provide some experimental data for clinical application.
Materials and Methods
1 Animal and grouping
39 healthy adult Japanese albino rabbits without eye diseases were divided into 3 groups randomly:normal control group,palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group.6 eyes of 3 rabbits were randomly assigned as the normal control group,anther 36 rabbits(72 eyes) were divided into 2 surgery group(one eye is in palpebral conjunctiva to lacrimal sac group and another is in lacrimal lacus to lacrimal sac group of each rabbit),36 eyes in each group.Then divided these rabbits into 6 groups at 6 time points and there are 12 eyes in each group.
2 Methods of surgery
Take the vein of the rabbit's ear(length of 10-12mm) after the general anaesthesia,and slip it(length of 8-10mm)onto the silicon tube.Cutting the lacrimal sac from the lacrimal punctum,and make a passage from the conjunctival sac to lacrimal sac.Put the vein into this passage and suture the two sides of it.Fix the silicon tube on the conjunctival sac side on the lid-margin and put the other side into the nasolacrimal canal.Close the cut of the lacrimal sac at last.Normal control group was not operated.
3 Observation items and the time points
Observe the postoperative patent information of lacrimal drainage system,the general state of health of the eyes,the morphology of the autogenous vein and record the pathological changes of the grafted vein by immunohistochemistry stain on the 1th, 3th,5th,7th,14th and 28th post-operative day.
Normal control group was observed with the same items.
4 Statistical analysis
Statistical analysis was performed with SPSS11.0 software.The measurement data expressed as x±s.Palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group were compared using Student's t-test;Palpebral conjunctiva to lacrimal sac group,lacrimal lacus to lacrimal sac group and normal control group were compared using one-way analysis of variance.
Results
1 Postoperative patent information of lacrimal drainage system
Lacrimal lacus to lacrimal sac group on the 14th post-operative day and palpebral conjunctiva to lacrimal sac group on the 28th post-operative day,lacrimal passage have totally recovered the function of lacrimal passage,and the interval time of a positive result of fluorescein disappearance test(FDT) has no significant differences among normal control group,lacrimal lacus to lacrimal sac group and palpebral conjunctiva to lacrimal sac group(P＞0.05)
2 General state of health
The conjunctival congestion and edema appears seriously,and much secretion can be seen,the symptom of dacryostegma is obviously on the 1th post-operative day.
The conjunctival congestion and edema are getting better,the secretion decrease and the symptom of dacryostegma is anesis on the 3th post-operative day.
The conjunctival edema is extinct,and the secretion limited around the silicon tube,the the symptom of dacryostegma improved obviously on the 5th post-operative day.
The conjunctival congestion is just appears around the silicon tube,some of the rabbit still have the symptom of dacryostegrna on the 7th post-operative day.
From 14 day to 28 day after the surgery there is no typical appearance.
The rabbits in palpebral conjunctiva to lacrimal sac group is similar to these in acrimal lacus to lacrimal sac group.
3 Healing process of autograft vein after surgery
3.1 General appearance of the autogenouse vein
On the 1th and 3th post-operative day,the vein hyperemia and edema seriously, there are much secretion in the lumen of blood vessel.On the 5th post-operative day, the hyperemia and edema are getting better,and the secretion is decrease.On the 7th post-operative day,the edema of the conjunctiva is extinct and the dividing line between the vein and the tissue around is not obvious.On the 14th post-operative day, the vein is similar to the tissue around and it is hard to tell the difference.On the 28th post-operative day,it just can recognize the vein from the suture line on the two side. And it appears no conspicuous difference in the 2 surgery groups.
3.2 Result of HE stain
On the 1th and 3th post-operative day,the endothelial cells of autograft vein began to fall off,and the smooth muscle cells degenerated.On the 5th post-operative day,the fibroblast hyperplasia,the smooth muscle cell begin to crash.On the 7th post-operative day,the new endothelial cells migrated to the autograft vein,and the collagenoblast cells proliferated in propria lamina.On the 14th post-operative day,the fibroblast cells proliferated actively in propria lamina.On the 28th post-operative day, most part of the intine of the vein has been covered by the new epithelia and there were still many fibroblast in propria lamina.The result of HE stain seems similar in the 2 surgery groups.
3.3 Cell proliferation of autograft vein(Result of PCNA immunohistochemistry)
Most of the positive cell are under the epithelium on the 3th post-operative day, the index number of the PCNA positive cell of the epithelial cell increase obviously, and the index number of the PCNA positive cell of the epithelial cell and fibroblast reached a peak on the 7th post-operative day.After that it decreased gradually.
The index number of the PCNA positive cell of the epithelial cell and fibroblast bewteen palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group have no significant difference at the six time points we observed(P＞0.05). The index number of the PCNA positive cell of the epithelial cell and fibroblast cell of palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group are significantly more than that in normal control group on the 28th post-operative day(P＜0.05).
Conculsions
1.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),the autogenous vein reconstruct and form a new lumens.
2.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),on the 14th post-operative day in palpebral conjunctiva to lacrimal sac group and on the 28th post-operative day in lacrimal lacus to lacrimal sac group lacrimal passage have recovered the function.
3.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),the grafted vein is covered by the epithelia of palpebral conjunctiva and lacrimal sac,and it is not finished on the 28th post-operative day.The smooth muscle cells degenerated and reconstruct,and the vein graft still in the condition of proliferative.
因泪小管位置表浅,眼睑内侧外伤极易导致泪小管损伤,临床上多见下泪小管损伤,严重外伤常引起泪小管全程损伤,一期手术若不能及时实现泪道再通,延期修复问题较多,成功率低,极易导致患者终生流泪,影响其视力及生活。目前治疗泪小管全程损伤的标准方法有结膜泪囊吻合术和泪道义管置入术。但因结膜泪囊吻合术改变了泪囊生理位置,易使泪囊肌受损,失去正常虹吸作用,术后泪道虽然通畅,但泪溢仍然存在;而泪道义管置入术易因义管移位或肉芽组织增生无法达到预期效果。使用自体静脉移植重建泪小管保持了泪道原有的生理结构及功能且取材方便,经济可行。上世纪六十年代起,陆续开始有临床报道采用自体静脉移植修复泪道,但未见系统的相关实验研究。
本实验研究在泪小管全程损伤泪囊尚存在的情况下,使用自体静脉结膜囊-泪囊移植重建泪小管,观察移植静脉能否存活或其改建情况,为临床应用提供实验资料。
材料与方法
1动物与分组
成年健康日本大耳白兔39只,体重2.0-2.5kg,雌雄兼用,无眼部疾患及头面部异常。以抽签法随机抽取3只设为正常对照组(6眼),剩余36只兔(72眼)左右眼以随机分为睑结膜-泪囊组及泪湖-泪囊组(每组36眼),以术后6个观察时间点为标准将2个实验组分别分为6组,每组12眼。
2手术方法
将兔全麻后取其耳缘静脉剪裁修边后取8-10mm套用于泪道专用硅胶管上备用,由泪点入路切开泪囊并以巩膜穿刺刀于睑结膜或泪湖向泪囊做一通道,将取下的耳缘静脉置入通道内并将血管两端分别与睑结膜/泪湖及泪囊壁吻合,结膜端的硅胶管固定于睑缘,泪囊端硅胶管插入鼻泪管,缝合泪囊切开处。正常对照组不进行手术处理。
3观察项目和时间点
于术后1、3、5、7、14、28天观察实验组泪道通畅情况;术眼一般情况;移植血管病理及形态学特征;免疫组织化学染色分层计数明确上皮层及上皮下层细胞增殖情况。正常对照组观察项目同上,数据作为正常对照使用。
4统计学分析
使用SPSS11.0软件进行统计学分析。计量数据均采用(?)±s表示,睑结膜-泪囊组与泪湖-泪囊组PCNA阳性细胞指数比较采用配对t检验;睑结膜-泪囊组、泪湖-泪囊组分别与正常对照组PCNA阳性细胞指数比较采用独立样本t检验;睑结膜-泪囊组、泪湖-泪囊组、正常对照组荧光素排泄试验阳性时间比较采用单因素方差分析。以α=0.05作为检验水准。
结果
1结膜囊-泪囊静脉移植重建泪小管术后泪道通畅情况
泪湖-泪囊组于术后14天,睑结膜-泪囊组于术后28天,泪道完全恢复导泪功能。术后28天睑结膜-泪囊组荧光素排泄实验阳性时间与泪湖-泪囊组及正常对照组比较差异均无统计学意义(P＞0.05)。
2移植后术眼一般情况
术后1天,结膜严重充血水肿,伴有大量分泌物,泪溢明显;术后3天,结膜充血水肿稍好转,分泌物明显减少,泪溢减轻;术后5天,结膜水肿基本消退,分泌物局限于硅胶管周,泪溢明显好转;术后7天,结膜充血仅限于硅胶管周结膜,水肿已消退,部分兔有轻度泪溢;术后14天,硅胶管周结膜轻度充血,外观上已无明显异常表现;术后28天,外观上已视不及异常表现。睑结膜-泪囊组与泪湖-泪囊组表现无差异。
3移植静脉病理及形态学
3.1大体观
术后1天、3天,移植静脉严重充血高度水肿,管腔内可见大量分泌物;术后5天,充血水肿较前明显好转,渗出物减少;术后7天,充血水肿基本消退,渗出物及坏死组织视不及,血管内壁光滑,与周围组织分界已不明显;术后14天,移植静脉与周围组织颜色基本相同,肉眼难以辨认其分界;术后28天,仅能通过缝线判断移植静脉与周围组织分界。两实验组表现无明显差异。
3.2 HE染色观察结果
术后1天、3天移植静脉内皮细胞开始脱落、坏死,内皮下平滑肌细胞退化;术后5天,平滑肌细胞大量崩溃瓦解,成纤维细胞增生,出现大片红染均质无结构区域;术后7天,移植静脉邻近吻合口处可见吻合口处上皮开始向血管移行,上皮下成纤维细胞增生明显;术后14天,移植静脉被两端相邻组织上皮被覆,且新生上皮向血管中段移行,上皮下成纤维细胞增殖活跃;术后28天,移植静脉上皮层大部分被新生上皮覆盖,上皮下可见胶原纤维平行或交错分布,成纤维细胞分布其中。两实验组病理学表现基本相同。
4移植血管细胞增殖情况观察(PCNA免疫组织化学染色观察结果)
术后3天,移植血管阳性细胞主要位于上皮下,仅有极少数阳性细胞位于上皮层内;术后7天上皮细胞PCNA阳性细胞指数明显增加,上皮细胞及成纤维细胞PCNA阳性细胞指数均达到高峰;7天后上皮细胞及成纤维细胞PCNA阳性细胞指数逐渐下降。术后各个观察时间点睑结膜-泪囊组与泪湖-泪囊组上皮细胞及成纤维细胞的PNCA阳性细胞指数无明显差异(P＞0.05);术后28天睑结膜-泪囊组与泪湖-泪囊组上皮细胞及成纤维细胞PCNA阳性细胞指数均高于正常对照组(P＜0.05)。
结论
1.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管移植血管改建形成膜性管腔。
2.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管后泪湖-泪囊组14天,睑结膜-泪囊组28天可完全恢复导泪功能。
3.兔8-10mm长自体静脉结膜囊-泪囊移植重建泪小管后结膜囊及泪囊上皮移行形成新的泪小管上皮,28天时上皮覆盖仍未完成;血管壁肌层变性退化改建,28天仍处于增殖期。
Objective
Because of the superficial position of the lacrimal canaliculus,lesions of lacrimal canalicular,espesilly the lower canaliculus,are often associated with eyelid trauma. Severe eyelid trauma often leads to full range lesions of lacrimal Canalicular.If the injury of the lacrimal canaliculus can not be repaired in time by surgery,it may lead to scarring and stenosis,which in turn causes tearing,and brings the uncomfortable to patient.We usually use the conjunctivodacryocystostomy or set the holdfast in the lacrimal passages to repair the lacrimal passages at present.But the physiological position of the dacryocyst is changed when have the conjunctivodacryocystostomy, and the dacryocyst lost the syphonage.So after the operation the patient still have the symptom of dacryostegma;The tube always displaced and the lumens often be obstructed by the granulation tissue when the man-made tube was put into the lacrimal passages.All of these influence the effect of this treatment.Reconstructing lacrimal cancanaliculus by the autogenous vein graft keep the physiological structure and function of the lacrimal passages.And autogenous vein graft can be taken conveniently.From 1960s,using autogenous vein graft to reconstruct lacrimal drainage system were continuously reported,but those thesises are always on clinical application,without experimental study,and could not be recognized by eye doctors.
In this experiment,an animal model with reconstructing lacrimal cancanaliculus by the autogenous vein graft was made in rabbit between the conjunctival sac and lacrimal sac when the lacrimal canaliculus injuried extensively.The healing process of the autogenous vein graft was observed,in order to provide some experimental data for clinical application.
Materials and Methods
1 Animal and grouping
39 healthy adult Japanese albino rabbits without eye diseases were divided into 3 groups randomly:normal control group,palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group.6 eyes of 3 rabbits were randomly assigned as the normal control group,anther 36 rabbits(72 eyes) were divided into 2 surgery group(one eye is in palpebral conjunctiva to lacrimal sac group and another is in lacrimal lacus to lacrimal sac group of each rabbit),36 eyes in each group.Then divided these rabbits into 6 groups at 6 time points and there are 12 eyes in each group.
2 Methods of surgery
Take the vein of the rabbit's ear(length of 10-12mm) after the general anaesthesia,and slip it(length of 8-10mm)onto the silicon tube.Cutting the lacrimal sac from the lacrimal punctum,and make a passage from the conjunctival sac to lacrimal sac.Put the vein into this passage and suture the two sides of it.Fix the silicon tube on the conjunctival sac side on the lid-margin and put the other side into the nasolacrimal canal.Close the cut of the lacrimal sac at last.Normal control group was not operated.
3 Observation items and the time points
Observe the postoperative patent information of lacrimal drainage system,the general state of health of the eyes,the morphology of the autogenous vein and record the pathological changes of the grafted vein by immunohistochemistry stain on the 1th, 3th,5th,7th,14th and 28th post-operative day.
Normal control group was observed with the same items.
4 Statistical analysis
Statistical analysis was performed with SPSS11.0 software.The measurement data expressed as x±s.Palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group were compared using Student's t-test;Palpebral conjunctiva to lacrimal sac group,lacrimal lacus to lacrimal sac group and normal control group were compared using one-way analysis of variance.
Results
1 Postoperative patent information of lacrimal drainage system
Lacrimal lacus to lacrimal sac group on the 14th post-operative day and palpebral conjunctiva to lacrimal sac group on the 28th post-operative day,lacrimal passage have totally recovered the function of lacrimal passage,and the interval time of a positive result of fluorescein disappearance test(FDT) has no significant differences among normal control group,lacrimal lacus to lacrimal sac group and palpebral conjunctiva to lacrimal sac group(P＞0.05)
2 General state of health
The conjunctival congestion and edema appears seriously,and much secretion can be seen,the symptom of dacryostegma is obviously on the 1th post-operative day.
The conjunctival congestion and edema are getting better,the secretion decrease and the symptom of dacryostegma is anesis on the 3th post-operative day.
The conjunctival edema is extinct,and the secretion limited around the silicon tube,the the symptom of dacryostegma improved obviously on the 5th post-operative day.
The conjunctival congestion is just appears around the silicon tube,some of the rabbit still have the symptom of dacryostegrna on the 7th post-operative day.
From 14 day to 28 day after the surgery there is no typical appearance.
The rabbits in palpebral conjunctiva to lacrimal sac group is similar to these in acrimal lacus to lacrimal sac group.
3 Healing process of autograft vein after surgery
3.1 General appearance of the autogenouse vein
On the 1th and 3th post-operative day,the vein hyperemia and edema seriously, there are much secretion in the lumen of blood vessel.On the 5th post-operative day, the hyperemia and edema are getting better,and the secretion is decrease.On the 7th post-operative day,the edema of the conjunctiva is extinct and the dividing line between the vein and the tissue around is not obvious.On the 14th post-operative day, the vein is similar to the tissue around and it is hard to tell the difference.On the 28th post-operative day,it just can recognize the vein from the suture line on the two side. And it appears no conspicuous difference in the 2 surgery groups.
3.2 Result of HE stain
On the 1th and 3th post-operative day,the endothelial cells of autograft vein began to fall off,and the smooth muscle cells degenerated.On the 5th post-operative day,the fibroblast hyperplasia,the smooth muscle cell begin to crash.On the 7th post-operative day,the new endothelial cells migrated to the autograft vein,and the collagenoblast cells proliferated in propria lamina.On the 14th post-operative day,the fibroblast cells proliferated actively in propria lamina.On the 28th post-operative day, most part of the intine of the vein has been covered by the new epithelia and there were still many fibroblast in propria lamina.The result of HE stain seems similar in the 2 surgery groups.
3.3 Cell proliferation of autograft vein(Result of PCNA immunohistochemistry)
Most of the positive cell are under the epithelium on the 3th post-operative day, the index number of the PCNA positive cell of the epithelial cell increase obviously, and the index number of the PCNA positive cell of the epithelial cell and fibroblast reached a peak on the 7th post-operative day.After that it decreased gradually.
The index number of the PCNA positive cell of the epithelial cell and fibroblast bewteen palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group have no significant difference at the six time points we observed(P＞0.05). The index number of the PCNA positive cell of the epithelial cell and fibroblast cell of palpebral conjunctiva to lacrimal sac group and lacrimal lacus to lacrimal sac group are significantly more than that in normal control group on the 28th post-operative day(P＜0.05).
Conculsions
1.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),the autogenous vein reconstruct and form a new lumens.
2.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),on the 14th post-operative day in palpebral conjunctiva to lacrimal sac group and on the 28th post-operative day in lacrimal lacus to lacrimal sac group lacrimal passage have recovered the function.
3.Reconstructing lacrimal canaliculus by autogenous vein graft(8-10mm long),the grafted vein is covered by the epithelia of palpebral conjunctiva and lacrimal sac,and it is not finished on the 28th post-operative day.The smooth muscle cells degenerated and reconstruct,and the vein graft still in the condition of proliferative.
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