豚鼠喉粘膜创伤愈合过程的电镜观察

为进一步了解喉粘膜创后超微结构变化的特点，用微型锉对１６只豚鼠喉粘膜造成伤模型，主要以透电镜观察其愈合过程。发现：喉粘膜上皮擦伤缺损的修复始于创缘基底细胞，２４小时后再生上皮开始迁延、移行，胞浆内见少量分布的网状和／或窄幅束状张力丝；５日后覆盖创面的幼稚细胞活跃增殖，胞浆内弥漫分布网状和束状张力丝；１０日后束状张力丝成密集状，介增殖停止代之分化；２１日创面皮分化成熟至正常。再生上皮细胞的增殖能力和     

鼠喉粘膜创伤愈合过程的电镜观察

用刀锉擦伤鼠的声门部粘膜,对其创伤后的愈合过程,用扫描及透射电镜进行了观察。结果发现,创伤部缺损粘膜的上皮修复,是由缺损边缘的基底细胞开始向缺损部移行生长。它们先移行,再增殖,随后分化。经过21天完成再生修复过程。但再生的上皮呈过度增生的倾向。粘膜擦伤后的10日内,有增殖能力的细胞不断增加。再生的上皮细胞内可见到分化良好的张力丝。并出现细胞分化紊乱的有丝分裂活动,犹如在癌细胞中所见的情形。在创伤后     

上颌窦成形术后与传统根治术后粘膜修复和再生的实验观察

本文报告了在犬施行上颌窦根治术和成形术５周后，窦腔粘膜重新上皮化的变化。观察发现：上颌窦成形术后粘膜纤毛系统再生比传统的柯一陆式根治术组更完全。上颌窦根治术后的再生粘膜纤毛细胞和杯状细胞明显减少，并可见部分纤维化和息肉样变。对比实验结果表明，上颌窦成形术行中鼻道窦造口代替下鼻道开窗；窦前壁环钻术代替凿开术，为术后粘膜修复与再生提供了解剖生理环境。     

喉癌花生凝集素受体的研究

应用ＡＢＣ免疫组化技术观察了１７例喉癌、癌旁及正常喉粘膜上皮内花生凝集素（ＰＮＡ）受体分布，结果表明癌细胞内ＰＮＡ呈强阳性表达，毗邻正常喉粘膜上皮细胞未见强阳性表达；在不同的喉癌组织学分级之间，ＰＮＡ的表达呈不同程度，其差异具有显著性（Ｐ＜０．００５）。提示ＰＮＡ受体可能是喉癌的一种重要标志，其分布与喉癌的分化、增殖有关，为喉癌形态学早期诊断和鉴别诊断提供了一种辅助手段。     

喉癌花生凝集素受体的研究

应用ＡＢＣ免疫组化技术观察了１７例喉癌、癌旁及正常喉粘膜上皮内花生凝集素（ＰＮＡ）受体分布，结果表明癌细胞内ＰＮＡ呈强阳表达，毗邻正常喉粘膜上皮细胞未见强阳性表达；在不同的喉癌组织学分级之间，ＰＡＮ的表达呈不同程度，其差异具显著性（Ｐ〈０．００５）。提示ＰＮＡ受体可能是喉癌的一种重要标志，其分布与喉癌的分化，增殖有关，为喉癌形态学早期诊断和鉴别诊断提供了一种辅助手段。     

内镜鼻窦术后合理的局部用药对上颌窦粘膜形态学影响的观察

目的 探讨内镜术后局部用药对上颌窦粘膜的作用。方法 运用光镜和电镜观察了4组上颌窦药物灌注后窦口周围粘膜组织学的变化。结果 治疗前窦口周围粘膜上皮糜烂，脱落，萎缩变薄，表面纤毛大部脱落，细胞间连接松散，线粒体减少，肿胀，治疗后敏感抗生素组上皮细胞修复，排列规则，连接紧密，纤毛再生，长短，粗细均匀，致密，胞内线粒体增多，其内嵴增多；而非敏感抗生素组及生理盐水对照组未见上皮修复。结论 慢性鼻窦炎内镜术后，应采用敏感抗生素抗感染及皮质激素药物抗变态反应。对术后鼻窦粘膜的功能恢复非常重要。     

成人嗅上皮形态学观察

采用光镜和透射电镀对成人嗅粘膜进行形态学观察。嗅粘膜以基底膜为界分为上皮层和固有层；固有层内含有大量的Ｂｏｗｍａｎ腺，其胞浆内含有大量分泌颗粒。上皮层有支持细胞、溴神经原和基底细胞三种成分，前两者胞浆内含有较多的线粒体和内质网。基底细胞接近基底膜，胞浆内细胞器少。在电镜水平成人溴上皮有三种主要的改变：不典型嗅囊，介于嗅上皮和呼吸上皮之间的上皮结构及部分呼吸上皮化生。本观察结果对理解嗅觉生理机制和临     

成人嗅上皮形态学观察

采用光镜和透射电镜对成人嗅粘膜进行形态学观察。嗅粘膜以基底膜为界分为上皮层和固有层；固有层内含有大量的Ｂｏｗｍａｉ腺，其胞浆内含有大量分泌颗粒。上皮层有支持细胞、嗅神经原和基底细胞三种成分。前两者胞浆内含有较多的线粒体和内质网。基底细胞接近基底膜，胞浆内细胞器少，在电镜水平成人嗅上皮有三种主要的改变：不典型嗅囊，介于嗅上皮和呼吸上皮之间的上皮结构及邵分呼吸上皮化生。本观察结果对理解嗅觉生理机制和临床疾病对嗅觉功能的影响将有一定帮助。     

哺乳动物内耳毛细胞损伤再生性修复的研究进展

耳聋是严重影响人类生活质量的顽疾之一，目前尚无根治的方法。各种疾病、噪声、药物等所致感音性耳聋。其实质均在于内耳毛细胞的变性和坏死。鱼类和两栖动物内耳毛细胞因噪声、外伤等因素损伤丢失后，在听觉和前庭器官能自发生成新的毛细胞，从结构和功能上修复受损的感觉上皮。Stone等研究听觉受损的鸟类发现。这些再生的毛细胞可能来源于邻近支持细胞的再生性增殖与分化。通常认为，哺乳动物出生后内耳毛细胞失去再生能力。由毛细胞丢失引发的耳聋是不可治愈的。但也有研究表明。由于外伤和药物作用导致听觉受损可引发哺乳动物内耳前庭器官恢复一定的再生能力嘲。这种再生毛细胞可能有以下几种来源：（1）受损或损伤区周围的支持细胞向毛细胞转化；（2）一些感觉上皮细胞的再分裂；（3）听觉上皮以外的某种干细胞（stemcells）激活，增殖并分化为毛细胞。上述因素中。一般认为支持细胞是新生毛细胞的前体细胞。支持细胞向新生毛细胞转化并替代坏死毛细胞是毛细胞损伤修复的关键因素。     

喉粘膜重建(三维胶原体基质培养)

喉粘膜上皮是由复层鳞状上皮与假复层纤毛上皮组成，复层鳞状上皮是上皮增生、发育异常及癌的好发部位。体外喉粘膜重建可更好地了解喉细胞生物学特性，并对喉病的研究提供模式。传统的单层培养系统对研究不能提供一个满意的环境，因为体内上皮是复层的，与细胞外基质包括间质细胞共存，并暴露于空气中。三维胶原体基质培养系统，将能提供一个与体内喉粘膜相同的环境。体外三维胶原体基质培养喉粘膜重建的操作步骤：①从当地屠宰场获得６个月龄的猪喉，取声门区及声门上区粘膜，经处理，分离获得上皮细胞和成纤维细胞；②用分离的上皮细胞和…     

Effect of fibroblasts on epithelial regeneration on the surface of a bioengineered trachea

OBJECTIVES: Our group applied a tracheal prosthesis, which was composed of polypropylene as the frame and collagenous sponge as the scaffold, to the first human case and had successful results. The objective of this study was to find a way to acquire more rapid re-epithelialization with fibroblasts on this tracheal prosthesis. METHODS: Tracheal epithelial cells, which were isolated from the trachea of rats, were suspended in a collagenous gel. The collagenous gel with fibroblasts was layered on a collagenous sponge. The grafts of this "bioengineered trachea" were implanted into tracheal defects of rats, and the regenerated epithelium on the grafts was histologically examined. RESULTS: Seven days after implantation, stratified squamous epithelium covered almost all of the surface of the gel, and some of the implanted fibroblasts in the gel were lined up just below the epithelium. Fourteen days after implantation, columnar and cuboidal ciliated epithelium covered almost all of the surface of the defects, and the implanted fibroblasts had disappeared. The numbers of regenerated epithelial cells at 14 days after implantation were larger than those of control models without fibroblasts, with statistical significance. CONCLUSIONS: The results suggested that the grafts of bioengineered trachea composed of collagenous sponge and collagenous gel with tracheal fibroblasts accelerated epithelial differentiation and proliferation in vivo.     

Growth of human respiratory epithelium on collagen foil

BACKGROUND: Clinical application of bioartificial tracheal prosthesis must still be regarded as an experimental concept because restoration of a functional respiratory epithelium outlining the prosthesis is still not possible. Tissue engineering as a relatively new biotechnological discipline may offer new methods in expanding differentiated respiratory epithelium in vitro. In this study we compare two different cell and tissue culture procedures for growing human nasal mucosa on commercially available collagen foil. MATERIAL AND METHODS: Harvested specimens of human nasal mucosa (n = 6, 4 x 4 cm) were placed on collagen foil and incubated as tissue cultures for 4, 6 and 8 weeks. A suspension of enzymatically dispersed nasal epithelium seeded on collagen foil (5 x 10(5) cells) served as control. Cell growth and ciliary beat were monitored through an inverted microscope with Hoffman's modulation contrast and video set-up. Histological examination was performed after 4, 6 and 8 weeks. RESULTS: In the tissue cultures, the collagen foil was initially covered with fibroblasts growing from the mucosa specimen before epithelial cells spread out. The epithelial layer showed mostly ciliated cells which developed metachronous ciliary beat after 4 weeks in vitro. Ciliary activity was observed until the end of the experiments in 8 weeks. New cells on the suspension cultures were mesenchymal and did not exhibit any ciliary activity. CONCLUSIONS: Mucosa specimens seem to be more appropriate for tissue engineering of respiratory epithelium than cell suspensions from nasal epithelium. Collagen foil as tissue scaffold initiates epithelial-mesenchymal interaction and may play an important role in epithelial differentiation of new respiratory epithelium.     

Regeneration of respiratory epithelia in the rat after free grafting

Summary Following free grafting of septal mucosa from the rat to the rectus abdominis muscle, the mucosal membrane was found to degenerate into a flat squamous epithelium with loss of the majority of differentiated epithelial cells. Both the goblet cells and ciliated cells regenerated from stem cells, which appear in large numbers 2 days after trasnsplantation. Microvilli were found initially on the surfaces of developing ciliated cells and were replaced 21 days post-grafting by cilia. The number of goblet cells during regeneration at first exceeded that of the ciliated cells, but decreased with the passage of time. After 4 months there were more ciliated cells, with the respiratory epithelium now appearing no different than the initially transplanted nasal mucosa. Correspondence to: F. Bootz     

Experimental surgery of the nose, anteroposterior changes of the mucosa on altering the air-flow

In 20 rabbits one nostril was surgically closed and the mucous membrane studied 4--90 days after the operation. The density of goblet cells was determined anteriorly and posteriorly on whole mounts, epithelial changes on serial sections from 4 different localities on the septum. Anteriorly on the open side damage to the cilia initiated epithelial processes of repair, viz. hyperplasia of basal cells, transformation of these cells into columnar cells, and differentiation into mucous and ciliated cells. On the 16th day the epithelium was again columnar and ciliated. As a consequence of continued trauma new cycles were initiated, but not even after 90 days was there any squamous epithelium. In the middle and posteriorly on the septum no changes were demonstrated, indicating a marked, but gradual decrease in the anteroposterior direction of the influence by the air-flow upon the mucosa. On the closed side there was increased secretory activity and normalization of the epithelium which was changed most anteriorly in normal rabbits.     

Effect of perchloroethylene inhalation on nasal mucosa in mice

An experimental group of 16 male pure-bred mice was exposed to perchloroethylene gas at 300ppm for 6h daily for 5 days. Histopathological study of the nasal mucosa was performed sequentially 1, 2, 4, and 7 days after exposure. Erosion of the olfactory epithelium and dilatation of Bowman's glands were observed from 1 to 7 days after exposure. Atrophy of the olfactory nerves was observed from 4 to 7 days after exposure. At 4 days after exposure, regenerating epithelial cells were observed, indicating that these cells represented the first step of the repair process after exposure. Nonetheless, epithelial degeneration in the nasal mucosa without erosion was observed for 4–7 days after exposure. Such epithelial lesions were more severe in the olfactory mucosa and appeared earlier than in other sites in the respiratory mucosa. The present study revealed that perchloroethylene gas exerted a more potent harmful action on the olfactory mucosa than on the general respiratory mucosa.     

Mucociliary activity of free transplanted respiratory epithelium]

The function of free grafted respiratory epithelium was investigated in inbred rats. In nine of ten animals it could be shown that the respiratory epithelium not only regenerated as an epithelial surface lining but also regained mucociliary activity. The grafted epithelium was able to secrete mucus, which was equal in protein fragmentation to the secretion of normal nasal mucosa. The grafted respiratory epithelium showed normal ciliary function which could be demonstrated by recording the ciliary beat pattern present and by subsequent histological examination. Morphologically, a regular structure of ciliae (9 + 2) was found on the upper pole of the regenerated ciliated cells.     

Epithelial reactions to hydroxyapatite. An in vivo and in vitro study

The intention of this study was to investigate the epithelial reactions to hydroxyapatite ceramic in vivo and in vitro. Shortly after implantation in the rat middle ear, hydroxyapatite was found covered by a mucosal layer. In the early postoperative period the implant was almost completely covered by epithelial cells, which were found to proliferate and also showed migratory activity. After longer intervals the implant was completely covered by epithelium, which was composed predominantly of flat polygonal cells and a relatively small number of ciliated epithelium and goblet cells. All cells showed normal morphology. In vitro experiments showed preservation of the morphology of rat middle-ear mucosa explants with good outgrowth of epithelial cells. In these outgrows, the majority of the cells were flat polygonal, but ciliated epithelium was also seen. No difference was found between the absence and presence of hydroxyapatite. Serially cultured cells displayed normal polygonal morphology, but no ciliated cells were found. Ciliated cells were also absent in control experiments without hydroxyapatite. Growth curves obtained in the absence and presence of hydroxyapatite did not differ significantly from each other.     

Tissue engineering for regeneration of the tracheal epithelium

OBJECTIVES: The slowness of epithelialization on the artificial trachea that has been successfully used in humans is a problem. The purpose of this study was to develop a way to regenerate the epithelium on the surface of this artificial trachea. METHODS: In an in vitro study, isolated rat tracheal epithelial cells were seeded on a collagenous gel that was stratified on a collagenous sponge. Histologic and immunohistochemical examinations were made. In an in vivo study, we transplanted grafts with green fluorescent protein-positive tracheal epithelial cells onto the tracheal defects of normal rats. At 3, 7, 14, and 30 days after the operation, histologic and immunohistochemical examinations were made. RESULTS: In the in vitro study, the 3 layers--the epithelium, gel, and sponge--could be observed. The epithelium expressed cytokeratin 14, cytokeratin 18, and occludin. In the in vivo study, the artificial trachea was covered with epithelium at 3 days after operation, and then the epithelium differentiated from single- or double-stratified squamous epithelium into columnar ciliated epithelium. Green fluorescent protein-positive cells were found 3 days after operation. CONCLUSIONS: We believe that the method used in our experiment is an effective way to regenerate the epithelium on the surface of an artificial trachea. With further experimentation, this method should be suitable for clinical application.     

Regeneration of free grafted respiratory epithelium on severely altered lamina propria.

The lamina propria of nasal mucosa has been thought to play a key role in function and metabolism of respiratory epithelia by allowing exchange of various substances and migratory cells from blood vessels into the epithelium. Here we show that the lamina propria of mucosa transplanted to rat rectus abdominis muscle looses its typical structure and becomes vascularized connective tissue. In the subepithelial space of free grafted respiratory epithelia nasal glands, ducts and nerves degenerate and are no more detectable 3 weeks after transplantation. By contrast the epithelial cell layer regenerates, resulting in a healthy appearing mucosa. Histological and functional controls revealed that regenerated respiratory epithelium lies on a layer of vascularized connective tissue and displays normal cellular differentiation and ciliary function. The present experiments demonstrate that the lamina propria can be replaced by vascularized connective tissue without functional loss of the overlaying epithelium. Thus the results argue against a key role of lamina propria in mucosal function.