去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

去细胞喉支架的制备及喉肌细胞再生的可行性研究

Objective To prepare a deeelhilarized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct recellularized laryngeal muscles. Methods Perfusion decelluarized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day' s adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. Results Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfnsion method shewed better deculluarized effect. More ventages and collagen fibers but no intact cell or anclei were retained in the decellularized martrix. Porosity measured by Image pro plus 6. 0 was 80. 4% ± 3.2% (x ± s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86. 9% ± 1.5% . After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistoehemical examination indicated that sarcomeric-α actin expressed positively in corresponding areas. Conclusions It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.     

犬去细胞喉支架的生物力学研究

Objective To evaluate the biomechanical characteristics of the decellularized laryngeal scaffold. Methods Ten Chinese adult dogs were randomly divided into two groups: perfusion group (n=5) and control group ( n = 5). The acellular larynx scaffold was obtained from dogs through cranial thyroid arteries perfusion with detergents. Comparative examinations were performed by the macroscopic view,histological view ( hematoxylin and eosin stain, Alcian blue stain and Masson stain), scanning electron microscope (SEM) and biomechanical properties between perfusion group and control group. Results Macroscopic view showed that the decellularized laryngeal scaffold appeared pale asphyxia.HE stain indicated that there were little acellular traces of muscle and mucosa. Alcian blue stain, Masson stain and scanning electron microscope(SEM)suggested that there were no obvious changes about lycosaminoglycan and collagen. The compressive modulus of thyroid cartilage was ( 1.06 ±0. 07) MPa ((-x)±s) in experimental groups and (1.15±0.11) MPa in control group , showing no significant difference (t=1.424,P＞0.05),neither in compressive modulus of annular cartilage(1.68±0.11)MPa in experimental groups and (1.67±0. 09)MPa in control group (t = 0.185, P＞0.05). The tensile strength of thyroid cartilage between experimental (5.74±0.88) MPa and control groups (6.18±1.33) MPa did not have the statistical significance(t =0. 627, P ＞0. 05 ). Conclusion These results indicate that perfusion method can construct a perfect biomechanical acellular larynx scaffold which could be a better selection for laryngeal reconstruction with tissue engineering method.     

Reflex control on laryngeal functions--vibration effect of the laryngeal mucosa on recurrent laryngeal nerve reflexes

Electromyographic (EMG) responses of the intrinsic laryngeal muscle have been investigated to clarify reflexogenic laryngeal controls from a viewpoint of its functional significance during phonation. Twenty-five adult cats were anesthetized with intraperitoneal injection of 4ml/kg of a mixture of 10% urethane and 1% alpha-chloralose. Either the internal branch of the superior laryngeal nerve (ISLN) or the recurrent laryngeal nerve (RLN) was carefully dissected and central end of the dissected nerve was electrically stimulated. EMG of the contra-lateral Thyro-Arytenoid muscle (TA muscle) to the stimulation was recorded using a hooked-wire electrode inserted through the laryngeal mucosa. EMG of the TA muscle evoked by the stimulation of the ISLN were analyzed with respect to its latency and discharge pattern inter-collicular brainstem transsected. Together with the stimulation of the RLN, vibratory stimuli were given mainly to the subglottic mucosa as conditioning stimuli. The vibratory frequency was changed from 50Hz to 400Hz step-wisely. Following results were obtained. 1. EMG response of the contra-lateral TA muscle to the stimulation of the ISLN showed two different kinds of latency, approximately 8-10msec, and 40-60msec. 2. After inter-collicular brainstem transsection, evoked response of the latter disappeared. This result indicates that the ISLN-RLN reflex loop consisted of more than two routes, different in the number of synaptic junctions. 3. The vibratory stimuli given to the laryngeal mucosa had facilitatory effect on the reflexive EMG response evoked by the stimulation of the RLN. 4. This facilitatory effect of the vibratory stimuli disappeared after topical anesthesia of the laryngeal mucosa. 5. The facilitatory effect on the reflex responses was partially increased depending on the vibratory frequencies applied. In conclusion, vibratory stimuli to the laryngeal mucosa reflexively modulate the activity of the intrinsic laryngeal muscles.     

Division of the recurrent laryngeal nerve for idiopathic laryngeal spasm

A rare case of idiopathic laryngeal spasm presented itself as sleep apnea in a middle-aged man. A tracheostomy followed by the division of the recurrent laryngeal nerve relieved all of the symptoms.     

Functional laryngeal abduction following reimplantation of the recurrent laryngeal nerves

Functional reinnervation was established in a patient following complete laryngo-tracheal separation with avulsion of both recurrent laryngeal nerves. Following reattachment of the larynx to the trachea, the severed stumps of the recurrent laryngeal nerves were implanted into the laryngeal abductors (the posterior crico-arytenoid muscles). One year later the patient had good abduction and adduction of her vocal cords. The abduction is thought to be a result of reinnervation by the recurrent laryngeal nerves, the adduction due to the action of crico-thyroid muscle whose innervation was undisturbed by the original injury.     

Reflex control of laryngeal functions in the cat: the effect of vibratory stimuli of the laryngeal mucosa on the laryngeal reflex]

To investigate the effect of vibratory stimuli of the subglottic mucosa on the laryngeal reflex, experiments were performed on cats anesthetized with intraperitoneal injection of a mixture of urethane and chloralose. The external branch of the superior laryngeal nerve was cut, while the internal branch of the superior laryngeal nerve (ISLN) was mounted on stimulating electrodes. Electromyograms (EMG) were recorded from the contralateral thyreoarytenoid (TA), posterior cricoarytenoid (PCA), lateral cricoarytenoid (LCA), and cricothyreoid (CT) muscles. When the ISLN was electrically stimulated, the laryngeal reflex was induced. Short latency (early) and long latency (late) responses were observed in TA, PCA, LCA, and CT. Then, vibratory stimuli were applied to the surface of the subglottic mucosa. Vibratory frequencies used in this study were varied stepwise from 100 Hz to 400 Hz, with the amplitude adjusted at 20 microns. Vibratory stimuli had no effect on early responses but did, however, exert a facilitatory effect on late responses of TA and LCA in the transitional phase from inspiration to expiration and on late responses of PCA in the inspiratory phase. After denervation of ISLN, the vibratory effect on late responses disappeared completely. No significant vibratory effect was observed on CT in any respiratory phase. These results suggest that vibratory stimuli applied to the surface of the subglottic mucosa reflexively facilitate the laryngeal reflex and that ISLN afferents and respiratory drive modulate the laryngeal reflex.     

喉内镜手术治疗喉部疾病的临床应用--介绍一种喉部微创外科技术

目的：探讨新的治疗喉部疾病的微创外科技术--喉内镜手术的临床应用价值。方法：使用从德国STORZ公司新引进的国内首套喉内镜手术系统，治疗包括声带小结、声带息肉、喉乳头状瘤、声带囊肿、会厌囊肿、喉部炎性假瘤、喉部血管瘤等55例病人，并随访2-14个月。结果：55例病人，发音恢复正常者51例，发音功能明显好转者4例。术后1个月动态喉镜检查见所有病人声带粘膜光滑，声带活动及闭合良好，粘膜波正常。结论：喉内镜手术视野广、直接、清晰、照明度高，使手术在微创的前提下彻底清除病灶并最大限度地保留功能，特别是可以避免对喉部良性肿瘤、癌前病变、声带微小癌及喉狭窄等多种喉部疾病采用喉裂开术，有很好的临床推广使用价值。     

喉癌局部生长扩散与喉内"屏障作用"

准确判定患者喉癌的临床分期是有计划治疗的关键所在.加深对喉癌局部生长扩散及生物学特性认识有助于治疗前客观准确地评估病变范围,对正确的选择治疗方法,制定治疗计划仍具有显著的现实指导意义.本文就喉癌分型和局部生长扩散、喉内结构对喉局部生长扩散的限制作用等进行综述.