Application of muscle basal lamina containing neural stem cells in repair of spinal cord injury in rats / 中华创伤杂志

Objective To observe effect of muscle basal lamina containing neural stem cells (NSCs) in repair of spinal cord injury.Methods Thirty-six SD rats from the same nest were used in the study and spinal cord hemisection models were induced.The animals were classified to blank control group (clearance of the lesion edge only with isotonic saline),NSCs group (transplantation of NSCs to the edge),NSCs + muscle basal lamina group (transplantation of complex of NSCs and muscle basal lamina to the edge) according to random number table,with 12 rats per group.At weeks 4 and 8,survival and migration of the transplanted cells and compatibility of muscle basal lamina with the host were detected.At weeks 2,4,and 8,the hindlimb function was assayed using BBB scoring system.Results NSCs in NSCs + muscle basal lamina group grew at the lesion edge,migrated to both sides of the edge,and integrated with peripheral tissues.Whereas,few NSCs survived at the lesion edge in NSCs group and inflammatory cell infiltration was notable.At week 2,there was no statistical difference of BBB score among the three groups.At weeks 4 and 8,BBB score in NSCs + muscle basal lamina group (7.92 ± 1.00,11.38 ± 1.51) was significantly higher than that in blank control group (3.82 ± 0.75,3.71 ± 0.76) and NSCs group (6.25 ±1.06,8.25 ± 1.83) (P＜0.05).Conclusion Muscle basal lamina orients growth of NSCs along its lumen,facilitates migration of host cells to ground substance within its lumen,and reduces local inflammatory reaction.

Biocompatibility of human umbilical cord mesenchymal stem cells and acellular muscle bioscaffolds / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.25.008

An experimental study on repairing sciatic nerve defects of rats by human umbilical cord derived mesenchymal stem cells and acellular nerve basal lamina tube / 中华显微外科杂志

Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.

Enfermedad autoinmune: perturbación de la conversación entre la célula blanco (epitelial) y la matriz extracelular / Autoimmune disease: impaired cross-talk between epithelial cell and extracellular matrix

Glándulas salivales de pacientes con síndrome de Sjõgren presentan un aumento en la degradación de componentes de la lámina basal (LB, laminina y colágeno IV) y estroma (colágenos I y III y fibronectina). Estos cambios se correlacionan con un desbalance en la expresión y actividad de metaloproteinasas y sus inhibidores titulares (MMP/TIMP) que desorganiza la LB de acinos y ductos. Esta desorganización es concomitante a una sobreexpresión de lamininas -1 y -5 y a la degradación de nidógenos 1 y -2, que tienen como función establecer puentes de conexión entre laminina y colágeno IV. Cambios post-transcripcionales de la integrina alfa 6 beta 4 están correlacionados con una drástica redistribución de beta 4 en acinos con LB desorganizadas. Estos resultados sugieren que alteraciones en la adhesión célula-matriz y en la formación de contactos célula-célula pueden modificar la señalización de la integrina alfa 6 beta 4 induciendo muerte celular cuando hay una severa interrupción de la célula acinar con la LB.

Increased degradation of basal lamina (BL, laminin and type IV collagen) and stroma (type I and III collagens, and fibronectin) proteins have been observed in salivary glands of patients with Sjõgren’s syndrome. Such changes are associated with imbalanced expression and activity of extracellular matrix metalloproteinases and their tissue inhibitors (MMPs/TIMPs), which contribute to disorganization of the parenchyma basal lamina. Disorganization of the basal lamina is paralleled by an overexpression of laminin-1and -5 and the degradation of nidogens 1 and -2: linker proteins that help maintain the integrity of type IV collagen and laminin networks.Additionally, post-transcriptional changes in alpha 6 beta 4 integrin are associated with a dramatic redistribution of beta 4 in acini, particularly where perturbations in BL organization were apparent. These findings are taken to suggest that changes in acinar cell-matrix adhesion and cell-cell contact formation may alter alpha 6beta 4 integrin signaling, triggering cell death only when severe disruption of cell-BL attachment occurs.

Collagen type Ⅲ is an important linking molecule between generated renal tubules and an artificial polyester interstitium Reticulin as a linker between generated tubules and the interstitium / 中国组织工程研究

In the field of regenerative medicine,much consideration has been given to stem/progenitor cells for the future treatment of acute and chronic renal failure.For this strategy to be effective,however,cell biological information about tubule development within the diseased organ is needed.Unresolved cell-biological issues relating to this kind of treatment include①the integration of stem/progenitor cells,②their differentiation into site-specific cell types,and③the spatial formation of new tubules.To better understand the mechanisms related to this technology,renal tubules were generated at the interphase of an artificial interstitium by using advanced culture techniques.Stern/progenitor cells derived from neonatal rabbit kidney were covered with layers of polyester fleece,placed in a perfusion culture container,and superfused for 13 days with fresh and chemically defined Iscove's Modified Dulbeccos Medium(IMDM) containing aldosterone (1×10-7mol/L).The spatial growth of tubules was registered by scanning electron microscopy(SEM) and on whole mounts or cryosections labeled with soybean agglutinin,silver stain and monoclonal antibodies reacting with collagen type Ⅲor laminin γ1.SEM revealed that the generated tubules were completely covered by a basal lamina.The lamina fibroreticularis exhibited numerous fibers connecting the basal aspect of generated tubules with the surrounding polyester fibers.Cryosections labeled with monoclonal antibodies anti-collagen type Ⅲ and silver stain demonstrated the formation of numerous fibers spanning between the basal lamina of generated tubules and neighboring polyester fibers.In matured kidney tubules the samg arrangement of collagen type Ⅲ fibers is observed as that for generated tubules.This work shows that collagen type Ⅲ is a relevant molecular linker between the basal aspect of generated renal tubules and the polyester fibers of the artificial interstitium.

Immunohistochemical Characterization of Human Synovial Bursa Cells by Light and Transmission Electron Microscopy: Where do These Cells Come From? / Caracterización Inmunohistoquímica de las Células de la Bolsa Sinovial Humana Através de Microscopías de Luz y Transmisión: ¿De dónde Provienen esas Células?

In the present study human synovial bursa specimens were examined by light and transmission electron microscopy. For light microscopical investigation the bursa tissue was stained with azan, haematoxylin-eosin and monoclonal antibodies (CD14, CD33, CD36, CD68, laminin). For electron microscopical investigation the bursa specimens were fixated with Karnovsky's solution and 1,5% osmium tetroxide (Os0(4)) in water distilled and contrasted with 5% uranylacetate and embedded in Epon®. For the first time the antigenic phenotype was characterized and conclusions were drawn about the origin of the synovial bursa cells. Histologically the bursa was divided in two distinct layers; the intima, which is formed by a lining layer and a lamina propria, and a subintimal layer. The intima consisted of macrophage like (type I) and fibroblast like cells (type II). According to the immunohistochemical staining and the electron microscopy the type I cell seemed to be a bone marrow derived monocyte and the more frequently seen type II cell was derived from subintimal fibroblasts. The intimal bursa cell frequently interdigitated and usually communicated by their filopodia (indirect cell-cell-communication). Neither tight or gap junctions nor desmosomes could be documented. Although there was no evidence for the existence of a basal lamina, a concentration of extracellular matrix components beyond the bursa cells was observed. In our study there was no accumulation of laminin around the bursal cells, but striking was a vascular bundle of the intima subintima border zone, which was positive for laminin and CD68 and separated the intima from the subintima. In our opinion this histological structure plays an important role in the regeneration of the lining cells and acts like a barrier between bursa and blood.

En el presente estudio se examinaron bolsas sinoviales humanas a través de microscopía de luz y electrónica de transmisión. Para la microscopía de luz, el tejido de las bolsas se tiñó con Azan, H-E y anticuerpos monoclonales (CD14, CD33, CD36, CD68, laminina). Para la microscopía electrónica las bolsas fueron fijadas con solución de Karnovsky y tetróxido de osmio al 1,5% (Os04) en agua destilada y contrastada con acetato de uranilo al 5% y embebido en Epon®. En primera instada, el fenotipo antigénico fue caracterizado, concluyéndose acerca del origen de las células que componen la bolsa sinovial. Histológicamente la bolsa fue dividida en dos capas distintas - la íntima - la cual es formada por una capa lineal y una lámina propia, y, una subintima. La íntima consistió en células parecidas a macrófagos (Tipo I) y células semejantes a fibroblastos (Tipo II). De acuerdo a la tinción inmunohistoquímica y a la microscopía electrónica, las células tipo I parecen provenir de la médula ósea derivada de monocitos y el más frecuente tipo celular II fue derivadado de los fibroblastos de la subintima. Frecuentemente las células de la íntima de la bolsa se interdigitaban y usualmente se comunicaban a través de sus prolongaciones (comunicación célula indirecta-célula). No se observaron ni uniones abiertas, ni cerradas, ni desmosomas. Aunque no hubo evidencia de la existencia de una lámina basal, se observó una concentración de componentes de matriz extracelular más allá de las células de la bolsa. No hubo acumulación de laminina alrededor de estas células, pero destacada era una banda vascular de la zona límite entre íntima y subintima, la cual fue positiva para laminina y CD68 la cual separaba la íntima de la subintima. En nuestra opinión esta estructura histológica juega un importante rol en la regeneración de las células lineales y actúa como una barrera entre la bolsa y la sangre.

THE PATHOLOGICAL CHANGES OF ULTRASTRUCTURE IN BRONCHIAL MUCOSA OF TYPE 2 DIABETIC PATIENTS / 解剖学报

Objective To investigate the pathological changes of ultrastructure of mucosa in various bronchial segments from type 2 diabetic patients. Methods Sixteen cases of type 2 diabetic patients were selected,2-3 pieces of bronchial mucosa and submucosal tissue of the lesion were taken from various bronchi during bronchoscopy and these samples were observed under a transmission electron microscope. Results The basal lamina of bronchial capillary were diffusely thickened and mostly showed onion-skin like change,protein deposited around and mixed with basal menbrance;irregular highly electron dense materials were found to deposite around capillary,capillary lumen became narrow or even collapsed,neutrophilic leucocyte marginated in lumen and adhered with endothelium;protein deposited in the interstitial;endothelial cells and pericytes had dark cell changes.The cistern of rough endoplasmic reticulum dilated and vesicle formed.Conclusion Bronchial mucosa and its surrounding tissues show characteristic pathological changes of diabetes,bronchial is also the target organ of chronic diabetic damage.

An Immuno -Electron Microscopic Study for Perlecan in the Synovium of Degenerative Arthritis of Knee / 대한해부학회지

This study was designed to observe the expression of perlecan in the normal and degenerative arthritic synovial membrane. By using the immunohistochemical staining and immuno -electron microscopical gold labeling techniques, we observed five materials of normal and degenerative arthritic synovia each. The results were as follows. 1. By the immunohistochemical methods, perlecan -positive staining was seen on the 1 ~2 cell layers of the normal synovial membrane. But, a weaker staining compared to that seen in the normal synovial membrane was found in the degenerative arthritic synovial membrane. 2. Under the electron microscopic observation, perlecan was largely distributed in the rough endoplasmic reticulum of the secretory synovial cell, and in the vacuoles of the phagocytic synovial cell on the normal synovium of the human knee joint. It was also found in the extracellular matrix of the synovial membrane. 3. Perlecan -positive cells were also identified on the degenerative arthritic synovium of the human knee joint. However, fewer perlecan was observed here than that found in the normal synovium. In conclusion, perlecan is synthesized by the secretory synovial cells and degraded by the phagocytic synovial cells. And it, known as a major component of the basement membrane, also proven to exist in the extracellular matrix of the synovial membrane having no basement membrane. From the fact that less perlecan was observed in the degenerative arthritis, perlecan is might to play a major role in the degenerative process.

Effect of basic fibroblast growth factor in tissue engineering peripheral nerve bridges / 中华显微外科杂志

0.05). Conclusion The nerve bridge compounded of bFGF heparin, acellular nerve basal lamina tube and Schwann cells can facilitate nerve regeneration. [Key words] Peripheral nerve; Tissue engineer; Acellular nerve basal lamina tube; Basic fibroblast growth factor(bFGF); Schwann cells

Atrial Morphology Related to Secretory Function and Transport of Metabolites : I. Fenestrations of Basal Lamina in Rat Atrial Endocardium / 대한해부학회지

It was well known that atrial myocytes systhesize atrial natriuretic peptide[ANP], and secrete it into the atrial lumen through the atrial endocardium. But the mechanism for regulation of ANP secretion has not been clearly elucidated, because there was little information of the atrial morphology concerning basal lamina. Basal lamina is surmised as one of barriers that control the movement of ANP, a large molecule. This study was attempted to elucidate the morphological characteristics of basal lamina and connective tissue fibers of atrial endocardial layer by scanning electron microscopy. Basal lamina was exposed by removal of the overlying endothelium. This was achieved by using OsO4 maceration, immersion in aqueous boric acid or EDTA treatment. After removal of the endothelial cell, the specimens were exposed to ultrasonic vibration in case of need. The external surface of basal lamina showed a fairly smooth appearance on the whole, although a few irregular folds are often encountered. Fenestrations, 0.1-1 micrometer in diameter, were randomly observed on the basal lamina, and they were circular to oval in shape. Margin of fenestrations was somewhat distinct and some was divided into two parts by linear structures. The structural differences of fenestrations between right and left atria were not found. The fibroreticular lamina under the basal lamina was revealed by removal of the endothelial cells and their basal lamina. This layer was consisted of interwoven fine fibers. These fine fibers were repeatedly divided and fused, forming reticular network. Some fine fibers connected with basal lamina. Some connective tissue fibers below fibroreticular layer were collected into thick bundles running parallel to myocytes. Above results may serve as a basis for the physiological and morphological studies of atrium.

Interaction of Attachment Complex and Anchoring Fibrils following Corneal Culture

This study was performed to elucidate the structural changes of the attachment complex in the cultured cornea. The three normal corneal tissues were used in this study. The ultrastructural changes of the attachment complex were observed by electron microscopy in one corneal tissue which was not cultured and cultured for 6 days and two corneal tissues which were cultured 10 days and 20 days, respectively. The cornea cultured for 6 days showed changes in the electron density of the hemidesmosome. The basal lamina was focally detached from the cytoplasmic wall of the basal epithelial cells in the cornea cultured for 10 days. The anchoring fibrils within the nuded corneal stroma, which was cultured for 20 days, were markedly decreased in number. These findings suggest that the normal basal epithelial cell, hemidesmosomes and basal lamina might be the essential factors to maintain the networks of normal anchoring fibrils in corneal stroma.

Pleomorphic Xanthoastrocytoma: A Case Report

Pleomorphic xanthoastrocytoma is histologically characterized by marked cellular pleomorphism of lipid-laden neoplastic astrocytes and bizarre giant cells showing mitotic figures and high cellularity. Inspite of its ominous-looking microscopic features, howerver, the prognosis is usually favorable. This tumor develops mainly in the supratentorial area of young people and frequently involves the leptomeninges. We experienced a case of pleomorphic xanthoastrocytoma in 18 year-old-male. In addition to the cellular pleomophism, the prominent reticulin fibers surround the individual tumor cells or the tumor cells nests. Immunohistochemical staining and electron microscopy revealed glial fibrillary acidic protein(GFAP) expression and pericytoplasmic basal lamina in the tumor cells.