A novel cornea tissue scaffold material was prepared with N-vinly pyrrolidome (NVP) and a biodegradable crosslinking agent by radical polymerization, using azoisobutyronitrile (AIBN) as initiator. Water absorption test and contact angle measure were conducted, and the degradation process of material was investigated. The biocompatibility evaluation was carried out by implantation of material in the rabbits, and by cell culture. The water absorption was over 104%, the contact angle was lower than 41degrees, and the degradation speed in vitro kept steady. The results of implantation in the rabbits showed that the material was almost degraded 3 months later and lots of collagen and cornea stroma cells appeared in it,but there was no inflammation around it. The result of epithelial cells culture showed that the cells conglutinated on the material, but no remarkable cytotoxicity was noted.
Animals ; Biocompatible Materials ; chemistry ; Cornea ; cytology ; Epithelium, Corneal ; cytology ; Materials Testing ; Nitriles ; chemistry ; Prostheses and Implants ; Pyrrolidinones ; chemistry ; Rabbits ; Tissue Engineering

The basement membrane (BM) is crucial in regulating the physical and biological activities of esophageal epithelial cells which attach to the underlying BM. In order to simulate the natural construction of BM, we prepared the fibrous scaffolds using biodegradable polylactide (PLA) and silk fibroin (SF) as the materials via electrospinning technology. BM's proteins containing collagen (IV), laminin, entactin and proteoglycan were extracted from porcine esophagus and coated on the eletrospun fibers. Morphology, mechanical strength, biodegradability and cytocompatibility of the coated and uncoated scaffolds were tested and evaluated using scanning electron micrography, mechanical test system, immunofluorescence assay and western blotting with CK14 as the primary antibody. The fibrous scaffold PLA or PLA/SF, generated from the present protocol had good formation and mechanical and biodegradable properties. After coating with BM's proteins, the scaffold could enhance the growth and differentiation of esophageal epithelial cells, which would contribute to remodel and regenerate the tissue engineered epithelium and further contribute to engineer the whole esophagus in future.
Absorbable Implants ; Basement Membrane ; Biocompatible Materials ; chemistry ; Epithelium ; Esophagus ; physiology ; Fibroins ; chemistry ; Humans ; Nanostructures ; chemistry ; Polyesters ; chemistry ; Regeneration ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

PURPOSES: The cell surface metalloproteinase CD10/ neutral endopeptidase 24.11 (NEP) hydrolyzes a variety of peptide substrates and reduces cellular responses to specific peptide hormones. CD10/NEP has been recognized as modulating peptide-mediated proliferation of lung carcinomas and the normal airway epithelium. The purposes of this study are to evaluate the expression of CD10/NEP in human lung cancers, including non- small cell lung carcinoma (NSCLC) and small cell lung carcinoma (SCLC), and to correlate its expression with several clinicopathologic parameters, including proliferative activity. MATERIALS AND METHODS: CD10/NEP expression and proliferative activity were evaluated by immunohisto chemistry in 55 formalin-fixed and paraffin-embedded NSCLC and SCLC specimens, using anti-Human CD10/ NEP and Ki-67 primary antibodies. The correlations between CD10/NEP expression and either Ki-67 proliferative activity or several clinicopathologic parameters were analyzed by chi-square test or Fisher's exact test. RESULTS: Most NSCLC (76%) and SCLC (80%) cases showed loss of CD10/NEP expression in the tumor cells, whereas the bronchial and alveolar epithelia and stromal fibroblasts in the adjacent healthy lung revealed strong expression of CD10/NEP. Its expression was not correlated with proliferative activity or any of the clinicopathologic parameters except for age. Only in terms of topographical expression was CD10/NEP expression found to be inversely correlated with Ki-67 proliferative activity. CONCLUSION: These results suggest that loss of CD10/ NEP expression may be important in the pulmonary carcinogenesis of both NSCLCs and SCLCs, which is topographically related to NSCLC proliferative activity, especially in the squamous cell type.
Antibodies ; Carcinogenesis* ; Chemistry ; Epithelium ; Fibroblasts ; Humans ; Lung ; Lung Neoplasms ; Neprilysin ; Peptide Hormones ; Small Cell Lung Carcinoma

OBJECTIVE: To establish a protein expression profile of human normal colonic epithelia. METHODS: Two-dimensional gel electrophoresis (2-DE) was applied to separate the total proteins of 20 human normal colonic epithelial tissues. The expression proteins in the human normal colonic epithelia were identified by both matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and electrospray ionization tandem mass spectrometry (ESI-Q-TOF), and the biological function and subcellular locations of the identified proteins were analyzed by bioinformatics. RESULTS: A 2-DE reference map of human normal colonic epithelium was established. On the 2-DE map, 1020+/-50 protein spots were detected, 204 protein spots representing 162 non-redundant proteins were identified, and 37 proteins had posttranslational modification. The identified proteins were categorized into several protein groups according to their functions or subcellular locations, whose data were available at our website (http://www.xyproteomics.org). CONCLUSION A protein expression profile of human normal colonic epithelia is established for the first time, which provides useful information for investigating the physiological functions and pathologic process of colonic epithelia.
Adult ; Aged ; Colon ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; Epithelium ; chemistry ; Humans ; Male ; Middle Aged ; Peptide Mapping ; Protein Array Analysis ; Proteins ; analysis ; chemistry ; genetics

OBJECTIVE: To investigate and compare the growth of human fetal retinal pigment epithelial (RPE) cells seeded onto electrospun polyamide nanofibers (EPN) or etched pore polyester (EPP), and, further, to explore their possible use as prosthetic Bruch's membrane. METHODS: Human fetal RPE cells were planted onto the EPN, EPP and plastic (control) substrates in Transwells. The cultures were assessed with respect to cell attachment at 2, 4, 8 hours and proliferation at 1, 4, 8 days after seeding. Growth and morphology of the cells were monitored under the phase contrast microscope, and the phenotype was identified by immunofluorescence staining with antibodies against tight junction protein ZO-1. Strips of single EPP coated with nothing or EPP coated with EPN was differently implanted into the subretinal space of two P21 RCS rats for two weeks and the histologic slides of the retina were assessed. RESULTS: Cultured human fetal RPE cells were attached to either EPN or EPP substrates (with seeding on plastic substrate as control). After 8 h, the numbers of adherent cells in the EPN, EPP and control groups were 1.23*10(5)/cm(2), 1.70*10(5)/cm(2), and 1.64*10(5)/cm(2), respectively. The number of RPE cells attached to EPN was obviously less than that to both EPP and control (P<0.05). On the first day, the proliferation of cells on EPN was less than that of EPP and control (P<0.05); but by the 8th day in culture, the proliferation of cells on EPN had increased and was higher than proliferation on both EPP and control (P<0.05). All of the RPE cells cultured on EPN and EPP substrates were in monolayer, and the EPN-attached cells resembled the inner collagenous layer of Bruch's membrane. Immunofluorescence staining showed that the RPE cells cultured on EPN and EPP substrates adopted a higher expression of ZO-1 than that on the plastic control substrate. Subretinal implantation of either EPP alone or EPP as a carrier for free EPN for 2 weeks in P21RCS rats resulted in an expected encapsulation and loss of photoreceptor layer. No toxicity or other adverse reaction was observed in the vicinity of the transplant. CONCLUSION EPN and EPP could maintain human fetal RPE cell attachment and proliferation. Both EPN and EPP appeared to be grossly tolerance and biocompatible with subretinal implantation. EPN represents an intriguing prospect for prosthetic Bruch's membrane replacement because of its similarity in structure to native Bruch's membrane.
Animals ; Biocompatible Materials ; chemistry ; Bruch Membrane ; Cell Proliferation ; Cells, Cultured ; Fetus ; Humans ; Membranes, Artificial ; Nanofibers ; chemistry ; Polyesters ; chemistry ; Porosity ; Rats ; Retinal Pigment Epithelium ; cytology ; growth & development ; Tissue Engineering

Animals ; Cell Differentiation ; Epithelium ; pathology ; Humans ; Kidney Diseases ; etiology ; Mesoderm ; pathology ; Protein-Serine-Threonine Kinases ; chemistry ; physiology ; Signal Transduction

Amino Acid Sequence ; Animals ; Cochlea ; growth & development ; metabolism ; Epithelium ; metabolism ; Mice ; Molecular Sequence Data ; Voltage-Gated Sodium Channels ; chemistry ; metabolism

OBJECTIVE: To study the relationship between the postmortem interval (PMI) and the changes of DNA content in rabbit's cornea epithelial and endothelium. METHODS: Using the computerized image analysis to measure the DNA content in the cornea epithelial and endothelium of 105 rabbit's at every six hours during 72 hours after death. RESULTS: The degradation rate of DNA in nuclear has an apparent relationship in 72 hours after death of the rabbits. CONCLUSION The determination of the quantity of DNA in cornea nuclear is a new method to estimate the PMI accurately.
Animals ; DNA/analysis* ; Endothelium, Corneal/chemistry* ; Epithelium, Corneal/chemistry* ; Female ; Forensic Medicine ; Male ; Postmortem Changes ; Rabbits ; Signal Processing, Computer-Assisted ; Time Factors

One hundred large bowel carcinomas were studied immunohistochemically with regard to expression of HLA-DR antigen (DR). One or two sections from each tumor including surrounding normal mucosa were examined by a semiquantitative counting system for tumor cells and mucosal and stromal infiltrates of lymphocytes and mononuclear cells (MNCs) with DR expression and the results were applied Chi-square test. The rate of presence of DR positive (DR+) lymphocytes in lymphoid nodules and DR+ lymphocytes/ MNC in the adjacent mucosa and stroma in DR+ carcinoma (50%) was higher (P < 0.01) than in DR- carcinoma (21.9%). Thirty-six carcinomas (36%) were DR+. Three (75%) out of 4 DR+ poorly differentiated carcinomas and six (20%) out of 30 DR+ moderately differentiated carcinomas showed homogeneously strong DR+ expression. There was tendency for poorly differentiated carcinoma to be more homogeneous DR+ expression. According to Dukes' stage, four (80%) out of 5 carcinomas in Dukes' stage D were DR-. An increased infiltration of lymphocytes/MNCs into adjacent mucosa and stroma in large bowel carcinomas is possibly related with DR expression by carcinoma. From the results of this study, we postulated as follows: 1) DR+ tumor cells may act as antigen-presenting cells, 2) They may have an inhibitory effect for distant metastasis, 3) Poorly differentiated carcinoma expressed more DR+ homogeneously.
Adult ; Aged ; Antibodies ; Colorectal Neoplasms/blood/*chemistry/pathology ; Epithelium/chemistry ; Female ; HLA-DR Antigens/*analysis ; Human ; Immunohistochemistry ; Leukocytes, Mononuclear/chemistry ; Lymphocytes/chemistry ; Male ; Middle Age ; Neoplasm Staging ; Support, Non-U.S. Gov't

To study the possibility of using hydroxypropyl chitosan-based blend membranes as carriers of corneal cells in tissue engineering, we prepared three kinds of blend membranes labeled hydroxypropyl chitosan/chondroitin sulfate, hydroxypropyl chitosan/gelatin/chondroitin sulfate and hydroxypropyl chitosan/oxidized hyaluronic acid/chondroitin sulfate. The transparency, water content and ability of protein adsorption of the blend membranes were measured. To evaluate the cytocompatibility of the blend membranes with corneal epithelial cells, rabbit corneal epithelial cells were cultured on the surface of the carrier membranes. The morphological characteristics, cell adhesion, cell proliferation and the activity of lactate dehydrogenase (LDH) in the media were investigated. Three kinds of blend membranes had good optical transmittance, suitable water content and ability of protein adsorption. The results showed that the less injury was made to corneal epithelial cells by the hydroxypropyl chitosan/gelatin/chondroitin sulfate blend membrane than the others. This kind of membrane was favor of the growth and adhesion of corneal epithelial cells. The hydroxypropyl chitosan/gelatin/chondroitin sulfate blend membrane is a promising carrier of corneal cells and can be used in reconstruction of tissue engineered cornea.
Animals ; Biocompatible Materials ; chemistry ; pharmacology ; Cell Culture Techniques ; methods ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chitosan ; chemistry ; Chondroitin Sulfates ; chemistry ; Epithelium, Corneal ; cytology ; Gelatin ; chemistry ; Membranes, Artificial ; Rabbits ; Tissue Engineering ; methods