Animals ; Cytoplasm ; ultrastructure ; Fibroblasts ; ultrastructure ; Fractures, Bone ; pathology ; Microscopy, Electron ; Osteoblasts ; ultrastructure ; Rabbits ; Wound Healing

Cell patterns in open wound healing are studied by both light and electron microscopic examinations in regards to time sequence, metamorphosis, and functional aspects. Process of the open wound healing clearly exhibited not only time sequence of cllular appearance but also zonation of cells. In the initial stage, until the 3rd day, the neutrophilic polymorphonuclear leukocytes were predominant and particularly concentrated in the scab region. The mononuclear cells were active cells during the 1st to 7th day and were mainly concentrated in the subscab region. The fibroblastic activities started from the 3rd day and became very active during the 5th to the 10th day, and they were concentrated at granulation tissue region. During the process of wound healing, the cellular elements underwent metamorphosis; The neutrophils from normal to swollen and finally degenerating; the mononuclear to macrophages; the fibroblasts from immature to mature actively protein synthesizing cells. The functions of each cellular element can not be determined with certainty. However, the main function of neutrophils in wound healing is likely the formation of front line defense as a part of the scab formation on the surface. And the major function of mononuclear cells is to debride exudates and damaged tissue debris especially at the subscab area and that of the fibroblasts to replace the tissue defect by proliferation and production of fibrous proteins.
Animal ; Epithelium/ultrastructure ; Fibroblasts/ultrastructure ; Leukocytes/ultrastructure ; Rats ; Skin/injuries* ; Skin/pathology ; Wound Healing* ; Wounds, Penetrating/pathology*

INTRODUCTIONAn investigation was carried out to determine the morphological characteristics of fibroblasts in two portions of the vocal fold (VF) mucosa, the macula flava (MF) and Reinke's space (RS), of three different age groups: newborns, adults and geriatrics.

METHODSNormal human VF obtained from autopsy cases were included in this study: four from mature newborns; four from middle-aged adults; and four from geriatric cases. Fibroblasts in RS and MF were investigated by transmission electron microscopy.

RESULTSThe fibroblasts of the MF in both adults and newborns tended to be stellate in shape, with a small nucleus/cytoplasm (N/C) ratio and a well-developed rough endoplasmic reticulum (rER) and Golgi apparatus (GA). Most of the fibroblasts present in RS were oval in newborns and spindle-shaped in adults, with a large N/C ratio and less developed rER and GA. The majority of fibroblasts of the geriatric MF were stellate in shape; while in geriatric RS, the majority of fibroblasts were spindle-shaped with an N/C ratio of 0.5 to 2.0 as in the case of adults. However, the development of rER and GA was less marked in geriatrics than in adults.

CONCLUSIONHistological changes of fibroblasts in the VF mucosa are one of the important causes of the change in voice quality with ageing. Furthermore, geriatric changes in the vocal ligament can be attributed to the activities and the presence of ageing processes in fibroblasts of geriatric VF mucosa.

Adult ; Age Factors ; Aged ; Cell Nucleus ; ultrastructure ; Cytoplasm ; ultrastructure ; Endoplasmic Reticulum ; ultrastructure ; Female ; Fibroblasts ; ultrastructure ; Golgi Apparatus ; ultrastructure ; Humans ; Infant, Newborn ; Laryngeal Mucosa ; ultrastructure ; Male ; Microscopy, Electron, Transmission ; methods ; Vocal Cords ; ultrastructure

OBJECTIVETo observe the changes of L929 cell membrane with atomic force microscope (AFM) after infrasound exposure and to explore the mechanisms of effect of infrasound on cell membrane.

METHODSAfter primary culture, the L929 cells were exposed to infrasound with intensity output of 130 dB and frequency of 16 Hz 2 hours each day for 3 days. The subsequent changes in the membrane of the control cells and the cells exposed to the infrasound were determined by nano-scale scanning with AFM.

RESULTSAfter infrasound exposure, the normal prominence of the membrane became short and the dent became shallow in the 7.5 microm x 7.5 microm and 4.0 microm x 4.0 microm photographs. The prominence appeared as cobblestones. The surface of the membrane became smooth.

CONCLUSIONThe membrane structure of the L929 cells can be changed by infrasound exposure with intensity of 130 dB and frequency of 16 Hz. The change might be one of the characteristics of effect of infrasound on cell membrane.

Animals ; Cell Membrane ; radiation effects ; ultrastructure ; Cells, Cultured ; Fibroblasts ; radiation effects ; ultrastructure ; Mice ; Microscopy, Atomic Force ; Sound ; adverse effects

OBJECTIVETo study the effect of Ti-TiO2 nanotubes with different diameters on the adhesion of fibroblast and osteoblast, and to find which diameter was more favorable for cells' respective adhesion.

METHODSPure titanium sheets were polished and then anodized at different potentials for 1 h with Ti as anode and Pt as cathode. TiO2 nanotubes formed at 1, 5, 10 and 20 V potentials served as experimental groups and polished pure titanium served as control group. Field emission scanning electron microscopy (Fe-SEM) was used to analyze the surface topography. Stained nucleus with Hoechst33342 were used to measure the cell adhesion. The cell shape on the sample surface were analyzed with Fe-SEM.

RESULTSTiO2 nanotube array of different inner diameters from 15 nm to 100 nm were grown on titanium sheets by anodization at potentials from 1 to 20 V. At 30, 60 and 120 min, fibroblast adhesion at nanotubes anodized at 5 V was (141 ± 9), (388 ± 14) and (489 ± 15) respectively, significantly less than any other nanotube surface at the same time (P < 0.01). Nanotubes anodized at 20 V had the least inhibitory effect for fibroblast adhesion with a number of (579 ± 14) at 120 min, and the cell shape was also inhibited. At 30, 60 and 120 min, osteoblast had a significant better adhesion on nanotubes formed at 5 V than it did on any other surface at the same time (P < 0.01), except the control group at 30 min, with the adhesion number of (198 ± 10), (431 ± 10) and (501 ± 10) respectively, and osteoblast had a abundant spread on nanotubes formed at 5 V; while osteoblast adhesion on nanotubes anodized at 20 V was (152 ± 11), (403 ± 9) and (465 ± 12) respectively, less than on any other nanotube surface within the same time (P < 0.05), and the cell shape on the surface changed to be more elongate.

CONCLUSIONSFibroblast adhesion is inhabited more or less on Ti-TiO2 nanotubes of different diameters. Nanotubes formed at 5 V have the most osteoblast adhesion, and inhibit fibroblast adhesion.

Animals ; Cell Adhesion ; Fibroblasts ; cytology ; ultrastructure ; Mice ; Microscopy, Electron, Scanning ; Nanotubes ; chemistry ; Osteoblasts ; cytology ; ultrastructure ; Surface Properties ; Titanium ; chemistry

The objective of this study was to investigate the effect of cytomegalovirus (CMV) infection on actin and microfilament in human embryo fibroblast cells (HF) and to explore the possible relationship with CMV replication. The cell shape was observed by microscopy after the infection of CMV, RT-PCR assay was used to detect the mRNA expression of beta-actin gene, while Westen-blot was used to measure the level of beta-actin protein. CMV immediately early antigen (IE) in HF cells was analyzed by indirect immunofluorescence assay. Microfilament alteration was determined by cytoskeleton fluorescence probe. The results showed that CMV IE was observed in more than 95% of HF cells after infection, primarily located in nucleus. HF cells infected by CMV changed from thin shuttle shape to round and thick ball shape, even detached from wall. Beta-actin got a significant and gradual decreasing of mRNA level in time-dependent manner (P < 0.05). Compared with uninfected group, the expression of beta-actin protein decreased to (74.2 +/- 13.4)% at 96 hours after infection (P < 0.05). In infected HF cells, microfilaments were ruptured, arranged turbulently, as well as cells merged and fluorescence density of microfilament obviously reduced. It is concluded that cytomegalovirus can induce alteration of actin and microfilament, which may be helpful for CMV to infect, replicate and reactivate in host cells.
Actin Cytoskeleton ; metabolism ; ultrastructure ; Actins ; metabolism ; Cell Line ; Cytomegalovirus Infections ; metabolism ; pathology ; Fibroblasts ; pathology ; ultrastructure ; virology ; Humans

The propagation characteristics of virulent duck plague virus (DPV) in duck embryo fibroblast (DEF) were studied by the method of light microscopy observation of DEF cell culture monolayer, electron microscopy observation of infected DEF cell culture, real-time PCR detecting virus propagation. The results demonstrated that on duck embryo fibroblast a number of plaques were formed by DPV 42 h postinfection. Electron microscopy of the ultrathin section of infected duck embryo fibroblasts demonstrated that the nucleic acid of DPV was round in shape with diameter of 35-45 nm and was often in a cluster in the nucleus of DEF. The nucleocapsid of DPV was round in shape with diameter of 90-100 nm and could be observed both in nucleus and cytoplasm of DEF. The mature DPV which had the structures of envelop and tegument was spherical in shape with diameter of 150-300 nm and was located in cytoplasmic vacuoles. DPV penetrated the DEF cell membrane by direct fusion between the viral envelop and the plasma membrane. Progeny viral nucleic acid was produced in the nucleus and the assembled nucleocapsids obtained the structure of tegument in the cytoplasm and obtained the structure of envelop by budding into the cytoplasmic vesicles. The mature DPV particles were released out of the cell through exocytosis of the cytoplasmic vesicles. Detection of DPV by real-time PCR demonstrated that virus in DEF began its obvious propagation 10 h postinfection and virus amount tended to increase until 30 h postinfection. DPV began to be released into the supernatant 22 h postinfection and the DPV amount peaked 50 h postinfection, when the virus content in DEF and supernatant both underwent approximately 10(3) fold increase. DPV mainly existed in the DEF and the virus content in DEF was 10(2)-10(3) fold than the supernatant.
Animals ; Ducks ; embryology ; virology ; Fibroblasts ; virology ; Herpesviridae ; growth & development ; ultrastructure ; Microscopy, Electron ; Polymerase Chain Reaction

Cell apoptosis induced by duck reovirus (DRV)in duck embryo fibroblasts (DEF) was ascertained by light microscope and electron microscopy, DNA Ladder, flow cytometry and fluorescent microscopy. Typical morphological apoptotic features including cell shrinkage and condensation, margination of nuclear chromatin were observed under light microscope and the formation of apoptotic bodies by electron microscopy. DNA ladder was shown by DNA fragment analysis at 24-144h post infection. Flow cytometry showed that the cell apoptosis appeared at 24h and reached it's crest-time at 72-96h, decreased at 144h. Fluorescent microscopy showed that the apoptotic cells which showed green fluorescence appeared at 24h, the number of dead cells which showed red fluorescence increased with the time went by. The results above confirmed that the apoptosis of DEF was successfully induced by DRV.
Animals ; Apoptosis ; physiology ; Cell Nucleus ; ultrastructure ; Cells, Cultured ; DNA Fragmentation ; Ducks ; Embryo, Nonmammalian ; cytology ; Fibroblasts ; cytology ; ultrastructure ; virology ; Flow Cytometry ; Host-Pathogen Interactions ; Microscopy, Electron, Transmission ; Reoviridae ; physiology

This study was undertaken to assess the effect of different surface microtopograph of Ti-Nb-Zr-Sn alloy on the biological behavior and soft tissue integration of rabbits' lacertus fibroblast. The lacertus fibroblasts of Achilles tendon of rabbits were cultured and inoculated on the surface of smooth Ti-flake (control group), and on the surface of sand blast, microgrooves, thread Ti-flakes respectively (3 treatment groups). The cell's growth ratio and attaching status were examined by MTT test and scanning electron microscopy (SEM). The rabbits' lacertus fibroblasts proliferated regularly on the surface of Ti-flakes, and most results of their inter-group comparison showed statistically significant difference (P<0.05), thus indicating the different effects of different surface microtopographs on cell's proliferation; By SEM, the cells were noted to have no obvious directivity on smooth surface; they attached irregularly on the sand blast surface. But in the other two groups, they were oriented regularly along the texture. The cells were almost shuttle-like on the four kinds of surface except for a part of irregular cells on the sand blast surface, and there were plentiful cell-cell junctions. Under high power lens, we found cells with limpid microvilli in the experiment samples. Owing to the cell's fast proliferation and regular attachment in the experiment, we concluded that the regular microgrooved surface was more suitable for meeting the requirement of the soft tissues' physiological function.
Achilles Tendon ; cytology ; Alloys ; chemistry ; Animals ; Cell Proliferation ; Cells, Cultured ; Fibroblasts ; cytology ; ultrastructure ; Rabbits ; Surface Properties ; Titanium ; chemistry

BACKGROUNDThis study characterized the cardiac telocyte (TC) population both in vivo and in vitro, and investigated its telomerase activity related to mitosis.

METHODSUsing transmission electron microscopy and a phase contrast microscope, the typical morphological features of cardiac TCs were observed; by targeting the cell surface proteins CD117 and CD34, CD117 + CD34 + cardiac TCs were sorted via flow cytometry and validated by immunofluorescence based on the primary cell culture. Then the optimized basal nutrient medium for selected population was examined with the cell counting kit 8. Under this conditioned medium, the process of cell division was captured, and the telomerase activity of CD117 + CD34 + cardiac TCs was detected in comparison with bone mesenchymal stem cells (BMSCs), cardiac fibroblasts (CFBs), cardiomyocytes (CMs).

RESULTSCardiac TCs projected characteristic telopodes with thin segments (podomers) in alternation with dilation (podoms). In addition, 64% of the primary cultured cardiac TCs were composed of CD117 + CD34 + cardiac TCs; which was verified by immunofluorescence. In a live cell imaging system, CD117 + CD34 + cardiac TCs were observed to enter into cell division in a short time, followed by an significant invagination forming across the middle of the cell body. Using a real-time quantitative telomeric-repeat amplification assay, the telomerase concentration in CD117 + CD34 + cardiac TCs was obviously lower than in BMSCs and CFBs, and significantly higher than in CMs.

CONCLUSIONSCardiac TCs represent a unique cell population and CD117 + CD34 + cardiac TCs have relative low telomerase activity that differs from BMSCs, CFBs and CMs and thus they might play an important role in maintaining cardiac homeostasis.

Animals ; Antigens, CD34 ; metabolism ; Fibroblasts ; enzymology ; ultrastructure ; Flow Cytometry ; Mesenchymal Stromal Cells ; enzymology ; ultrastructure ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Microscopy, Phase-Contrast ; Myocytes, Cardiac ; enzymology ; ultrastructure ; Proto-Oncogene Proteins c-kit ; metabolism ; Telomerase ; metabolism ; Vimentin ; metabolism