The in vitro analysis of migration and polarity of blastema cells in the extracellular matrix derived from bovine mesenteric in the presence of fibronectin / 대한해부학회지

Cell migration is an essential process in embryonic development, wound healing, and pathological conditions. Our knowledge of cell migration is often based on the two dimentional evaluation of cell movement, which usually differs from what occurred in vivo. In this study, we investigated cellular migration from blastema tissue toward bovine decellularized mesentery tissue. In this regard, fibronectin (FN) was assessed to confirm cell migration. Therefore, we established a cell migration model using blastema cells migration toward the extracellular matrix derived from bovine mesenteric tissue. A physiochemical decellularization method was utilized based on freeze-thaw cycles and agitation in sodium dodecyl sulfate and Triton X-100 to remove cells from the extracellular matrix (ECM) of bovine mesenteric tissue. These types of matrices were assembled by the rings of blastema tissues originated from the of New Zealand rabbits pinna and cultured in a medium containing FN in different days in vitro, and then they are histologically evaluated, and the expression of the Tenascin C gene is analyzed. By means of tissue staining and after confirmation of the cell removal from mesenteric tissue, polarity, and migration of blastema cells was observed in the interaction site with this matrix. Also, the expression of the Tenascin C gene was assessed on days 15 and 21 following the cell culture process. The results showed that the three dimentional model of cellular migration of blastema cells along with the ECM could be a suitable model for investigating cell behaviors, such as polarity and cell migration in vitro.

Comparative dermatoglyphic study between autistic patients and normal people in Iran

Autism is a neurodevelopmental disorder originating from early childhood; nevertheless, its diagnosis is in older ages. In addition to heredity, environmental factors are also of great significance in the etiology of the disease. Dermatoglyphic patterns, albeit varied, remain stable for a lifetime and yield a large number of patterns upon examination. Studies have shown a significant association between dermatoglyphics and some diseases, especially genetic ones. We compared fingerprints between patients with autism and normal individuals in a Fars population living in Khorasan-Razavi Province, Iran, in 2015. The right and left hand fingerprints of 104 autistic individuals [case group; age range=5-15 y] were collected using a fingerprint scanner. The same process was performed for 102 healthy individuals, in the age range of 6 to 25 years. All dermatoglyphic patterns and ridge counts were determined. The data were analyzed using the Mann-Whitney nonparametric test and binomial distribution. There was a significant difference in the distribution of the dermatoglyphic patterns on the right and left thumbs and the index fingers between the case and control groups [P<0.05]. The patients had a significantly higher count of loops on their right and left thumbs and their index fingers. A significant decrease in ridge counts for the right and left thumbs and the index fingers was observed in the patients compared to the controls. The results suggested that the patterns were associated with the risk of autism. The patterns may be drawn upon as biometric parameters in the screening of children with autism

effect of maternal nicotine on basement membrane collagen IV of brain microvessels changes in neonatal Balb/C mice

Nicotine can pass through placental blood barrier and accumulate in the developing organs of fetus. Also, entering the breast milk, nicotine can have an effect on the neonates. Investigations have showed that collagen IV is one of the most important micro vessels basement membrane components. In this study, the effect of maternal nicotine exposure in pre and postnatal periods on collagen IV in microvessels of neonatal Balb/C mice brain cortex was studied by immunohistochemistry technique. 24 pregnant Balb/C mice were divided in to 4 groups [6 mice in each group]: two experimental and 2 control groups. The mothers in the 1st experimental group were injected 3 mg/kg nicotine intrapritoneally from the 5th day of pregnancy to parturition daily and in 2nd experimental group the same procedure was repeated to the 10th day after parturition [lactation]. The control groups received the same volume of normal saline during the same time. 10 days after delivery, the brain tissues of newborns were isolated. Then, prepared blocks from fixed brain were cut serially for immunohistochemical assay. The findings of the present study indicated that collagen IV reaction in microvessels basement membrane in the first experimental group increased significantly compared to the first control group [p=0.002]. In addition, collagen IV reaction in microvessels basement membrane in the 2nd experimental group increased significantly compared to the 2nd control group [p=0.002]. However, no significant difference was observed between the two experimental groups. These results suggested that maternal nicotine exposure during prenatal period may increase basement membrane collagen IV expression. Also, nicotine increases in maternal breast milk has no effect on basement membrane collagen IV expression

In vitro decellularization of rabbit lung tissue

The bioscaffold can be used in tissue engineering and regenerative medicine. The scaffolds used in tissue engineering must have high porosity to facilitate accelerated angiogenesis for feeding cells and repelling cell waste outside the scaffold. In this experimental study, we attempted to produce lung three-dimensional scaffold and assay its effect on cell penetration and migration. In an experimental study, rabbit lung tissue was decellularized and used as a scaffold for rabbit blastema cells. The scaffolds were studied on the 15[th] day after culturing. Microscopic features revealed high porosity in the lung tissue scaffold. Electron microscopic imaging also showed collagen and elastin were intact, which are important properties in scaffolds designed for tissue engineering. Migration and permeation of blastema cells into the lung tissue scaffold was also observed. Rabbit lung tissue scaffolds have high porosity. Blastema cells successfully migrated toward and permeated the scaffold inside

Three-dimensional scaffold from decellularized human gingiva for cell cultures: glycoconjugates and cell behavior

We studied both the presence of some carbohydrate compounds in a three-dimensional [3D] matrix harvested from human gingiva and the cell behavior in this matrix. In this experimental research, in order to prepare 3D scaffolds, human palatal gingival biopsies were harvested and physically decellularized by freeze-thawing and sodium dodecyl sulfate [SDS]. The scaffolds were placed within the rings of blastema tissues obtained from a pinna rabbit, in vitro. We evaluated the presence of glycoconjugates and cellular behavior according to histological, histochemical and spectrophotometry techniques at one, two and three weeks after culture. One-way analysis of variance [ANOVA] compared the groups. Extracellular matrix [ECM] remained after decellularization of tissue with 1% SDS. Glycoconjugate contents decreased meaningfully at a higher SDS concentration [p<0.0001]. After culture of the ECM scaffold with blastema, we observed increased staining of alcian blue, periodic acid-Schiff [PAS] and toluidine blue in the scaffold and a number of other migrant cells which was caused by cell penetration into the scaffold. Spectrophotometry results showed an increase in glycosaminoglycans [GAGs] of the decellularized scaffolds at three weeks after culture. The present study has shown that a scaffold generated from palatal gingival tissue ECM is a suitable substrate for blastema cell migration and activity. This scaffold may potentially be useful as a biological scaffold in tissue engineering applications

Comparative assessment of nuclear and nucleolar cytochemical parameters of oral epithelial cells in smokers and non-smokers by methyl green-pyronin staining

A strong relationship exists between cigarette smoking and the development of oral squamous cell carcinoma. Smoking can significantly increase cellular proliferation. Nevertheless, there is little reference in literature to the cytological assessment of oral mucosa in this respect. Changes in nuclear and neucleolar cytomorphometric parameters such as diameter, surface, number and color intensity, in cytologic smears which were collected from normal buccai mucosa of 30 cigarette smokers and 30 non smokers, using methyl green-pyronin staining were studied. Our findings attested to smoking as significant inductive factor in cytochemistry as well as morphologic changes. This technique is a valuable tool

[ vitro histological investigation of interactions between rat's mesenchymal stem cells and human gingival matrix]

Extracellular matrix of natural tissues can be used as a scaffold for reconstructing biological tissues and organs. In this study, decellularized human gingival matrix was used as a scaffold for investigating the interactions of rat's bone marrow mesenchymal stem cells with human gingival matrix. To reach this goal, human gingival tissues were decellularized by two detergents sodium dodecyl sulfate [SDS] and Triton X-100. After washing and sterilization procedures, scaffolds were divided into 3 groups. Low density [LD] group was cultivated with 8x104 cells /cm2, high density [HD] group was cultivated with 8x105 cells/cm2, and control [C] group, was maintained in culture medium without any cells. Microscopic sections were prepared from the scaffoldsbefore and after 1, 2 and 4 weeks of culture with mesenchymal cells and were stained with Hematoxylin-Eosin. Repeated measure ANOVA was used to study the cell density alteration significance within the matrixes and post tests of means comparisons were performed within and between the groups. Also, gingival samples before and after decellularization procedure were investigated by scanning electron microscopy. Histological study of decellularized scaffolds revealed that nuclear and cellular components of the tissues were completely removed. Scanning electron microscopy of the scaffolds indicated that collagen fibers of connective tissue remained intact. Study of the scaffolds 1, 2 and 4 weeks after culture, revealed penetration of mesenchymal stem cells in scaffold, migration of cells towards connective tissue's papilla, and moreover epithelium-like structures. Statistical analysis indicated that cell density in HD group was significantly [P<0.05] higher than LD group. Cell density in both LD and HD groups significantly increased at 2nd week and decreased after4 weeks of culture. According to the results, scaffolds prepared from human gingival matrix can be a suitable scaffold for studying In vitro cell behaviors during oral wound healing