ABSTRACT
Statement of the Problem: One major goal of tissue engineering and regenerative medicine is to find an appropriate source of mesenchymal stem cells [MSCs] with higher differentiation ability
Purpose: In this experimental study, the osteogenic and chondrogenic differentiation ability of buccal fat pad derived MSCs [BFP-MSCs] with gingival derived cells [GDCs] were compared
Materials and Method: BFP-MSCs and GDCs were cultured enzymatically and expanded. The expanded cells were analyzed for membrane-associated markers, using flow cytometry. Then the ability of these cells to differentiate into osteocyte and chondrocyte was assessed morphologically and by mRNA expression of collagen I [COLL], BGLA and bone morphogenetic protein 2 [BMP2] using qRT-PCR
Results: Flow cytometry analysis showed that both BFP-MSCs and GDCs expressed the characteristic stem cell markers such as CD73, CD44, and CD90, whereas they did not express hematopoietic markers. Mineralized calcium deposition was observed apparently in BFP-MSCs cultured in osteogenic medium but GDCs showed fewer mineralized nodules. The mRNA expression levels of BGLA and BMP2 showed 7×105 and 733-fold more mRNA expression in BFP-MSCs treated with differentiation media compared to the control group. In chondrogenic differentiation, BFP-MSCs transformed from a spindle to a cuboidal shape while GDCs showed only a slight transformation. In addition, mRNA expression of COLL showed 282-fold higher expression in BFP-MSCs in comparison to the control group. Such significant difference in mRNA expression of BGLA, BMP2, and COLL was not observed in GDCs compared to their corresponding controls
Conclusion: Based on the present results, BFP yields a greater proportion of stem cells compared to gingiva. Therefore, this tissue can be introduced as an easily available source for the treatment of periodontal defects and other maxillofacial injuries
ABSTRACT
Background: salivary gland tumors are among malignancies that have high recurrence rate. Immune responses in salivary gland tumors have not been well elucidated. T helper type 1 [Th1] and Th2 cytokines have been reported to play a role in the outcome of head and neck cancers
Objective: to evaluate the serum levels of interferon gamma [IFN- [gamma]], as the hallmark of Th1 cytokines, and interleukin-4 [IL-4], as the hallmark of Th2 cytokines, in patients with benign and malignant salivary gland tumors in comparison with healthy controls
Methods: fifty patients with benign and 14 patients with malignant salivary gland tumors, as well as 23 healthy individuals were recruited. Serum levels of IFN-[gamma] and IL-4 were measured using ELISA method. Nonparametric tests were used for data analysis
Results: serum levels of IFN-[gamma] and IL-4 were found not to be significantly different in patients compared to the control group [0.68 +/- 0.29 vs. 1.03 +/- 0.57 pg/ml, p=0.58 for IFN-[gamma], 4.57 +/- 1.57 vs. 4.41 +/- 1.31 pg/ml, p=0.28 for IL-4]. IFN-[gamma] and IL-4 serum levels were also not significantly different between patients with benign and malignant salivary gland tumors [p=0.54 and p=0.86, respectively]
Conclusion: the systemic levels of IL-4 and IFN-[gamma] seem not to be associated with salivary gland tumor in our study. Investigation of other cytokines produced by Th1 and Th2 cells are warranted
ABSTRACT
Variations in Cytotoxic T Lymphocyte Antigen-4 [CTLA-4] affect the expression and function of this protein. We aimed to investigate the association of +49 A/G [rs231775], +1822 C/T [rs231779] and +6230 A/G [CT60, rs3087243] genetic variations, as well as the merged haplotypes in CTLA-4 gene with susceptibility to, or progression of head and neck cancer. Eighty patients with confirmed head and neck [HN] cancer [age 54.9 +/- 16.1 years] and 85 healthy age/sexmatched controls [age 56.3 +/- 12.4 years] were enrolled in the study. Genotypes were investigated by the PCR-RFLP method. Arlequin software package was used to check for Hardy-Weinberg equilibration, and to estimate the haplotypes. At position +6230 A/G [CT60], AA genotype, as well as A allele was significantly decreased in patients with HN cancers than controls [18.8% vs. 40.7%, p=0.004; odds ratio=0.34, and 46.3% vs. 61.7, p=0.007; odds ratio=0.53%, respectively]. Nearly the same results were obtained when we compared the subgroup of patients with squamous cell carcinoma of the HN [SCC-HN] with control subjects. The frequencies of genotypes and alleles at other positions were not significantly different between patients and controls, however ACG, GTA and GCA haplotypes emerged from three investigated loci occurred with significantly more frequencies in patients [p<0.0001], while ACA and GTG haplotypes were more frequent among controls [p<0.0001]. Significant differences of haplotypes, genotypes and alleles frequencies resisted the Bonferroni correction. Our results suggest that CT60 A allele, as well as ACA and GTG haplotypes in CTLA-4 gene may have protective roles against HN cancer in Iranian population, while ACG, GTA and specially GCA haplotypes may render susceptibility.
ABSTRACT
CCL22/MDC is a CC chemokine with a critical role in regulation of the immune balance in physiological condition. CCL22/CCR-4 ligation has been documented to participate in the migration of regulatory T [Treg] cells and Th2 lymphocytes to the site of breast tumors; circumstances that are known to be associated with poor prognosis. To investigate the association of a single nucleotide polymorphism [SNP] in CCL22 gene; 16C/A [rs4359426; Asp2Ala], with susceptibility to breast cancer in a sample of Iranian population. Methods: 161 patients with pathologically confirmed breast carcinoma [mean age 49.3 +/- 11.5 yrs] and 178 agematched healthy women [mean age: 49.3 +/- 12.9 yrs] were studied. CCL22 genotypes were investigated by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism [PCR-RFLP] method. Data was verified by direct automated sequencing. Arlequin analysis showed no deviation from Hardy-Weinberg equilibrium. The most frequent genotype in both patient and control groups was wild type CC genotype with frequency of 146 out of 161 [90.7%] among patients and 153 out of 178 [86.0%] in control group [p=0.24]. The frequency of CA genotype was 15 [9.3%] and 23 [12.9%] in patients and controls, respectively [p=0.38]. No AA genotype was observed among patients but this genotype was observed with the frequency of 2 out of 178 [1.1%] in control subjects. The minor allele frequency [MAF] was 0.07 in the population. No correlation was found between the investigated genotypes and clinicopathological characteristics of the patients. Conclusively, results of this investigation do not support the association of 16C/A SNP [rs4359426; Asp2Ala] in CCL22 gene with susceptibility to, and progression of, breast cancer in Iranian population
ABSTRACT
Lentivirus-derived vectors are among the most promising viral vectors for gene therapy which is currently available, but their use in clinical practice is limited due to associated risk of insertional mutagenesis. Gene targeting is an ideal method for gene therapy, but it has low efficiency in comparison to viral vector methods. In this study, we are going to design and construct an integrase-minus lentiviral vector. This vector is suitable for transient expression of gene and gene targeting with viral vector. In this experimental study, three missense mutations were induced in the catalytic domain of Integrase gene in the pLP1 plasmid and resulted D64V, D116A and E152G changes in the amino acid sequence through site directed mutagenesis. The pLenti6.2-GW/EmGFP transfer vector, associated with native and mutated packaging mix, was transfected into 293T cell line. In order to titer the lentivirus stock, the viruses were harvested. Finally, the viruses transduced into COS-7 cell line to assess green fluorescent protein [GFP] gene expression by a fluorescence microscopy. Recombinant and wild lentiviruses titer was about 58×10[6] transducing units/ml in COS-7 cell line. The number of GFP-positive cells transduced with native viruses was decreased slightly during two weeks after viral transduction. In contrast, in the case of integrase-minus viruses, a dramatic decrease in the number of GFP positive cells was observed. This study was conducted to overcome the integration of lentiviral genome into a host genome. Nonintegrating lentiviral vectors can be used for transient gene expression and gene targeting if a Target gene cassette is placed in the lentivirus gene structure. This combination method decreases disadvantages of both processes, such as random integration of lentiviruses and low efficiency of gene targeting
Subject(s)
Disease Vectors , Genetic Therapy , Gene Expression , IntegrasesABSTRACT
Myasthenia gravis [MG] is the most common disorder of neuromuscular junction in which autoantibodies develop against nicotinic acetylcholine receptor for unknown reasons. The association of immunomodulator genes with different autoimmune disease has been studied in recent years. The aim of this study was to investigate correlation between a genetic variation in Stromal Cell Derived Factor-1 [SDF1] and susceptibility to MG in an Iranian population. Genotyping of SDF1 at position 801 G/A was performed by Polymerase Chain Reaction-Restriction Length Polymorphism [PCRRFLP] in 87 patients with confirmed myasthenia gravis and 261 normal control subjects. No statistically significant differences were observed in the frequencies of genotypes and alleles between patients and controls [p>0.05]. Furthermore, no significant differences in the genotype distribution were found between the cases with different stages [p>0.05]. Our data suggest that the SDF1 gene polymorphism at position 801 G/A is not associated with myasthenia gravis