Investigation of the biological activity,mechanical properties and wound healing application of a novel scaffold based on lignin–agarose hydrogel and silk fibroin embedded zinc chromite nanoparticles

Given the important aspects of wound healing approaches, in this work, an innovative biocompatible nanobiocomposite scaffold was designed and prepared based on cross-linked lignin–agarose hydrogel, extracted silk fibroin solution, and zinc chromite (ZnCr₂O₄) nanoparticles. Considering the cell viability technique, red blood cell hemolysis in addition to anti-biofilm assays, it was determined that after three days, the toxicity of the cross-linked lignin–agarose/SF/ZnCr₂O₄ nanobiocomposite was less than 13%. Moreover, the small hemolytic effect (1.67%) and high level of prevention in forming a P. aeruginosa biofilm with low OD value (0.18) showed signs of considerable hemocompatibility and antibacterial activity. Besides, according to an in vivo assay study, the wounds of mice treated with the cross-linked lignin–agarose/SF/ZnCr₂O₄ nanobiocomposite scaffold were almost completely healed in five days. Aside from these biological tests, the structural features were evaluated by FT-IR, EDX, FE-SEM, and TG analyses, as well as swelling ratio, rheological, and compressive mechanical study tests. Additionally, it was concluded that adding silk fibroin and ZnCr₂O₄ nanoparticles could enhance the mechanical tensile properties of cross-linked lignin–agarose hydrogel, and also an elastic network was characterized for this designed nanobiocomposite.

Given the important aspects of wound healing approaches, in this work, an innovative biocompatible nanobiocomposite scaffold was designed and prepared based on cross-linked lignin–agarose hydrogel, extracted silk fibroin solution, and zinc chromite (ZnCr₂O₄) nanoparticles.     

Serum Interleukine-6 Concentration,But Not Interleukine-18, is Associated with Head and Neck Squamous Cell Carcinoma Progression

Inflammation has been linked to various steps in tumorigenesis. Interleukin (IL)-6 and IL-18 are two inflammatory cytokines whose serum concentrations are elevated in several types of cancer, including head and neck squamous cell carcinoma (HNSCC) in some studies. This study was designed to analyze the serum concentrations of these cytokines in Iranian HNSCC patients. Serum IL-6 and IL-18 concentrations were assayed by ELISA commercial kits in 65 untreated patients and 20 healthy volunteers. Serum IL-6 concentration was significantly increased in patients compared to healthy individuals (p < 0.000). IL-6 concentration increased as the tumor stage progressed, and a significant difference appeared between stage IV vs. stage I/II/III (p = 0.03) disease. Although serum IL-18 concentration was higher in patients than in healthy individuals, the difference was not statistically significant (p = 0.06). Moreover, there was no association between serum IL-18 concentration and tumor stage (p = 0.47). A significant difference was observed in serum IL-18 concentration according to the gender with higher IL-18 concentration in male patients (p = 0.01). In conclusion, serum concentration of IL-6 might correlate with the stage of tumor progression in Iranian HNSCC patients. Further studies with larger numbers of patients are required to exclude the possible minor correlation of serum IL-18 concentration with tumor stage.     

Ultrastructural changes in rat ovarian cells of fetuses and neonates from diabetic mothers

Maternal diabetes leads to increased blood glucose concentration in the mother and consequently in the fetus, causing various neonatal problems. This study was conducted to evaluate the effect of maternal diabetes on fetal ovarian structure. Forty adult female rats were divided into two equal groups. Diabetes was induced in one group by alloxan. Both groups became pregnant by natural mating. At days 14 to 20 of pregnancy and day 30, day 60, and day 90 after birth, the fetuses and neonates were collected from both groups and the female genital system was isolated from them. The numbers of germ cells were measured using routine histological techniques in fetuses. In the neonatal ovarian samples, various cellular parameters were determined using transmission electron microscopy (TEM) technique. Results revealed a significant decrease in the number of germ cells in the fetal female gonads in the gonads of fetuses from diabetic mothers compared to those of control. Results of TEM revealed some differences in the luteal cells and oocytes of primordial follicles in ovaries of neonates from diabetic mothers compared to that of control. Mitochondrial abnormalities were seen as mitochondrial cristae were destroyed and vacuolation occurred in the mitochondria and the nuclei of luteal cells were darken and condensed. It is generally concluded that maternal hyperglycemia affects fetal female gonads illustrated by a decrease in the oocytes and alteration in the mitochondria and nuclei of oocytes and luteal cells in neonatal ovaries.     

Adenovirus-mediated p53 gene therapy in pediatric soft-tissue sarcoma cell lines: sensitization to cisplatin and doxorubicin

Sarcomas, or tumors of connective tissue, represent roughly 20% of childhood cancers. Although the cure rate for sarcomas in general has significantly improved in the last 10 years, there continue to be subgroups that are difficult to treat. High-grade or metastatic soft-tissue sarcomas and rhabdomyosarcomas (RMS) of the extremities remain therapeutic challenges and their prognosis is often poor. The future of sarcoma therapy will likely include molecular approaches including gene/protein expression profiling and gene-based therapy. Most sarcomas harbor defects in the p53 or pRb pathways. The tumor suppressor p53 is central to regulation of cell growth and tumor suppression and restoring wild-type p53 function in pediatric sarcomas may be of therapeutic benefit. Studies with adenoviral-mediated p53 gene transfer have been conducted in many cancer types including cervical, ovarian, prostatic and head and neck tumors. Studies of this approach, however, remain limited in pediatric cancers, including sarcomas. Using three viral constructs containing cDNA for wild-type p53, mutant p53 (C135S) and lacZ, we studied the effect of adenoviral-mediated gene therapy in four pediatric sarcoma cell lines, RD and Rh4 (RMS), Rh1 (Ewing's sarcoma) and A204 (undifferentiated sarcoma). Using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we have shown a dose-dependent decrease in cell viability 72 h post-treatment that occurs with Ad-wtp53 but not with Ad-mutp53. Cells treated with Ad-wtp53 show upregulation of the p53 downstream targets, p21(CIP1/WAF1) and bax. Growth curves demonstrate suppression of cell growth over a period of 4 days and cells treated with Ad-wtp53 demonstrate a significant increase in sensitivity to the chemotherapeutic agents, cisplatin and doxorubicin. Our results indicate that restoration of wild-type p53 function in pediatric sarcoma cells could provide a basis for novel approaches to treatment of this disease.     

NOD B-cells are insufficient to incite T-cell-mediated anti-islet autoimmunity

Although it is well established that B-cells are required for the development of diabetes in the nonobese diabetic (NOD) mouse, the nature of their role remains unknown. Herein, we investigate the hypothesis that B-cells in this autoimmune background actively disrupt the tolerant state of those T-cells with which they interact. We demonstrate that NOD B-cells express elevated levels of crucial molecules involved in antigen presentation (including CD21/35, major histocompatibility complex class II, and CD40), alterations that invite the possibility of inappropriate T-cell activation. However, when chimeric animals are generated in which all B-cells are NOD-derived, a tolerant state is maintained. These data demonstrate that although B-cells are required for the development of autoimmunity, they are not sufficient to disrupt established tolerance. Moreover, non-B-cell antigen-presenting cells may be the critical actors in the establishment of the tolerant state; this function may be absent in NOD mice as they are characterized by deficient professional antigen-presenting cell function.     

Immunological patterns identifying disease course and evolution in multiple sclerosis patients

Reliable, and easy to measure, immunological markers able to denote disease characteristics in multiple sclerosis (MS) patients are still lacking. We applied a multivariate statistical analysis on results obtained by measuring-by real-time RT-PCR-mRNA levels of 25 immunological relevant molecules in PBMCs from 198 MS patients. The combined measurement of mRNA levels of IL-1beta, TNF-alpha, TGF-beta, CCL20 and CCR3 was able to distinguish MS patients from healthy individuals. CXCR5, CCL5, and CCR3 combined mRNA levels identify primary progressive MS patients while TNF-alpha, IL-10, CXCL10 and CCR3 differentiate relapsing MS patients. Our results indicate that multi-parametric analysis of mRNA levels of immunological relevant molecules in PBMCs may represent a successful strategy for the identification of putative peripheral markers of disease state and disease activity in MS patients.     

Expression of T cell immunoglobulin- and mucin-domain-containing molecules-1 and -3 (TIM-1 and -3) in the rat nervous and immune systems

Expression of T cell immunoglobulin- and mucin-domain-containing molecules (TIMs) can be used as T helper (Th) differentiation markers in the human and mouse. We examined the expression of TIM-1 and -3 mRNAs in rat MBP(63-88)-induced experimental autoimmune encephalomyelitis (EAE). TIM-3 expression was upregulated in the spinal cord during EAE and following antigen restimulation of the encephalitogenic TCRBV8S2+ population. Interestingly, TIM-3 expression was also detected by in situ hybridization in resident cells of the nervous system. TIM-1 was expressed in B cells but not in resident CNS cells and TIM-1 mRNA levels in spinal cord were unchanged throughout the course of EAE. These results support the notion that TIM-3 can also be used as a Th1 differentiation marker in the rat. However, expression of TIM-1 and -3 is not restricted solely to T cells and the presence of TIM-3 in resident CNS cells may indicate a role for this molecule in the interaction between the nervous and immune systems.