Andrielle Castilho-Fernandes

Human hepatic stellate cell line (LX-2) exhibits characteristics of bone marrow-derived mesenchymal stem cells

The LX-2 cell line has characteristics of hepatic stellate cells (HSCs), which are considered pericytes of the hepatic microcirculatory system. Recent studies have suggested that HSCs might have mesenchymal origin. We have performed an extensive characterization of the LX-2 cells and have compared their features with those of mesenchymal cells. Our data show that LX-2 cells have a phenotype resembling activated HSCs as well as bone marrow-derived mesenchymal stem cells (BM-MSCs). Our immunophenotypic analysis showed that LX-2 cells are positive for activated HSC markers (αSMA, GFAP, nestin and CD271) and classical mesenchymal makers (CD105, CD44, CD29, CD13, CD90, HLA class-I, CD73, CD49e, CD166 and CD146) but negative for the endothelial marker CD31 and endothelial progenitor cell marker CD133 as well as hematopoietic markers (CD45 and CD34). LX-2 cells also express the same transcripts found in immortalized and primary BM-MSCs (vimentin, annexin 5, collagen 1A, NG2 and CD140b), although at different levels. We show that LX-2 cells are capable to differentiate into multilineage mesenchymal cells in vitro and can stimulate new blood vessel formation in vivo. LX-2 cells appear not to possess tumorigenic potential. Thus, the LX-2 cell line behaves as a multipotent cell line with similarity to BM-MSCs. This line should be useful for further studies to elucidate liver regeneration mechanisms and be the foundation for development of hepatic cell-based therapies.

CD44+/CD133+ immunophenotype and matrix metalloproteinase-9: Influence on prognosis in early-stage oral squamous cell carcinoma.

The purpose of this study was to investigate the expression of CD44 and/or CD133 immunophenotypes and the associated effects of matrix metalloproteinase‐9 (MMP‐9) in early‐stage oral squamous cell carcinomas (SCC) to assess their influence on tumor prognosis.

CD441/CD1331 immunophenotype and matrix metalloproteinase-9

The purpose of this study was to investigate the expression of CD44 and/or CD133 immunophenotypes and the associated effects of matrix metalloproteinase‐9 (MMP‐9) in early‐stage oral squamous cell carcinomas (SCC) to assess their influence on tumor prognosis.

Beneficial Role of Low-Intensity Laser Irradiation on Neural β-tubulin III Protein Expression in Human Bone Marrow Multipotent Mesenchymal Stromal Cells

The purpose of the present study was to evaluate the neural protein expression pattern of human multipotent mesenchymal stromal cells (hMSCs) treated with forskolin (free-form/FF). The study investigated forskolin’s capacity to enhance intracellular levels of cyclic adenosine monophosphate (cAMP) by activating adenylate cyclase and probably by inducing neuron-like cells in vitro. In addition, because nanotechnology is a growing field of tissue engineering, we also assessed the action of a new system called the nanostructured-forskolin (NF) to examine the improvement of drug delivery. Afterwards, the cells were submitted to low-level laser irradiation to evaluate possible photobiostimulatory effects. Investigations using the immunofluorescence by confocal microscopy and Western blot methods revealed the expression of the neuronal marker β-tubulin III. Fluorescence intensity quantification analysis using INCell Analyzer System for β-tubulin III was used to examine significant differences. The results showed that after low-level laser irradiation exposure, there was a tendency to increase the β-tubulin III expression in all groups, as expected in the photobiostimulation process. Notably, this process induced for irradiation was more pronounced in irradiated nanoforskolin cells (INF) compared to non-irradiated free-forskolin control cells (NFFC). However, there was also an increase in β-tubulin III protein expression in the groups: irradiated nanocontrol cells (INC) compared to non-irradiated free-forskolin control cells (NFF) and after treatment with non-irradiated free-forskolin (NFF) and non-irradiated nanoforskolin (NNFC). We concluded that the methods using low-level laser irradiation and/or nanoparticles showed an up-regulation of neural-protein expression in hMSCs that could be used to facilitate cellular therapy protocols in the near future.

The Role of Human Papillomavirus in Head and Neck Cancers

Tobacco and alcohol are well-established risk factors for head and neck squamous cell carcinomas (HNSCC), but it can also develop in individuals not exposed to them. However, only a small proportion of tobacco exposed individuals have developed HNSCC, and there is an emerging tumoral population who lack exposure to these mentioned risk factors, suggesting that others factors can play a role in head and neck carcinogenesis. Over the past two decades, the role of high-risk human papilloma virus (HPV) has been studied through several studies worldwide, and data supporting its role as a causative agent in the development and progression of a subset of HNSCC has been controversial, with considerable variability in frequency depending on the population studied, tumor localization, quality of samples and technical resources utilized for HPV detection. As is the case in cervical and anogenital carcinomas, the most frequently detected high-risk HPVs in HNSCC are the 16 and 18 genotypes. The tonsils and oropharynx are the specific sites associated with higher risk of HPV oncogenic transformation, and investigations suggest that HPV infection in these anatomic sites is an independent risk factor for carcinogenesis. The establishment and maintenance of HPV genomes in the squamous epithelium and HPV-related HNSCC cancer is believed to be originated by oncogenic potential of HPV integration into host DNA genome and their ability to manipulate cell cycle regulators, resulting in deregulated expression of oncoproteins such as E6, which promotes degradation of the tumor suppressor protein p53, allowing cells to evade cell cycle checkpoints, and also E7, which binds to retinoblastoma protein (pRb) and could promote the entrance in S1 phase of cell cycle, leading to disruption of normal cell cycle controls. Following cell division, infected cells leave the basal layer, migrate towards the suprabasal regions and begin to differentiate. Increased understanding of cervical pathogenesis has led to confirmation of HPV as an etiological agent for cancers and consequently to the development of preventive vaccines targeting HPV antigens for the control of cervical cancer. The HPV vaccine was developed as a result of the achievement of core technologies able to produce virus-like