Is it safe to utilize in vitro reconstituted human oral epithelium? An oncogenetic pathway study

Abstract

Cell therapy is a therapeutic strategy used to replace or repair damaged tissue. The epithelium transplantation of cultivated keratinocytes has been applied to several modalities of reconstruction, like oral, urethra and ocular surface. Life and death signals work coordinately to ensure cellular quality control and the viability of an organism. The aim of this study is to verify that culture conditions did not induce genetic mutations through the analysis of the key genes: pAKT, Pten, p53 and MDM2 and investigate the presence of the related proteins in human oral keratinocytes obtained by primary culture and in vitro cultivated. Formalin fixed and paraffin embedded tissues from the oral cavity were utilized as control for normal expression of the related markers and two oral squamous cell carcinoma cell lines provided the expression pattern of the proposed markers in the event of cellular transformation. Akt, PTEN, p53 and MDM2 immunohistochemistry and Western-Blotting analyzes were performed. The results showed the expression levels and intracellular localizations of the four proteins evaluated. These analyzes confirmed that the produced in vitro epithelium is bio-compatible for its utilization as reconstruction and reparatory tissue, however further analyses and additional research on other biomarkers should be performed to analyse the long term engraftment of transplantable primary culture of oral keratinocytes and the long term resistance to cellular transformation.

Antifungal activity of Punicalagin–nystatin combinations against Candida albicans

“…All experimental procedures were approved by the Committee for Ethics in Research on Human Beings of Bauru School of Dentistry, University of São Paulo (CAAE, 44951715.6.0000.5417). HPEC were cultured as previously described by Klingbeil, Mathor, Giudice, Yoshito, and Dos Santos Pinto (2012) (Casaroto et al., 2019; Klingbeil et al., 2012). HGF were cultured in Dulbecco's Modified Eagles Medium (DMEM; Gibco Invitrogen) containing 10% fetal bovine serum (FBS; Gibco Invitrogen) and penicillin/streptomycin (100 IU/ml/100 μg/ml; Gibco Invitrogen).…”

The Beneficial Effect ofEquisetum giganteumL. againstCandidaBiofilm Formation: New Approaches to Denture Stomatitis

“…HPEC were cultivated as previously described by Klingbeil et al [ 30 ] and human monocytes were obtained from thirty-milliliter volumes of peripheral blood, which were drawn from apparently healthy five volunteers. After obtaining peripheral blood mononuclear cells by cell separation using Histopaque 1077 gradients, according to the manufacturer's instructions (Sigma-Aldrich Company), monocytes were counted using neutral red (0.02%) and incubated in 96-well culture plates by 2 h at 37°C in 5% CO 2 .…”

E‐cadherin downregulation and Twist overexpression since early stages of oral carcinogenesis

“…Were used in this study two oral cancer cell lines, FaDu (oropharynx squamous cell carcinoma) and SCC‐25 (tongue squamous cell carcinoma), obtained from American Type Culture Collection (Manassas, VA, USA). For comparative purposes, human oral keratinocytes (HOK) were obtained by primary culture as described before . All cells were cultivated in DMEM medium (Dulbecco) supplemented with 10% fetal bovine serum (FBS) (Gibco‐BRL) and 1% penicillin⁄streptomycin (Gibco‐BRL) at 37°C and 5% CO2.…”