Stem cells are characterized by their self-renewal capacity and the ability to differentiate into any type of mature cell. In addition to normal stem cells, cancerous stem cells have been found to be responsible for the growth and spread of tumors. According to the CSC hypothesis CSC leads to tumorigenesis and tumor growth. Head and neck cancer are the sixth most common cancer and about 300,000 new cases are diagnosed every year. Despite progress in treatment, the 5-year survival rate for this cancer has not improved over the past 30 years. In the hierarchical structure of stem cells present in the human oral epithelium, it is clear that stem cells are the only long-lived inhabitants of the oral epithelium and, therefore, the only cells capable of accumulating the required number of genetic changes for the development of malignancy. The major surface phenotypic markers associated with SC and CSC characteristics include CD133, CD44 and CD24.
Stem cells are undifferentiated cells with varying degrees of potential and plasticity, capable of self-regeneration and multi-purpose differentiation. To date, stem cells have been isolated from three main sources: embryonic stem cells, somatic stem cells (adult stem cells, mesenchymal stem cells) and, more recently, through genetic manipulation of induced pluripotent stem cell (iPSc). While the use of embryonic stem cells is limited to ethical issues, somatic stem cells constitute a more favorable cellular source used in tissue engineering. Postnatal stem cells are isolated from several tissues including the brain, skin, hair follicles, skeletal muscles, bone marrow and dental tissue: Dental pulp stem cells (DPSC) from pulp of permanent teeth; stem cells from exfoliated deciduous teeth (SHED); periodontal ligament stem cells (PDLSCs); apical papilla stem cells (SCAP) and progenitor cells of dental follicles (DFPC). The use of stem cells in the treatment of various diseases of the oral cavity and diseased periodontium is gaining in importance. New treatments that involve tissue engineering, i.e. the use of stem cells for the regeneration of damaged dental tissues give new hope in solving this complex medical problem.
Periodontitis is microbial infection affecting periodontium, the tooth supporting structure and affects >743 million people worldwide. Neural crest-derived stem cells (NCSCs) hold the promise to regenerate the damaged periodontium. These cells have been identified within adult adipose tissue, periodontal ligament, and palatal tissue. Typical enzymatic isolation protocols are expensive, time consuming and often not clinically compliant. Enzyme-free, mechanical dissociation has been suggested as an alternative method of generating cell suspensions required for cell separation and subsequent expansion ex vivo. In our study, samples of rat skeletal muscle tissue were used to appraise the suitability of a novel mincing method of mechanical dissociation against enzymatic digestion with collagenase and dispase. Skeletal muscle is readily available and has been shown to contain NCSC populations. We used a Rigenera-Human Brain Wave® prototype mincer to produce a suspension of skeletal muscle-derived cells modeling NCSCs. We have compared the resulting cell cultures produced via mechanical dissociation and enzymatic dissociation, producing single cell suspensions suitable for Magnetic Cell Sorting (MACs) and Fluorescence-activated cell sorting (FACS). Despite the Countess Automated Cytometry data demonstrating that cell suspensions produced by mechanical dissociation (n=24) contain on average 26.8 times as many viable cells as enzymatic cell suspensions (n=18), enzymatic suspensions produced more successful cell cultures. Spheroids and subsequently adherent cells formed from 4 enzymatic cell suspensions (44.4%) vs. 1 mechanical cell suspension (8.3%). Enzymatic digestion protocols formed spheroids faster and more plentifully than mechanical cell suspensions. Adherent cells and spheroids isolated via both methods appear morphologically similarly to NCSCs from our previous studies.
Every human being, sooner or later, faces tooth loss. The reasons for this can be multiple -a disease of the tooth that begins with a caries lesion, so that, if no adequate treatment is done in time, the process spread to the dental chamber, capturing the pulp and its associated structures, and continued its spread further to the root of the tooth and periapical tissue. Tooth loss can also lead to the disease of soft tissue structures -dental meat initially manifesting inflammation -gingivitis, so that, in case of lack of adequate therapy, the inflammatory process spreads to bone structures and ultimately leads to loss of teeth. Parodontopathy is a disease that in the advanced stage, in stages 3 and 4, leads to oscilation and loss of teeth.
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