Background: Cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions. Chondral lesion is a pathology with high prevalence, reaching as much as 63% of general population and 36% among athletes. Despite autologous chondrocyte implantation versatility, it still fails to fully reproduce hyaline articular cartilage characteristics. Mesenchymal stem cells (MSCs) may be isolated from various known tissues, including discarded fragments at arthroscopy such as synovial membrane. Choice of harvesting site is motivated by MSCs' abilities to modulate immunologic and inflammatory response through paracrine communication. Synovial MSCs have a greater proliferation and strong chondrogenic potential than bone and adipose MSCs and a less hypertrophic differentiation than bone MSCs. Good manufacturing practice (GMP) laboratory techniques for human clinical trials are still novel. To our knowledge, there are only two clinical trials in humans published since today.Purpose: Therefore, this work aimed to isolate and characterize synovial MSCs and evaluated their differentiation properties according to GMP standards.Materials and Methods: One-gram tissue sample from three patients of synovia was harvested at the beginning of arthroscopy surgery. MSCs were isolated, expanded, and characterized by flow cytometry.Results: It was possible to isolate and expand MSCs cultures from synovia, characterize MSCs by flow cytometry using proper monoclonal antibodies, and differentiate MSCs by coloring technique after chondrogenic, adipogenic, and osteogenic differentiations. Cartilage treatment may benefit from these tissue engineering protocols since arthroscopic procedures are routinely performed for different purposes in a previous stage and a favorable chondronegic differentiation cell lineage may be collected and stored in a less invasive way.Conclusion: Laboratory protocols established according to presented GMP were able to isolate and characterize MSCs obtained from synovia.Impact StatementArticular cartilage restoration is a desperately needed bridge for patients with symptomatic cartilage lesions and it rises as a socioeconomic issue with a considerable economic burden. Synovial mesenchymal stem cells (MSCs) have a greater proliferation rate and strong chondrogenic potential than bone and adipose MSCs and a less hypertrophic differentiation than bone MSCs. To our knowledge, there are only two human clinical trials with good manufacturing practice laboratory techniques for synovial MSCs harvesting and differentiation. Cartilage treatment may benefit from these tissue engineering protocols since arthroscopic procedures are routinely performed for different purposes in a previous stage.
Background A new trend in the treatment for alveolar clefts in patients with cleft lip and palate involves the use of bone tissue engineering strategies to reduce or eliminate the morbidity associated with autologous bone grafting. The use of mesenchymal stem cells—autologous cells obtained from tissues such as bone marrow and fat—combined with various biomaterials has been proposed as a viable option for use in cleft patients. However, invasive procedures are necessary to obtain the mesenchymal stem cells from these two sources. To eliminate donor site morbidity, noninvasive stem cell sources such as the umbilical cord, orbicularis oris muscle, and deciduous dental pulp have been studied for use in alveolar cleft bone tissue engineering. In this study, we evaluate the osteogenic potential of these various stem cell types. Methods Ten cellular strains obtained from each different source (umbilical cord, orbicularis oris muscle, or deciduous dental pulp) were induced to osteogenic differentiation in vitro, and the bone matrix deposition of each primary culture was quantified. To evaluate whether greater osteogenic potential of the established mesenchymal stem cell strains was associated with an increase in the expression profile of neural crest genes, real-time qPCR was performed on the following genes: SRY-box 9, SRY-box 10, nerve growth factor receptor, transcription factor AP-2 alpha, and paired box 3. Results The mesenchymal stem cells obtained from deciduous dental pulp and orbicularis oris muscle demonstrated increased osteogenic potential with significantly more extracellular bone matrix deposition when compared to primary cultures obtained from the umbilical cord after twenty-one days in culture (p = 0.007 and p = 0.005, respectively). The paired box 3 gene was more highly expressed in the MSCs obtained from deciduous dental pulp and orbicularis oris muscle than in those obtained from the umbilical cord. Conclusion These results suggest that deciduous dental pulp and orbicularis oris muscle stem cells demonstrate superior osteogenic differentiation potential relative to umbilical cord-derived stem cells and that this increased potential is related to their neural crest origins. Based on these observations, and the distinct translational advantage of incorporating stem cells from noninvasive tissue sources into tissue engineering protocols, greater study of these specific cell lines in the setting of alveolar cleft repair is indicated.
Mesenchymal stem cells (MSCs) have been studied as a promising type of stem cell for use in cell therapies because of their ability to regulate the immune response. Although they are classically isolated from the bone marrow, many studies have sought to isolate MSCs from noninvasive sources. The objective of this study was to evaluate how MSCs isolated from the dental pulp of human exfoliated deciduous teeth (SHED) and fragments of the orbicularis oris muscle (OOMDSCs) behave when treated with an inflammatory IFN-γ stimulus, specifically regarding their proliferative, osteogenic, and immunomodulatory potentials. The results demonstrated that the proliferation of SHED and OOMDSCs was inhibited by the addition of IFN-γ to their culture medium and that treatment with IFN-γ at higher concentrations resulted in a greater inhibition of the proliferation of these cells than treatment with IFN-γ at lower concentrations. SHED and OOMDSCs maintained their osteogenic differentiation potential after stimulation with IFN-γ. Additionally, SHED and OOMDSCs have been shown to have low immunogenicity because they lack expression of HLA-DR and costimulatory molecules such as CD40, CD80, and CD86 before and after IFN-γ treatment. Last, SHED and OOMDSCs expressed the immunoregulatory molecule HLA-G, and the expression of this antigen increased after IFN-γ treatment. In particular, an increase in intracellular HLA-G expression was observed. The results obtained suggest that SHED and OOMDSCs lack immunogenicity and have immunomodulatory properties that are enhanced when they undergo inflammatory stimulation with IFN-γ, which opens new perspectives for the therapeutic use of these cells.
In the present study, hydroxyapatite and Zn-substituted hydroxyapatite powders were synthesized using ostrich eggshell as a calcium source. The samples were analyzed by scanning electron microscopy with field emission gun, and X-ray diffraction (XRD) to identify the present phases, and X-ray fluorescence spectroscopy for quantitative chemical analysis of the synthesized and heat treated powders. The Fourier transform infrared spectroscopy technique was used before and after heat treatments at 700, 900 and 1100 °C in order to identify the functional groups present, as an additional technique to the XRD analysis. The results presented in this study represent a promising method for synthesis of hydroxyapatite and hydroxyapatite partially substituted with zinc, since the results showed no undesirable phases or impurities in the produced powders. It was observed that Zn-substituted hydroxyapatite showed higher thermal stability, when compared to pure hydroxyapatite.
The Hydroxyapatite (HA), the major component of the inorganic portion of bones, may be obtained from different sources for use as biomaterial in the reconstruction of critical bone defects. Among these resources, the eggshell appears as a promising alternative since it is a waste material which may be reused as a source of calcium carbonate after adequate treatment, giving rise to mineral hydroxyapatite. After morphological and physicochemical adjustments allowing its standardization and utilization, hydroxyapatite (HA) may turn into a potentially useful biomaterial for different experimental purposes. This paper aims to present a viable method for obtaining a hydroxyapatite from Ostrich eggshell by wet precipitation process. This phosphating technique with emphasis on the issue of sustainable reuse of disposable materials allows the production of a synthetic crystalline material with characteristics similar to those of bone and tooth. The structural characterization was performed by X-Ray diffraction and scanning electron microscopy with field emission gun and energy dispersive X-ray in the Military Institute of Engineering.
Introduction: Stem cells obtained from the pulp of human deciduous teeth are highly proliferative and plastic multipotent cells, which makes them a relevant model of stem cells, applied in several biomedical areas, with different purposes. Objective: Based on a brief review of the literature, the present work intends to present from conceptual aspects about stem cells, classifications, potential (in vitro and in vivo) applications in dental practice, cell culture, cryopreservation and its importance, ethical and regulatory aspects, as well as the role of the dental surgeon as the endorser responsible for the entire clinical stage that involves the process of collecting stem cells obtained from dental pulps for cryopreservation, with a view to using them under appropriate conditions, in accordance with scientifically proven and justified good laboratory and clinical practices.
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