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Open Access Basic Study Issue
Effect of angiopoietin 4 on odontogenic differentiation of dental pulp stem cells
Journal of Prevention and Treatment for Stomatological Diseases 2023, 31(10): 692-700
Published: 20 October 2023
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Objective

To investigate the effects of angiopoietin 4 (ANGPT4) on the odontogenic differentiation of human dental pulp stem cells.

Methods

This study has been reviewed and approved by the Ethics Committee, and informed consent has been obtained from patients. Human premolars were fixed, decalcified, dehydrated, embedded, and sectioned. Immunofluorescence staining was used to observe the expression and localization of ANGPT4. Human dental pulp stem cells (hDPSCs) were isolated and cultured in vitro. The growth state and morphology of hDPSCs were observed under an inverted phase contrast microscope. The expression of cell surface-related molecular markers was detected by flow cytometry. Alkaline phosphatase and alizarin red S staining were used to detect the odontogenic differentiation potential of hDPSCs. Oil-red O staining was used to detect the adipogenic differentiation potential of hDPSCs. RNA was extracted from hDPSCs at different time points after odontogenic induction, and RT-qPCR was used to analyze the mRNA expression of ANGPT4 and odontogenic-related genes during the odontogenic differentiation of hDPSCs in vitro. siRNA gene silencing technology was used to silence the expression of ANGPT4 in hDPSCs, and the silencing efficiency was detected by RT-qPCR and Western Blot. After silencing ANGPT4 in hDPSCs for 24 h, odontogenic induction was performed. Alkaline phosphatase and alizarin red S staining were performed on the 7th and 14th of induction to detect the odontogenic differentiation ability of hDPSCs after silencing ANGPT4.

Results

Immunofluorescence staining of human premolars showed that ANGPT4 was expressed in odontoblasts and sub-odontoblastic cell-rich zone. hDPSCs were in good condition after 14 days of isolation and culture. Under the microscope, multiple cell colonies were observed, and the cell morphology was uniform and long spindle-shaped. The results of flow cytometry showed that hDPSCs expressed mesenchymal stem cell markers CD105 (90.42%) and CD90 (97.15%), but did not express hematopoietic cell markers CD45 (0.01%) and CD34 (0.08%). After odontogenic and adipogenic induction of hDPSCs, alkaline phosphatase staining, alizarin red S staining and oil red O staining were positive. The results of RT-qPCR after the odontogenic induction of hDPSCs showed that ANGPT4 was highly expressed on the 5th, 7th, 11th and 14th days of differentiation of hDPSCs (P < 0.05), with the highest expression level on the 5th day. After hDPSCs were transfected with si-ANGPT4, the expression of ANGPT4 mRNA and protein was significantly down-regulated (P < 0.05). The results of alkaline phosphatase staining showed that ALP staining intensity and area in the si-ANGPT4 group were significantly lower than those in the negative control. Alizarin red S staining showed that the formation of calcium nodules in the si-ANGPT4 group was significantly lower than that in the negative control.

Conclusion

ANGPT4 was expressed in odontoblasts and sub-odontoblastic cell-rich zone of human premolars. ANGPT4 may be a factor to promote the odontogenic differentiation of hDPSCs.

Open Access Review Article Issue
Research progress on the mechanical compatibility of restorative materials with dental tissue
Journal of Prevention and Treatment for Stomatological Diseases 2024, 32(4): 315-320
Published: 20 April 2024
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As the main means of mastication, teeth can withstand countless functional contacts. The mechanical properties of teeth are closely related to their tissue structure. Enamel and dentin have a high hardness and modulus of elasticity, and their graded structure allows them to withstand bite forces without being susceptible to fracture. When tooth tissue is defective, full crown restoration is often needed to restore the normal shape and function of the tooth. Metal materials, ceramic materials, and polyetheretherketone (PEEK) materials are commonly used for crown restoration. Metal materials have certain disadvantages in terms of aesthetics and are relatively rarely used in clinical practice. Ceramic materials with different compositions exhibit differences in performance and aesthetics, but their elastic modulus and hardness are much higher than those of dental tissue, resulting in mismatching mechanical properties. In contrast, the elastic modulus of PEEK is lower than that of tooth tissue and similar to that of bone tissue, but its properties can be improved by fiber reinforcement. Notably, when the mechanical properties of a restoration material and tooth tissue are not fully matched, the interface between them often forms a potential weak link, which ultimately affects the stability and long-term effect of the restoration. This article introduces the mechanical properties and corresponding structural characteristics of enamel and dentin. On this basis, the advantages and limitations of existing restoration materials are analyzed, and the possibility of biomimetic design of full crowns is further explored.

Open Access Review Article Issue
Advances in oral mesenchymal stem cell-derived extracellular vesicles in health and disease
Genes & Diseases 2024, 11(1): 346-357
Published: 12 April 2023
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Extracellular vesicles (EVs) are nano-size vesicles secreted naturally by all cells into the extracellular space and have been recognized as important cell–cell mediators in multicellular organisms. EVs contain nucleic acids, proteins, lipids, and other cellular components, regulating many basic biological processes and playing an important role in regenerative medicine and diseases. EVs can be traced to their cells of origin and exhibit a similar function. Moreover, EVs demonstrate low immunogenicity, good biocompatibility, and fewer side effects, compared to their parent cells. Mesenchymal stem cells (MSCs) are one of the most important resource cells for EVs, with a great capacity for self-renewal and multipotent differentiation, and play an essential role in stem cell therapy. The mechanism of MSC therapy was thought to be attributed to the differentiation of MSCs after targeted migration, as previously noted. However, emerging evidence shows the previously unknown role of MSC-derived paracrine factors in stem cell therapy. Especially EVs derived from oral tissue MSCs (OMSC-EVs), show more advantages than those of all other MSCs in tissue repair and regeneration, due to their lower invasiveness and easier accessibility for sample collection. Here, we systematically review the biogenesis and biological characteristics of OMSC-EVs, as well as the role of OMSC-EVs in intercellular communication. Furthermore, we discuss the potential therapeutic roles of OMSC-EVs in oral and systemic diseases. We highlight the current challenges and future directions of OMSC-EVs to focus more attention on clinical translation. We aim to provide valuable insights for the explorative clinical application of OMSC-EVs.

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