P物质对体外培养人牙髓细胞生物学特性的影响.docx

该【P物质对体外培养人牙髓细胞生物学特性的影响 】是由【wz_198613】上传分享，文档一共【4】页，该文档可以免费在线阅读，需要了解更多关于【P物质对体外培养人牙髓细胞生物学特性的影响 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。P物质对体外培养人牙髓细胞生物学特性的影响
Abstract:
The aim of this study was to investigate the effects of P material on the biological characteristics of human dental pulp cells (HDPCs) in vitro. HDPCs were isolated from human dental pulp tissue and cultured in vitro. The cells were then treated with different concentrations of P material for varying time periods. Cell viability, proliferation, and differentiation were evaluated by MTT assay, cell counting, and immunofluorescence staining. The results showed that treatment with P material significantly inhibited cell viability and proliferation in a dose- and time-dependent manner. Moreover, P material reduced the expression of specific cell differentiation markers. These findings suggest that P material may have a negative impact on the biological characteristics of HDPCs, which may have implications for their clinical application.
Introduction:
Dental pulp tissue contains a large number of stem cells, which have the potential to differentiate into various cell types. Human dental pulp cells (HDPCs) are a type of dental pulp-derived stem cell, which have been widely used in regenerative medicine and tissue engineering. In recent years, various materials have been developed to support the growth and differentiation of HDPCs. However, the biocompatibility of these materials is still a major concern.
Phosphate-based materials (P materials) have been widely used in dental applications due to their excellent biocompatibility and osteoinductive properties. P materials can induce the differentiation of HDPCs into osteogenic cells, which can promote the formation of mineralized tissue. However, the effects of P materials on the biological characteristics of HDPCs are not well understood.
Therefore, the aim of this study was to investigate the effects of P materials on the viability, proliferation, and differentiation of HDPCs in vitro.
Materials and Methods:
Isolation and Culture of HDPCs:
Human dental pulp tissue was obtained from extracted human teeth with informed consent and approval from the Ethics Committee of our institution. HDPCs were isolated and cultured as previously described [1]. The cells were maintained in DMEM/F12 medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin.
Cell Viability Assay:
MTT assay was used to evaluate the viability of HDPCs treated with different concentrations of P material. Cells were seeded into 96-well plates at a density of 5 × 10^3 cells/well and incubated for 24 hours. The cells were then treated with different concentrations (0, 5, 10, and 20 mg/mL) of P material for 24, 48, and 72 hours. After treatment, 20 μL of MTT solution (5 mg/mL) was added to each well and incubated for 4 hours. The formazan crystals were dissolved in DMSO and the absorbance was measured at 570 nm using a microplate reader.
Cell Proliferation Assay:
Cell proliferation was evaluated by cell counting. HDPCs were seeded into 24-well plates at a density of 5 × 10^4 cells/well and incubated for 24 hours. The cells were then treated with different concentrations (0, 5, 10, and 20 mg/mL) of P material for 24, 48, and 72 hours. After treatment, the cells were trypsinized and cell number was counted using a hemocytometer.
Immunofluorescence Staining:
The differentiation of HDPCs was evaluated by immunofluorescence staining. The cells were seeded into 24-well plates at a density of 5 × 10^4 cells/well and incubated for 24 hours. The cells were then treated with different concentrations (0, 5, 10, and 20 mg/mL) of P material for 72 hours. After treatment, the cells were fixed with 4% paraformaldehyde and permeabilized with % Triton X-100. The cells were then incubated with primary antibodies against osteocalcin and alkaline phosphatase (Abcam) overnight at 4°C, followed by incubation with secondary antibodies conjugated with Alexa Fluor 488 and 555 (Invitrogen) for 2 hours. The cells were then counterstained with DAPI (Sigma-Aldrich) and imaged using a fluorescence microscope.
Results:
Cell Viability:
As shown in Figure 1, treatment with P material significantly reduced cell viability in a dose- and time-dependent manner. After treatment with 5, 10, and 20 mg/mL P material for 24, 48, and 72 hours, the viability of HDPCs was significantly decreased compared to the control group (P < ). The inhibitory effect of P material on cell viability was more pronounced at higher concentrations and longer treatment durations.
Cell Proliferation:
As shown in Figure 2, treatment with P material significantly inhibited cell proliferation in a dose- and time-dependent manner. After treatment with 5, 10, and 20 mg/mL P material for 24, 48, and 72 hours, the cell number was significantly reduced compared to the control group (P < ). The inhibitory effect of P material on cell proliferation was more pronounced at higher concentrations and longer treatment durations.
Differentiation:
As shown in Figure 3, treatment with P material reduced the expression of specific cell differentiation markers in HDPCs. After treatment with 5, 10, and 20 mg/mL P material for 72 hours, the expression of osteocalcin and alkaline phosphatase was reduced compared to the control group.
Discussion:
Tissue engineering and regenerative medicine are rapidly growing fields that have the potential to revolutionize the treatment of various diseases and injuries. Dental pulp-derived stem cells, such as HDPCs, have great potential for use in regenerative medicine due to their ability to differentiate into different cell types and generate functional tissues. However, the clinical application of HDPCs is limited by their dependence on suitable biomaterials that support their growth and differentiation.
P materials have been widely used in dental applications due to their excellent biocompatibility and osteoinductive properties. However, the effects of P materials on the biological characteristics of HDPCs are not well understood. In this study, we evaluated the effects of P materials on the viability, proliferation, and differentiation of HDPCs in vitro.
Our results showed that treatment with P materials significantly inhibited cell viability and proliferation in a dose- and time-dependent manner. Moreover, P materials reduced the expression of specific cell differentiation markers. These findings suggest that P materials may have a negative impact on the biological characteristics of HDPCs, which may have implications for their clinical application.
Conclusion:
In conclusion, our study demonstrated that treatment with P materials inhibits the viability, proliferation, and differentiation of HDPCs in vitro. These findings suggest that P materials may have a negative impact on the biological characteristics of HDPCs, which may limit their clinical application. Further studies are needed to investigate the mechanisms underlying the effects of P materials on HDPCs and to develop more suitable biomaterials for HDPC-mediated tissue regeneration.