Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (8): 1193-1198.doi: 10.12307/2023.138

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Calcium phosphate cement/poly(lactic-co-glycolic acid) degradation products promote osteoclast differentiation of mouse monocytes

Long Guiyue1, Li Dongdong1, Liao Hongbing2   

  1. 1Guangxi Key Laboratory of Oral and Maxillofacial Restoration and Reconstruction, Guangxi Cranio-Maxillofacial Malformation Clinical Research Center, Guangxi Health Commission Key laboratory of prevention and treatment for oral infectious diseases, 2Department of Prosthetics, School of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Received:2022-03-09 Accepted:2022-05-14 Online:2023-03-18 Published:2022-07-27
  • Contact: Liao Hongbing, MD, Professor, Department of Prosthetics, School of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Long Guiyue, Master candidate, Guangxi Key Laboratory of Oral and Maxillofacial Restoration and Reconstruction, Guangxi Cranio-Maxillofacial Malformation Clinical Research Center, Guangxi Key Laboratory of Oral and Maxillofacial Surgery, Medical Experimental Center of Guangxi Medical University, School of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 81860201, 81560190 (to LHB)

Abstract: BACKGROUND: Preliminary studies found that calcium phosphate cement/poly(lactic-co-glycolic acid) composite can accelerate the process of bone remodeling, and the effect of its degradation products on the differentiation of mononuclear osteoclasts needs further study.
OBJECTIVE: To observe the effect of calcium phosphate cement/poly(lactic-co-glycolic acid) degradation products on the differentiation of mouse RAW264.7 monocytes and osteoclasts. 
METHODS: The experiment was divided into blank control group, glycolic acid group, and calcium phosphate cement/poly(lactic-co-glycolic acid) group. According to the experimental group assignment, the prepared solution and 50 μg/L ligand of nuclear factor κB receptor activator were added to complete medium to prepare media with different conditions. Mouse RAW264.7 cells were cultured for 5 days to induce their differentiation. Cell counting kit-8 assay was used to analyze the effect of different culture media on cell proliferation. RT-PCR and western blot assay were used to detect the changes of gene and protein expression levels of nuclear factor of activated T-cells cytoplasmic 1, matrix metalloproteinase-9, nuclear factor κB receptor activator and tartrate-resistant acid phosphatase activated by osteoclast differentiation-related factors in mouse RAW264.7 monocytes. The osteoclast-like cells were identified by tartrate-resistant acid phosphatase staining. 
RESULTS AND CONCLUSION: Cell counting kit-8 assay results showed that the cells were in a rapid growth period within 72 hours, and then began to decline after 96 hours. The results of RT-PCR and western blot assay showed that the expression levels of nuclear factor of activated T-cells cytoplasmic 1, nuclear factor-κB receptor activator and tartrate-resistant acid phosphatase mRNA and protein expression levels of nuclear factor-1 and nuclear factor-κB receptor activator in activated T cells in calcium phosphate cement/poly(lactic-co-glycolic acid) group were significantly higher than those in glycolic acid group and control group (P < 0.05). There was no significant difference in gene and protein expression levels of matrix metalloproteinase-9 between glycolic acid group and calcium phosphate cement/poly(lactic-co-glycolic acid) group, but it was higher than that of the control group (P < 0.01). Tartrate-resistant acid phosphatase staining results showed that the number of osteoclast-like cells in the calcium phosphate cement/poly(lactic-co-glycolic acid) group was significantly higher than that in the control group and the glycolic acid group (P < 0.05). Overall, these results indicate that the microenvironment formed by the degradation products of calcium phosphate cement/poly(lactic-co-glycolic acid) can promote the osteoclast differentiation of mouse RAW264.7 monocytes. 

Key words: calcium phosphate cement/poly(lactic-co-glycolic acid) copolymer, monocyte, osteoclast differentation, matrix metalloproteinase-9, nuclear factor of activated T-cells cytoplasmic 1, nuclear factor-κB receptor activator

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