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Journal of Peking University (Health Sciences) ›› 2023, Vol. 55 ›› Issue (1): 44-51. doi: 10.19723/j.issn.1671-167X.2023.01.007

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Effect of pH on the chelation between strontium ions and decellularized small intestinal submucosal sponge scaffolds

Yu-ke LI1,Mei WANG2,Lin TANG1,Yu-hua LIU1,*(),Xiao-ying CHEN1   

  1. 1. Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
    2. Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2022-10-12 Online:2023-02-18 Published:2023-01-31
  • Contact: Yu-hua LIU E-mail:liuyuhua@bjmu.edu.cn

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Abstract:

Objective: To investigate the preparation of decellularized small intestinal submucosa (dSIS) sponge scaffolds with chelated strontium (Sr) ions at different pH values, and to select the appropriate pH values for synthesizing Sr/dSIS scaffolds using the physicochemical properties and biocompatibility of the scaffolds as evaluation indexes. Methods: (1) Sr/dSIS scaffolds preparation and grouping: After mixing dSIS solution and strontium chloride solution in equal volumes, adjusting pH of the solution to 3, 5, 7, and 9 respectively, porous scaffolds were prepared by freeze-drying method after full reaction at 37℃, which were named Sr/dSIS-3, -5, -7, and -9 respectively, and the dSIS scaffolds were used as the control group. (2) Physicochemical property evaluation: The bulk morphology of the scaffolds was observed in each group, the microscopic morphology analyzed by scanning electron microscopy, and the porosity and pore size determined, the surface elements analyzed by energy spectroscopy, the structure of functional groups analyzed by infrared spectroscopy, the chelation rate determined by atomic spectrophotometry, the water absorption rate detected by using specific gravity method, and the compression strength evaluated by universal mechanical testing machine.(3) Biocompatibility evaluation: The cytotoxicity and proliferative effect to bone mesenchymal stem cells (BMSCs) of each group were evaluated by Calcein-AM/PI double staining method. Results: Scanning electron microscopy showed that the scaffolds of each group had an interconnected three-dimensional porous structure with no statistical difference in pore size and porosity. Energy spectrum analysis showed that strontium could be detected in Sr/dSIS-5, -7 and -9 groups, and strontium was uniformly distributed in the scaffolds. Functional group analysis further supported the formation of chelates in the Sr/dSIS-5, -7 and -9 groups. Chelation rate analysis showed that the Sr/dSIS-7 group had the highest strontium chelation rate, which was statistically different from the other groups (P < 0.05). The scaffolds in all the groups had good water absorption. The scaffolds in Sr/dSIS-5, -7 and -9 groups showed significantly improved mechanical properties compared with the control group (P < 0.05). The scaffolds in all the groups had good biocompatibility, and the Sr/dSIS-7 group showed the best proliferation of BMSCs. Conclusion: When pH was 7, the Sr/dSIS scaffolds showed the highest strontium chelation rate and the best proliferation effect of BMSCs, which was the ideal pH value for the preparation of the Sr/dSIS scaffolds.

Key words: Decellularized small intestinal submucosa, Strontium, Bone, Tissue engineering, Scaffolds

CLC Number: 

  • R318.08

Figure 1

Macroscopic morphology of the dSIS and different Sr/dSIS scaffolds Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 2

Surface microscopic morphology of the dSIS and Sr/dSIS scaffolds A-E, scanning electron microscopy images of surface morphology (magnification=200); F-J, magnificated images (magnification=500). Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 3

Pore size (A) and porosity (B) of the dSIS and Sr/dSIS scaffolds Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 4

Surface EDX spectra and Mapping of the dSIS and Sr/dSIS scaffolds A, surface EDX spectra of the dSIS and Sr/dSIS scaffolds; B, Mapping of the dSIS and Sr/dSIS scaffolds. EDX, energy dispersive X-ray spectroscopy; Sr, strontium; dSIS, decellularized small intestinal submucosa; C, carbon; N, nitrogen; O, oxygen; CPS, counts per second."

Figure 5

FTIR spectra of the dSIS and Sr/dSIS scaffolds FTIR, flourier transformed infrared spectroscopy; Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 6

Strontium chelation rate of the Sr/dSIS scaffolds * P < 0.05; Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 7

Water absorption of the dSIS and Sr/dSIS scaffolds Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 8

Compressive modulus of the dSIS and Sr/dSIS scaffolds * P < 0.05. Sr, strontium; dSIS, decellularized small intestinal submucosa."

Figure 9

Images of Calcein-AM/PI double staining after 1, 3 and 5 days of BMSCs culture in the dSIS and Sr/dSIS scaffolds Sr, strontium; dSIS, decellularized small intestinal submucosa; BMSCs, bone mesenchymal stem cells."

Figure 10

Percentage of live/dead cells after 1, 3 and 5 days of BMSCs culture in the dSIS and Sr/dSIS scaffolds Sr, strontium; dSIS, decellularized small intestinal submucosa; BMSCs, bone mesenchymal stem cells."

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