Evaluation of biological properties of Gd-doped hydroxyapatite bio-nanocomposites

KONG Wei-li YANG Yu SHEN Fu-guo SUN Wen-cai GU Hao JIN Song XIAO Wen-long

Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (5) : 632-640.

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Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (5) : 632-640. DOI: 10.16098/j.issn.0529-1356.2024.05.016
Bioengineering

Evaluation of biological properties of Gd-doped hydroxyapatite bio-nanocomposites

  • KONG  Wei-li1  YANG  Yu3  SHEN  Fu-guo2*  SUN  Wen-cai2  GU  Hao2  JIN  Song2  XIAO  Wen-long
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Abstract

Objective  To investigate the biocompatibility of new gadolinium-doped hydroxyapatite (Gd-HA) composite scaffolds and to explore their feasibility as cell culture materials and bone tissue engineering scaffolds.Methods  The Gd-HA composite scaffolds were chemically synthesized and placed under the electron microscope for observation. The experiment was divided into three groups, the HA group, the Gd-HA group, and the control group.Rabbit adipose-derived mesenchymal stem cells (ADSCs) were isolated, cultured and characterized, and the Gd-HA composite scaffold extract was added to the ADSCs in vitro culture system. Cell survival and cytotoxicity were assessed by live-dead cell staining, cell proliferation ability within the scaffolds was assessed by CCK-8 assay, and the scaffolds were assessed by alizarin red staining for cell osteogenic differentiation. The toxic reactions of the scaffold materials were observed by skin irritation test, systemic acute toxicity test and muscle tissue and liver and kidney pathology at the site of intramuscular implantation of the scaffolds.  Results  The Gd-HA composite scaffold showed irregular void structure under electron microscope. Cell morphology observation showed that ADSCs grew adherently to the wall and were long shuttle-shaped. The positivity rate of CD29 was 96.94%, CD44 was 97.90%, CD45 was 0.10%, and CD34 was 0.46%, which was obtained using flow cytometry. Live-dead cell staining showed that the amount of live cells in the Gd-HA group was significantly better than that in the hydroxyapatite(HA) group after 5 days of co-culture. CCK-8 assay showed no significant difference in cell proliferation within 0-3days. After 3days, the Gd-HA group was significantly better than the HA group and the control group (P<0.05). Calcium nodule deposition after alizarin red staining was significantly better in the Gd-HA group than in the HA and control groups, showing a deeper red color. No skin irritation was observed in gross and skin tissue HE observations after the contact of the extract with the skin. The general condition of the experimental groups was good after the infusion of the extract into the abdominal cavity, and the body mass tended to increase steadily (P>0.05). HE staining showed that inflammatory reaction at the interface between the material and muscle tissue of the stent intramuscular implantation site in Gd-HA group was significantly higher than that of the control group, and the inflammatory cell infiltration was gradually reduced with the prolongation of implantation time. At the 8th weeks the morphology of the tissue around the material was close to normal muscle tissue, and no pathological changes were observed in the HE staining of liver and kidney at the 12th week. Conclusion   Gd-HA composite scaffolds exhibit good biocompatibility and facilitate cell proliferation and osteogenic differentiation, and they are expected to serve as good carriers for stem cell transplantation in tissue engineering. 

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KONG Wei-li YANG Yu SHEN Fu-guo SUN Wen-cai GU Hao JIN Song XIAO Wen-long. Evaluation of biological properties of Gd-doped hydroxyapatite bio-nanocomposites[J]. Acta Anatomica Sinica. 2024, 55(5): 632-640 https://doi.org/10.16098/j.issn.0529-1356.2024.05.016

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