Regulation of Tendon-bone Related Multiple Cells by Chemical Ions Released from Mo-containing Silicate Bioceramics★

doi:10.6023/A23040120

Abstract

Understanding the relationship between the chemical signals generated by biological materials and cellular behaviors has great significance for the design and preparation of high-performance tissue engineering biomaterials. In the past several decades, silicate bioceramics have been widely used in tissue engineering. Bioactive ions released from silicate bioceramics can act as chemical signals to regulate cellular behaviors and promote the tissue regeneration. Moreover, by regulating the components of silicate bioceramics, silicate bioceramics can generate specific chemical signals to regulate cellular behaviors of multiple cells. Here, by introducing molybdenum (Mo) element into silicate bioceramics, we have successfully developed Mo-containing silicate (MS) bioceramics which are able to regulate cellular behaviors of tendon stem/progenitor cells (TSPCs) and bone marrow mesenchymal stem cells (BMSCs) simultaneously. Using ammonium molybdate as a source of Mo element, MS bioceramics were prepared by chemical coprecipitation method. The synthesized MS bioceramics were mostly below 10 μm in size and had uniform distribution of Mo elements. Moreover, MS bioceramics were composed of high-purity CaMoO₄ and CaSiO₃. To explore the effect of chemical signals generated from MS bioceramics on TSPCs and BMSCs, we prepared MS extracts for cell culture. MS bioceramics supported the survival of TSPCs and BMSCs and maintained a better cellular state during the culture period of 5 d. Due to the released Ca, Si and Mo ions from MS bioceramics, TSPCs and BMSCs cultured with MS extracts exhibited excellent proliferation and migration activities. Interestingly, after cultured with MS extracts in the appropriate concentration, the expression of osteogenic genes and protein of BMSCs and the expression of tenogenic genes and protein of TSPCs were significantly enhanced, suggesting that chemical signals generated by MS bioceramics simultaneously promoted the specific differentiation of TSPCs and BMSCs. Such MS bioceramics are believed to be an effective “bioactive factor” for repairing injury at the tendon-bone interfaces.

Key words: Mo-containing silicate, chemical signals, multiple cells, cellular behaviors

Cite this article

Lin Du, Jianmin Xue, Zhiguang Huan, Chengtie Wu. Regulation of Tendon-bone Related Multiple Cells by Chemical Ions Released from Mo-containing Silicate Bioceramics^★[J]. Acta Chimica Sinica, 2023, 81(10): 1334-1340.

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Fig. & Tab. 7

Figure 1. Characterization of Mo-containing silicate (MS) bioceramic powders. (A) XRD pattern of MS particles. (B) SEM image of MS particles. (C) Particle size distribution of MS particles. (D) EDS elemental mapping of MS particles. MS bioceramics were successfully prepared with uniform particle size and homogeneous distribution of Mo element

Ionic conc./(mg•L^－1)	Ca	Si	Mo
Control	71.85±2.58	2.45±1.13	0±0
1/2	83.77±4.34	136.06±7.61	14.29±0.94
1/4	96.18±6.39	73.19±4.80	7.67±0.62
1/8	85.99±2.43	39.29±0.85	3.92±0.31
1/16	79.29±4.85	21.09±0.89	1.95±0.07
1/32	76.39±2.83	11.75±0.20	1.27±0.08
1/64	70.53±1.67	6.96±0.35	0.43±0.26
1/128	72.99±0.36	4.86±0.21	0.37±0.13
1/256	70.92±1.82	3.40±0.94	0.18±0.22

Table 1. The concentration of chemical signals generated from different concentrations of MS extracts

Figure 2. Proliferation and live/dead assay of TSPCs (tendon stem/progenitor cells) and BMSCs (bone marrow mesenchymal stem cells) cultured with different concentrations of MS extracts. Proliferative activity of (A) TSPCs and (B) BMSCs after cultured with different dilution of MS extracts (n=3) *p<0.05, **p<0.01, ***p<0.001. Live/dead assay of TSPCs (C) and BMSCs (D) after cultured with MS extracts for 1 and 5 d

Figure 3. The effect of different dilution of MS extracts on migration of TSPCs and BMSCs. After induced with MS extracts, crystalline violet staining images of the migration of TSPCs (A) and BMSCs (B). Quantification of TSPCs (C) and BMSCs (D) induced to migrate (n=4). *p<0.05, **p<0.01, ***p<0.001

Figure 4. The expression of tenogenic differentiation-related genes and Tenomodulin (TNMD) of TSPCs cultured with different dilutions of MS extracts (1/16 and 1/32). The expression of (A) BGN, (B) DCN and (C) TNC of TSPCs incubated with different dilutions of MS extracts after 5 days of culture (n=6). *p<0.05, **p<0.01, ***p<0.001. (D) Immunofluorescence staining images of TNMD protein of TSPCs cultured with different concentrations MS extracts

Figure 5. The expression of osteogenic differentiation-related genes and Bone sialoprotein (BSP) of BMSCs cultured with different dilutions of MS extracts. (A) The expression of ALP, (B) BMP2, (C) Run×2 and (D) OPN of BMSCs incubated with different dilution of MS extracts after 5 days of culture (n=6). *p<0.05, **p<0.01, ***p<0.001. (E) Immunofluorescence staining images of BSP protein of BMSCs cultured with different concentrations of MS extracts

Figure 6. Semi-quantitative analysis of immunofluorescence staining images of TNMD (A) and BSP (B) expression (n=3). *p<0.05, **p<0.01, ***p<0.001

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含钼的硅酸盐生物陶瓷释放的化学离子对肌腱-骨相关的多细胞调控^★