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Cellular response of chondrocytes to magnesium alloys for orthopedic applications.

Liao Y, Xu Q, Zhang J, Niu J, Yuan G, Jiang Y, He Y, Wang X - Int. J. Mol. Med. (2015)

Bottom Line: There were no statistically significant differences observed between the JDBM, AZ31, WE43 and pure Mg group (p>0.05).These findings suggested that the JDBM alloy was highly biocompatible with chondrocytes in vitro, yielding results similar to those of AZ31, WE43 and pure Mg.Furthermore, CaHPO4·2H2O coating significantly improved the biocompatibility of this alloy.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, The Fifth People's Hospital of Shanghai, Fudan University, Shanghai 200240, P.R. China.

ABSTRACT
In the present study, the effects of Mg-Nd-Zn-Zr (JDBM), brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), AZ31, WE43, pure magnesium (Mg) and Ti alloy (TC4) on rabbit chondrocytes were investigated in vitro. Adhesion experiments revealed the satisfactory morphology of chondrocytes on the surface of all samples. An indirect cytotoxicity test using MTT assay revealed that C‑JDBM and TC4 exhibited results similar to those of the negative control, better than those obtained with JDBM, AZ31, WE43 and pure Mg (p<0.05). There were no statistically significant differences observed between the JDBM, AZ31, WE43 and pure Mg group (p>0.05). The results of indirect cell cytotoxicity and proliferation assays, as well as those of apoptosis assay, glycosaminoglycan (GAG) quantification, assessment of collagen Ⅱ (Col Ⅱ) levels and RT-qPCR revealed a similar a trend as was observed with MTT assay. These findings suggested that the JDBM alloy was highly biocompatible with chondrocytes in vitro, yielding results similar to those of AZ31, WE43 and pure Mg. Furthermore, CaHPO4·2H2O coating significantly improved the biocompatibility of this alloy.

No MeSH data available.


Related in: MedlinePlus

Morphology of the chondrocytes follwoing 1, 3 and 5 days of incubation with: (A–C and M) Dulbecco’s modified Eagle’s medium with F12 (DMEM/F12), (D and P) 10% DMSO medium, (E–G and N) brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), (H and O) Ti alloy (TC4), (I and Q) Mg-Nd-Zn-Zr (JDBM), (J and R) AZ31, (K and S) WE43, and (L and T) pure magnesium (Mg). (A–C and E–G) Cells were incubated for 1, 3 and 5 days, respectively. (D and H–T) Cells were incubated for 3 days. (A–L) Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (x40 magnification). (M–T) Cells were stained with Alcian blue (x400 magnification). A greater number of cells was observed in the negative control group (B), C-JDBM (F) and TC4 (H) groups after 3 days, as compared with the other groups (I–L), due to the loss of cells in the center of the other culture plates, while no other differences were observed between them. (A–C and E–G) The number of cells was directly associated with the incubation time. The chondrocytes in the positive control group were extremely scarce (D and P), and unlike the elongated, polygonal, deltoid or irregular shape of the other chondrocytes (M–O and Q–T), their morphology had changed to a small and round shape (P), whereas the cells from all the other treatment groups appeared normal.
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f4-ijmm-36-01-0073: Morphology of the chondrocytes follwoing 1, 3 and 5 days of incubation with: (A–C and M) Dulbecco’s modified Eagle’s medium with F12 (DMEM/F12), (D and P) 10% DMSO medium, (E–G and N) brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), (H and O) Ti alloy (TC4), (I and Q) Mg-Nd-Zn-Zr (JDBM), (J and R) AZ31, (K and S) WE43, and (L and T) pure magnesium (Mg). (A–C and E–G) Cells were incubated for 1, 3 and 5 days, respectively. (D and H–T) Cells were incubated for 3 days. (A–L) Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (x40 magnification). (M–T) Cells were stained with Alcian blue (x400 magnification). A greater number of cells was observed in the negative control group (B), C-JDBM (F) and TC4 (H) groups after 3 days, as compared with the other groups (I–L), due to the loss of cells in the center of the other culture plates, while no other differences were observed between them. (A–C and E–G) The number of cells was directly associated with the incubation time. The chondrocytes in the positive control group were extremely scarce (D and P), and unlike the elongated, polygonal, deltoid or irregular shape of the other chondrocytes (M–O and Q–T), their morphology had changed to a small and round shape (P), whereas the cells from all the other treatment groups appeared normal.

Mentions: The absorbance of the chondrocytes cultured on the different extracts, DMEM/F12 and 10% DMSO media for 1, 3, and 5 days is shown in Fig. 3. The morphologies of the chondrocytes after 1, 3, and 5 days of incubation are presented in Fig. 4. As shown in Fig. 3, the absorbance of all groups of cells increased with time. As shown in Table I, the time effect (day) and the effects of day x group interaction were statistically significant (p<0.05), indicating that the research target changed with time. Furthermore, the role of the time factor varied within each group. Tests on the between-subject effects indicated that the grouping factor influenced the results, and each group of targets generally differed (p<0.05). Likewise, the absorbance of C-JDBM (Fig. 3) was greater than that of the negative control and TC4, although the difference was not significant (p>0.05). The absorbance of the remaining groups showed significantly lower results (p<0.05). By contrast, no significant differences were observed among the JDBM, AZ31, WE43 and pure Mg groups (p>0.05), thereby indicating that these alloys have high cytocompatibility with chondrocytes.


Cellular response of chondrocytes to magnesium alloys for orthopedic applications.

Liao Y, Xu Q, Zhang J, Niu J, Yuan G, Jiang Y, He Y, Wang X - Int. J. Mol. Med. (2015)

Morphology of the chondrocytes follwoing 1, 3 and 5 days of incubation with: (A–C and M) Dulbecco’s modified Eagle’s medium with F12 (DMEM/F12), (D and P) 10% DMSO medium, (E–G and N) brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), (H and O) Ti alloy (TC4), (I and Q) Mg-Nd-Zn-Zr (JDBM), (J and R) AZ31, (K and S) WE43, and (L and T) pure magnesium (Mg). (A–C and E–G) Cells were incubated for 1, 3 and 5 days, respectively. (D and H–T) Cells were incubated for 3 days. (A–L) Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (x40 magnification). (M–T) Cells were stained with Alcian blue (x400 magnification). A greater number of cells was observed in the negative control group (B), C-JDBM (F) and TC4 (H) groups after 3 days, as compared with the other groups (I–L), due to the loss of cells in the center of the other culture plates, while no other differences were observed between them. (A–C and E–G) The number of cells was directly associated with the incubation time. The chondrocytes in the positive control group were extremely scarce (D and P), and unlike the elongated, polygonal, deltoid or irregular shape of the other chondrocytes (M–O and Q–T), their morphology had changed to a small and round shape (P), whereas the cells from all the other treatment groups appeared normal.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494570&req=5

f4-ijmm-36-01-0073: Morphology of the chondrocytes follwoing 1, 3 and 5 days of incubation with: (A–C and M) Dulbecco’s modified Eagle’s medium with F12 (DMEM/F12), (D and P) 10% DMSO medium, (E–G and N) brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), (H and O) Ti alloy (TC4), (I and Q) Mg-Nd-Zn-Zr (JDBM), (J and R) AZ31, (K and S) WE43, and (L and T) pure magnesium (Mg). (A–C and E–G) Cells were incubated for 1, 3 and 5 days, respectively. (D and H–T) Cells were incubated for 3 days. (A–L) Cells were stained with 4′,6-diamidino-2-phenylindole (DAPI) (x40 magnification). (M–T) Cells were stained with Alcian blue (x400 magnification). A greater number of cells was observed in the negative control group (B), C-JDBM (F) and TC4 (H) groups after 3 days, as compared with the other groups (I–L), due to the loss of cells in the center of the other culture plates, while no other differences were observed between them. (A–C and E–G) The number of cells was directly associated with the incubation time. The chondrocytes in the positive control group were extremely scarce (D and P), and unlike the elongated, polygonal, deltoid or irregular shape of the other chondrocytes (M–O and Q–T), their morphology had changed to a small and round shape (P), whereas the cells from all the other treatment groups appeared normal.
Mentions: The absorbance of the chondrocytes cultured on the different extracts, DMEM/F12 and 10% DMSO media for 1, 3, and 5 days is shown in Fig. 3. The morphologies of the chondrocytes after 1, 3, and 5 days of incubation are presented in Fig. 4. As shown in Fig. 3, the absorbance of all groups of cells increased with time. As shown in Table I, the time effect (day) and the effects of day x group interaction were statistically significant (p<0.05), indicating that the research target changed with time. Furthermore, the role of the time factor varied within each group. Tests on the between-subject effects indicated that the grouping factor influenced the results, and each group of targets generally differed (p<0.05). Likewise, the absorbance of C-JDBM (Fig. 3) was greater than that of the negative control and TC4, although the difference was not significant (p>0.05). The absorbance of the remaining groups showed significantly lower results (p<0.05). By contrast, no significant differences were observed among the JDBM, AZ31, WE43 and pure Mg groups (p>0.05), thereby indicating that these alloys have high cytocompatibility with chondrocytes.

Bottom Line: There were no statistically significant differences observed between the JDBM, AZ31, WE43 and pure Mg group (p>0.05).These findings suggested that the JDBM alloy was highly biocompatible with chondrocytes in vitro, yielding results similar to those of AZ31, WE43 and pure Mg.Furthermore, CaHPO4·2H2O coating significantly improved the biocompatibility of this alloy.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopaedics, The Fifth People's Hospital of Shanghai, Fudan University, Shanghai 200240, P.R. China.

ABSTRACT
In the present study, the effects of Mg-Nd-Zn-Zr (JDBM), brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), AZ31, WE43, pure magnesium (Mg) and Ti alloy (TC4) on rabbit chondrocytes were investigated in vitro. Adhesion experiments revealed the satisfactory morphology of chondrocytes on the surface of all samples. An indirect cytotoxicity test using MTT assay revealed that C‑JDBM and TC4 exhibited results similar to those of the negative control, better than those obtained with JDBM, AZ31, WE43 and pure Mg (p<0.05). There were no statistically significant differences observed between the JDBM, AZ31, WE43 and pure Mg group (p>0.05). The results of indirect cell cytotoxicity and proliferation assays, as well as those of apoptosis assay, glycosaminoglycan (GAG) quantification, assessment of collagen Ⅱ (Col Ⅱ) levels and RT-qPCR revealed a similar a trend as was observed with MTT assay. These findings suggested that the JDBM alloy was highly biocompatible with chondrocytes in vitro, yielding results similar to those of AZ31, WE43 and pure Mg. Furthermore, CaHPO4·2H2O coating significantly improved the biocompatibility of this alloy.

No MeSH data available.


Related in: MedlinePlus