Biocompatibility and Cu ions release kinetics of copper-bearing titanium alloys

doi:10.1016/j.jmst.2021.03.074

Abstract

Abstract:

To reduce the risk of implant-associated infections, we previously designed and developed a series of medical copper (Cu)-bearing titanium alloys that release Cu ions and hence play an antibacterial role. However, both excessive and deficient Cu levels adversely affect human health; therefore, the aim of the present study was to comprehensively evaluate the short- and long-term biosafety of Cu-bearing titanium alloys (Ti6Al4V-Cu and Ti-Cu) both in vitro and in vivo. Moreover, the predominant kinetic mechanism of Cu ions release and its effect on biosafety were also investigated. The results indicate that the biocompatibility of the Cu-bearing titanium alloys meets the requirements of ISO standards and the Cu ion release kinetics display a good correlation over the entire time period in the normal zero-order model with an almost constant release rate. The release rate maintained at a parts per billion level safe for humans; consequently, we can conclude that our Cu-bearing titanium alloys have satisfactory biocompatibility.

Key words: Cu ions, Kinetic, Biocompatibility, Titanium

Ling Ren, Xiaohe Xu, Hui Liu, Ke Yang, Xun Qi. Biocompatibility and Cu ions release kinetics of copper-bearing titanium alloys[J]. J. Mater. Sci. Technol., 2021, 95: 237-248.

Figures/Tables 14

References 34

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Experiments	Samples	Size	Extract medium	Conditions
MTT assay	Ti6Al4V-Cu	Φ11.25 mm × 1.37mm	89% MEM, 10% FBS, and 1% p/s	37 °C, 60 rpm, 24 h
	TiCu	Φ10.00 mm × 1.05mm
In vitro mammalian TK gene mutation assay	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: RPMI 1640 Non-polar: DMSO	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.00mm
In vitro mammalian chromosome aberration assay	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: RPMI 1640 Non-polar: DMSO	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.00mm
Bacterial reverse mutation assay (Ames test)	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: Normal Saline Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ10.00 mm × 1.00mm
Acute systemic toxicity assay	Ti6Al4V-Cu	Φ11.25 mm × 1.37mm	Polar: Normal Saline
Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ10.00 mm × 1.05mm
Subchronic systemic toxicity assay	Ti6Al4V-Cu	Φ10.50 mm × 2.20mm	Polar: Normal Saline Non-polar: sesame oil	37 °C, 60 rpm, 72 h
	TiCu	Φ11.50 mm × 2.20mm
Skin irritation study	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: Normal Saline Non-polar: cottonseed oil	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.05mm
Skin sensitization study	Ti6Al4V-Cu	Φ10.18 mm × 1.37mm	Polar: Normal Saline Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ11.14 mm × 2.08mm
Pyrogen assay	Ti6Al4V-Cu	Φ10.50 mm × 1.90mm	Normal Saline	37 °C, 60 rpm, 72 h
	TiCu	Φ11.46 mm × 2.13mm

Experiments	Samples	Size	Extract medium	Conditions
MTT assay	Ti6Al4V-Cu	Φ11.25 mm × 1.37mm	89% MEM, 10% FBS, and 1% p/s	37 °C, 60 rpm, 24 h
	TiCu	Φ10.00 mm × 1.05mm
In vitro mammalian TK gene mutation assay	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: RPMI 1640 Non-polar: DMSO	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.00mm
In vitro mammalian chromosome aberration assay	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: RPMI 1640 Non-polar: DMSO	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.00mm
Bacterial reverse mutation assay (Ames test)	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: Normal Saline Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ10.00 mm × 1.00mm
Acute systemic toxicity assay	Ti6Al4V-Cu	Φ11.25 mm × 1.37mm	Polar: Normal Saline
Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ10.00 mm × 1.05mm
Subchronic systemic toxicity assay	Ti6Al4V-Cu	Φ10.50 mm × 2.20mm	Polar: Normal Saline Non-polar: sesame oil	37 °C, 60 rpm, 72 h
	TiCu	Φ11.50 mm × 2.20mm
Skin irritation study	Ti6Al4V-Cu	Φ10.00 mm × 1.00mm	Polar: Normal Saline Non-polar: cottonseed oil	37 °C, 60 rpm, 72 h
	TiCu	Φ10.00 mm × 1.05mm
Skin sensitization study	Ti6Al4V-Cu	Φ10.18 mm × 1.37mm	Polar: Normal Saline Non-polar: cottonseed oil	121 °C, 1 h
	TiCu	Φ11.14 mm × 2.08mm
Pyrogen assay	Ti6Al4V-Cu	Φ10.50 mm × 1.90mm	Normal Saline	37 °C, 60 rpm, 72 h
	TiCu	Φ11.46 mm × 2.13mm

Sample	Extract Liquid	S9	Time point	Index	Negative control	25%	50%	100%	Positive control
Ti6Al4V-Cu	polar	-	4h	RTG	100.0	52.5	74.0	38.8	100.0
				MF(× 10^-6)	141.2	149.4	98.6	122.3	515.3
			24h	RTG	100.0	78.0	63.5	96.0	44.7
				MF(× 10^-6)	174.5	157.5	104.1	100.9	470.7
		+	4h	RTG	100.0	64.8	65.6	71.1	55.5
				MF(× 10^-6)	156.0	132.5	129.2	147.1	411.4
	Non-polar	-	4h	RTG	100.0	55.8	58.8	48.5	39.4
				MF(× 10^-6)	129.5	132.7	89.1	100.9	529.4
			24h	RTG	100.0	108.3	113.5	104.9	42.4
				MF(× 10^-6)	117.0	165.4	123.1	176.6	435.1
		+	4h	RTG	100.0	46.5	35.7	122.3	36.5
				MF(× 10^-6)	131.2	127.3	162.0	83.0	378.3
Ti-Cu	Polar	-	4h	RTG	100.0	82.1	87.5	68.3	44.4
				MF(× 10^-6)	151.4	186.4	233.5	249.3	532.1
			24h	RTG	100.0	96.2	69.0	89.0	30.6
				MF(× 10^-6)	102.7	161.3	139.7	119.5	452.6
		+	4h	RTG	100.0	57.0	59.3	34.8	35.6
				MF(× 10^-6)	150.5	192.1	119.1	209.7	622.8
	Non-polar	-	4h	RTG	100.0	87.4	175.8	152.1	60.8
				MF(× 10^-6)	130.3	230.0	193.1	160.0	473.9
			24h	RTG	100.0	193.9	188.4	171.0	85.9
				MF(× 10^-6)	121.5	167.7	135.0	137.0	465.2
		+	4h	RTG	100.0	107.0	178.0	179.0	79.4
				MF(× 10^-6)	170.0	245.8	180.9	160.0	565.4

Sample	Extract Liquid	S9	Time point	Index	Negative control	25%	50%	100%	Positive control
Ti6Al4V-Cu	polar	-	4h	RTG	100.0	52.5	74.0	38.8	100.0
				MF(× 10^-6)	141.2	149.4	98.6	122.3	515.3
			24h	RTG	100.0	78.0	63.5	96.0	44.7
				MF(× 10^-6)	174.5	157.5	104.1	100.9	470.7
		+	4h	RTG	100.0	64.8	65.6	71.1	55.5
				MF(× 10^-6)	156.0	132.5	129.2	147.1	411.4
	Non-polar	-	4h	RTG	100.0	55.8	58.8	48.5	39.4
				MF(× 10^-6)	129.5	132.7	89.1	100.9	529.4
			24h	RTG	100.0	108.3	113.5	104.9	42.4
				MF(× 10^-6)	117.0	165.4	123.1	176.6	435.1
		+	4h	RTG	100.0	46.5	35.7	122.3	36.5
				MF(× 10^-6)	131.2	127.3	162.0	83.0	378.3
Ti-Cu	Polar	-	4h	RTG	100.0	82.1	87.5	68.3	44.4
				MF(× 10^-6)	151.4	186.4	233.5	249.3	532.1
			24h	RTG	100.0	96.2	69.0	89.0	30.6
				MF(× 10^-6)	102.7	161.3	139.7	119.5	452.6
		+	4h	RTG	100.0	57.0	59.3	34.8	35.6
				MF(× 10^-6)	150.5	192.1	119.1	209.7	622.8
	Non-polar	-	4h	RTG	100.0	87.4	175.8	152.1	60.8
				MF(× 10^-6)	130.3	230.0	193.1	160.0	473.9
			24h	RTG	100.0	193.9	188.4	171.0	85.9
				MF(× 10^-6)	121.5	167.7	135.0	137.0	465.2
		+	4h	RTG	100.0	107.0	178.0	179.0	79.4
				MF(× 10^-6)	170.0	245.8	180.9	160.0	565.4

Sample	Extract Liquid	S9	Time point	Negative control	25%	50%	100%	Positive control
Ti6Al4V-Cu	polar	-	4h	0%	0%	0.5%	0	21.5%
			24h	-	0%	0%	0%	-
		+	4h	0%	0%	0%	0%	18%
	Non-polar	-	4h	0%	0%	0%	0%	20.5%
			24h	-	0%	0%	0%	-
		+	4h	0%	0%	0%	0%	18.5%
Ti-Cu	Polar	-	4h	0%	0%	0%	0%	0%
			24h	-	0%	0%	0%	-
		+	4h	0%	0%	0%	0%	0%
	Non-polar	-	4h	0%	0%	0%	0%	20.5%
			24h	-	0%	0%	0%	-
		+	4h	0%	0%	0%	0%	19.5%