Fabrication and in vitro Biocompatibility Evaluation of BA + Alg@Ca Composite Coatings on Biodegradable Pure Mg

doi:10.11900/0412.1961.2024.00181

Acta Metall Sin

2026, Vol. 62

Issue (4): 669-684 DOI: 10.11900/0412.1961.2024.00181

Research paper

Current Issue | Archive | Adv Search

Fabrication and in vitro Biocompatibility Evaluation of BA + Alg@Ca Composite Coatings on Biodegradable Pure Mg

LIANG Tao¹^,², DU Yunbo¹, CHEN Xiehui¹, PAN Haobo²^,³(

)

^1.Shenzhen Longhua Central Hospital, Shenzhen 518110, China
^2.Shenzhen Key Laboratory of Marine Biomedical Materials, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
^3.Shenzhen Healthemes Biotechnology Co. Ltd. , Shenzhen 518044, China

Cite this article:

LIANG Tao, DU Yunbo, CHEN Xiehui, PAN Haobo. Fabrication and in vitro Biocompatibility Evaluation of BA + Alg@Ca Composite Coatings on Biodegradable Pure Mg. Acta Metall Sin, 2026, 62(4): 669-684.

Download:

HTML

PDF(4490KB)
Export: BibTeX | EndNote (RIS)

Abstract

Infected bone defect is one of the most common and serious diseases in orthopedic surgery. Hence, it is important to design bone tissue engineering biomaterials with both antibacterial and osteogenic properties for the repair of infected bone defects. Accordingly, in this study, BA + Alg@Ca composite coatings (BA represents H₃BO₃ hydrothermal treatment, Alg@Ca represents CaG-incorporated alginate) composed of an inner layer (Mg(OH)₂), a middle layer (MgB₂O(OH)₆), and an outer layer (Alg@Ca) were constructed on pure Mg by hydrothermal treatment and dip coating, and the formation mechanism and in vitro biocompatibility were systematically investigated. BA + Alg@Ca coatings dramatically improved the degradation resistance of pure Mg, revealing a lower pH value and released Mg²⁺ concentration in the immersion test as well as nobler open circuit potential, larger impedance modulus, and lower corrosion current density in the electrochemical evaluation. In vitro antibacterial examination showed that BA + Alg@Ca coatings effectively inhibited the growth of gram-positive and gram-negative bacteria due to the synergetic effects of [B(OH)₄]^- and alginate. BA + Alg@Ca coatings also demonstrated good cytocompatibility with normal cell viability and proliferation. Moreover, these coatings could induce macrophage polarization to the M2 phenotype (anti-inflammatory), suggesting that they resulted in favorably selective cell adhesion and antibacterial performance. Overall, the BA + Alg@Ca coating-modified pure Mg can potentially be used for the repair of infected bone defects in the clinic.

Key words: pure Mg H₃BO₃ sodium alginate antibacterial property biocompatibility

Received: 28 May 2024

ZTFLH:

TG146.2

Fund: National Natural Science Foundation of China(32161160327);Guangdong Basic and Applied Basic Research Foundation of China(2022A1515110901);Shenzhen Medical Research Fund(B2302031)

Corresponding Authors: PAN Haobo, professor, Tel: 15986769058, E-mail: hb.pan@siat.ac.cn

	Service

	E-mail this article
	Add to citation manager
	E-mail Alert
	RSS
	（）
	Articles by authors
	LIANG Tao
	DU Yunbo
	CHEN Xiehui
	PAN Haobo

URL:

https://www.ams.org.cn/EN/10.11900/0412.1961.2024.00181 OR https://www.ams.org.cn/EN/Y2026/V62/I4/669

Table 1 Primer pairs of the related genes used in the quantitative real-time polymerase chain reaction (qRT-PCR) assay

Fig.1 Surface SEM images of BA coatings prepared by treating pure Mg with various concentrations of H₃BO₃ solutions (BA—H₃BO₃ hydrothermal treatment)
(a) untreated (b) 0.05 mol/L (BA(0.05)) (c) 0.10 mol/L (BA(0.10)) (d) 0.20 mol/L (BA(0.20))

Fig.2 Open circuit potential (OCP) curves (a), Nyquist plots (b), and potential dynamic curves (c) of BA samples (SCE—saturated calomel electrode, Z_Re—real part of impedance, Z_Im—imaginary part of impedance)

Fig.3 Antibacterial properties of control, untreated, and BA samples against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Acinetobacterbaumannii (A. baumannii) (Ti6Al4V titanium alloy was used as control sample, the same below)

Fig.4 Characterizations of surface and cross-sectional properties
(a) water contact angles of untreated pure Mg, BA, BA + Alg, and BA + Alg@Ca samples
(b) surface morphologies of untreated pure Mg, BA, BA + Alg, and BA + Alg@Ca samples (Insets show the lower magnification images)
(c) SEM image and corresponding EDS mappings of cross-section of BA + Alg@Ca sample

Fig.5 XRD patterns of (a), attenuated total reflection-Fourier transform infrared (ATR-FTIR) patterns (b), and XPS (c) of different samples; and high resolution XPS of B (d), Mg (e), and Ca (f)

Fig.6 Immersion results of different samples in phosphate buffer saline (PBS) solution
(a) pH value (b) Mg²⁺ concentration (c) [B(OH)₄]^- concentration (d) Ca²⁺ concentration

Fig.7 Electrochemical results of untreated pure Mg, BA, BA + Alg, and BA + Alg@Ca samples in PBS solution (|Z|—modulus of impedance)
(a) OCP curves
(b) Bode plots
(c) potentiodynamic polarization curves

Fig.8 Qualitative (a-c) and quantitative (d-f) results of antioxidant abilities of samples in PBS solution (CAT—catalase, SOD—superoxide dismutase, DPPH $⋅$ —2, 2-diphenyl-1-picrylhydrazyl)
(a, d) CAT-like activity (b, e) SOD-like activity (c, f) DPPH

⋅

free radical scavenging activity

Fig.9 Cytocompatibility results of different samples (OD₄₅₀ in Fig.9a represents optical density at the wavelength of 450 nm; red and blue areas in Fig.9b represent cytoskeleton and cell nuclei, respectively)
(a) cell counting kit-8 (CCK8) assay of extracts
(b) fluorescent images of cells co-cultured with sample extracts for 7 d

Fig.10 In vitro antibacterial properties of untreated pure Mg, BA, BA + Alg, and BA + Alg@Ca samples
(a) representative photos of bacterial colonies
(b) antibacterial rate against S. aureus and Staphylococcus epidermidis (S. epidermidis)
(c) antibacterial rate against E. coli and A. baumannii

Fig.11 Immunomodulatory properties of untreated pure Mg, BA, BA + Alg, and BA + Alg@Ca samples on RAW264.7 macrophage cells
(a, b) enzyme-linked immunosorbent assay (ELISA) results of inflammatory factors (a) and anti-inflammatory factors (b)
(c, d) relative mRNA expression of immune related genes of inflammatory genes (c) and anti-inflammatory genes (d)

[1]	Vallet-Regí M, Lozano D, González B, et al. Biomaterials against bone infection [J]. Adv. Healthc. Mater., 2020, 9: e2000310
[2]	Thomas M V, Puleo D A. Infection, inflammation, and bone regeneration: A paradoxical relationship [J]. J. Dent. Res., 2011, 90: 1052
[3]	Qiu Y, Zhang N, An Y H, et al. Biomaterial strategies to reduce implant-associated infections [J]. Int. J. Artif. Organs., 2007, 30: 828
[4]	Lin X, Ge J, Wu S L, et al. Advances in metallic biomaterials with both osteogenic and anti-infection properties [J]. Acta Metall. Sin., 2017, 53: 1284
	林潇, 葛隽, 吴水林等. 兼具成骨和抗感染性能的医用金属材料研究进展 [J]. 金属学报, 2017, 53: 1284
[5]	Han H S, Loffredo S, Jun I, et al. Current status and outlook on the clinical translation of biodegradable metals [J]. Mater. Today, 2019, 23: 57
[6]	Li N, Zheng Y F. Novel magnesium alloys developed for biomedical application: A review [J]. J. Mater. Sci. Technol., 2013, 29: 489
[7]	Dong J H, Tan L L, Yang K. Research of biodegradable Mg-based metals as bone graft substitutes [J]. Acta Metall. Sin., 2017, 53: 1197
	东家慧, 谭丽丽, 杨柯. 可降解镁基金属骨缺损修复材料的研究探索 [J]. 金属学报, 2017, 53: 1197
[8]	Han H S, Jun I, Seok H K, et al. Biodegradable magnesium alloys promote angio-osteogenesis to enhance bone repair [J]. Adv. Sci., 2020, 7: 2000800
[9]	Skalny A V, Aschner M, Silina E V, et al. The role of trace elements and minerals in osteoporosis: A review of epidemiological and laboratory findings [J]. Biomolecules, 2023, 13: 1006
[10]	Cheng Q H, Liu J, Zhang W H, et al. Research progress of magnesium in the treatment of osteoporosis [J]. J. Ningxia Med. Univ., 2022, 44: 957
	程晴灏, 刘杰, 张文辉等. 镁与骨质疏松治疗的研究进展 [J]. 宁夏医科大学学报, 2022, 44: 957
[11]	Feng H Q, Wang G M, Jin W H, et al. Systematic study of inherent antibacterial properties of magnesium-based biomaterials [J]. ACS Appl. Mater. Interfaces, 2016, 8: 9662
[12]	Robinson D A, Griffith R W, Shechtman D, et al. In vitro antibacterial properties of magnesium metal against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus [J]. Acta Biomater., 2010, 6: 1869
[13]	Qin H, Zhao Y C, Cheng M Q, et al. Anti-biofilm properties of magnesium metal via alkaline pH [J]. RSC Adv., 2015, 5: 21434
[14]	Hou P, Zhao C L, Cheng P F, et al. Reduced antibacterial property of metallic magnesium in vivo [J]. Biomed. Mater., 2016, 12: 015010
[15]	Liu Y, Zheng Y F, Chen X H, et al. Fundamental theory of biodegradable metals—Definition, criteria, and design [J]. Adv. Funct. Mater., 2019, 29: 1805402
[16]	Zhao Y, Zeng L L, Liang T. Development in biocompatibility of biodegradable magnesium-based metals [J]. Acta Metall. Sin., 2017, 53: 1181
	赵颖, 曾利兰, 梁涛. 可降解镁基金属的生物相容性研究进展 [J]. 金属学报, 2017, 53: 1181
[17]	Esmaily M, Svensson J E, Fajardo S, et al. Fundamentals and advances in magnesium alloy corrosion [J]. Prog. Mater. Sci., 2017, 89: 92
[18]	Cui Z D, Zhu J M, Jiang H, et al. Research progress of the surface modification of titanium and titanium alloys for biomedical application [J]. Acta Metall. Sin., 2022, 58: 837
	崔振铎, 朱家民, 姜辉等. Ti及钛合金表面改性在生物医用领域的研究进展 [J]. 金属学报, 2022, 58: 837
[19]	Wang G M, Jiang W J, Mo S, et al. Nonleaching antibacterial concept demonstrated by in situ construction of 2D nanoflakes on magnesium [J]. Adv. Sci., 2020, 7: 1902089
[20]	Zhang D D, Cheng S, Tan J, et al. Black Mn-containing layered double hydroxide coated magnesium alloy for osteosarcoma therapy, bacteria killing, and bone regeneration [J]. Bioact. Mater., 2022, 17: 394
[21]	Zhang D, Han Q, Yu K, et al. Antibacterial activities against Porphyromonas gingivalis and biological characteristics of copper-bearing PEO coatings on magnesium [J]. J. Mater. Sci. Technol., 2021, 61: 33
[22]	Song J Q, Jin P L, Li M Q, et al. Antibacterial properties and biocompatibility in vivo and vitro of composite coating of pure magnesium ultrasonic micro-arc oxidation phytic acid copper loaded [J]. J. Mater. Sci.-Mater. Med., 2019, 30: 49
[23]	Abodunrin O D, El Mabrouk K, Bricha M. A review on borate bioactive glasses (BBG): Effect of doping elements, degradation, and applications [J]. J. Mater. Chem., 2023, 11B: 955
[24]	Meng Y, Chen L J, Chen Y, et al. Reactive metal boride nanoparticles trap lipopolysaccharide and peptidoglycan for bacteria-infected wound healing [J]. Nat. Commun., 2022, 13: 7353
[25]	Pizzorno L. Nothing boring about boron [J]. Integr. Med. (Encinitas), 2015, 14: 35
[26]	Lopalco A, Lopedota A A, Laquintana V, et al. Boric acid, a lewis acid with unique and unusual properties: Formulation implications [J]. J. Pharm. Sci., 2020, 109: 2375
[27]	Trippier P C, McGuigan C. Boronic acids in medicinal chemistry: Anticancer, antibacterial and antiviral applications [J]. Med. Chem. Comm., 2010, 1: 183
[28]	Duan H P, Yan C W, Wang F H. Effect of electrolyte additives on performance of plasma electrolytic oxidation films formed on magnesium alloy AZ91D [J]. Electrochim. Acta, 2007, 52: 3785
[29]	LI X Y, Bandyopadhyay P, Guo M, et al. Enhanced gas barrier and anticorrosion performance of boric acid induced cross-linked poly(vinyl alcohol-co-ethylene)/graphene oxide film [J]. Carbon, 2018, 133: 150
[30]	Woo J H, Kim N H, Kim S I, et al. Effects of the addition of boric acid on the physical properties of MXene/polyvinyl alcohol (PVA) nanocomposite [J]. Composites, 2020, 199B: 108205
[31]	Song T, Gao F F, Du X, et al. Removal of boron in aqueous solution by magnesium oxide with the hydration process [J]. Colloid. Surf., 2023, 665A: 131211
[32]	Wang Q Y, Li L P, Tian Y, et al. Shapeable amino-functionalized sodium alginate aerogel for high-performance adsorption of Cr(VI) and Cd(II): Experimental and theoretical investigations [J]. Chem. Eng. J., 2022, 446: 137430
[33]	Yan B R, Hu X M, Zhao Y Y, et al. Research and development of a sodium alginate/calcium ion gel based on in situ cross-linked double-network for controlling spontaneous combustion of coal [J]. Fuel, 2022, 322: 124260
[34]	Wang L, Gao H Y, Song S M, et al. The depressing role of sodium alginate in the flotation of Ca²⁺-activated quartz using fatty acid collector [J]. J. Mol. Liq., 2021, 343: 117618
[35]	Montes L, Gisbert M, Hinojosa I, et al. Impact of drying on the sodium alginate obtained after polyphenols ultrasound-assisted extraction from Ascophyllum nodosum seaweeds [J]. Carbohyd. Polym., 2021, 272: 118455
[36]	Costa-Serge N D M, Lima Gonçalves R G, Ramirez-Ubillus M A, et al. Effect of the interlamellar anion on CuMgFe-LDH in solar photo-Fenton and Fenton-like degradation of the anticancer drug 5-fluorouracil [J]. Appl. Catal., 2022, 315B: 121537
[37]	Wang J X, Zhang G Q, Qiao S, et al. Comparative assessment of formation pathways and adsorption behavior reveals the role of NaOH of MgO-modified diatomite on phosphate recovery [J]. Sci. Total Environ., 2023, 876: 162785
[38]	Tang S X, Yang J Y, Lin L Z, et al. Construction of physically crosslinked chitosan/sodium alginate/calcium ion double-network hydrogel and its application to heavy metal ions removal [J]. Chem. Eng. J., 2020, 393: 124728
[39]	Yu X M, Tan L L, Liu Z Y, et al. Preparation and properties of biological functional magnesium coating on Ti6Al4V substrate [J]. Acta Metall. Sin., 2018, 54: 943
	于晓明, 谭丽丽, 刘宗元等. Ti6Al4V表面生物功能纯Mg薄膜制备及性能研究 [J]. 金属学报, 2018, 54: 943
[40]	Feng M C, Fu Q Y, Li J, et al. Sodium alginate coating on biodegradable high-purity magnesium with a hydroxide/silane transition layer for corrosion retardation [J]. Colloid. Surf., 2022, 642A: 128647
[41]	Zhao K Y, Zhang X X, Wei J F, et al. Calcium alginate hydrogel filtration membrane with excellent anti-fouling property and controlled separation performance [J]. J. Membr. Sci., 2015, 492: 536
[42]	Benslima A, Sellimi S, Hamdi M, et al. The brown seaweed Cystoseira schiffneri as a source of sodium alginate: Chemical and structural characterization, and antioxidant activities [J]. Food Biosci., 2021, 40: 100873

[1]	ZHENG Yufeng, SHEN Yunong. Research Status and Future Directions of Pure Mo and Mo-Based Biodegradable Metals[J]. 金属学报, 2026, 62(5): 905-922.
[2]	LIN Xue, QIAN Junyu, ZHANG Wentai, WANG Peng, WAN Guojiang. Osteogenic and Antibacterial Metal-Polyphenol Drug-Loaded Coating on Biodegradable Zinc for Orthopedic Implants Application[J]. 金属学报, 2024, 60(11): 1545-1558.
[3]	ZENG Yunpeng, YAN Wei, SHI Xianbo, YAN Maocheng, SHAN Yiyin, YANG Ke. Effect of Copper Content on the MIC Resistance in Pipeline Steel[J]. 金属学报, 2024, 60(1): 43-56.
[4]	WANG Luning, YIN Yuxia, SHI Zhangzhi, HAN Qianqian. Research Progress on Biocompatibility Evaluation of Biomedical Degradable Zinc Alloys[J]. 金属学报, 2023, 59(3): 319-334.
[5]	CUI Zhenduo, ZHU Jiamin, JIANG Hui, WU Shuilin, ZHU Shengli. Research Progress of the Surface Modification of Titanium and Titanium Alloys for Biomedical Application[J]. 金属学报, 2022, 58(7): 837-856.
[6]	ZHENG Yufeng, XIA Dandan, SHEN Yunong, LIU Yunsong, XU Yuqian, WEN Peng, TIAN Yun, LAI Yuxiao. Additively Manufactured Biodegrabable Metal Implants[J]. 金属学报, 2021, 57(11): 1499-1520.
[7]	Xiaoming YU, Lili TAN, Zongyuan LIU, Ke YANG, Zhonglin ZHU, Yangde LI. Preparation and Properties of Biological Functional Magnesium Coating on Ti6Al4V Substrate[J]. 金属学报, 2018, 54(6): 943-949.
[8]	Erlin ZHANG, Xiaoyan WANG, Yong HAN. Research Status of Biomedical Porous Ti and Its Alloy in China[J]. 金属学报, 2017, 53(12): 1555-1567.
[9]	Guangyin YUAN, Jialin NIU. Research Progress of Biodegradable Magnesium Alloys for Orthopedic Applications[J]. 金属学报, 2017, 53(10): 1168-1180.
[10]	Lili TAN, Junxiu CHEN, Xiaoming YU, Ke YANG. Recent Advances on Biodegradable MgYREZrMagnesium Alloy[J]. 金属学报, 2017, 53(10): 1207-1214.
[11]	Yufeng ZHENG, Hongtao YANG. Research Progress in Biodegradable Metals forStent Application[J]. 金属学报, 2017, 53(10): 1227-1237.
[12]	Chunyong LIANG, Jingzu HAO, Hongshui WANG, Baoe LI, Dan XIA. Preparation and Research Progress of Contact-Induced Surface of Metal Implants[J]. 金属学报, 2017, 53(10): 1265-1283.
[13]	Luning WANG, Yao MENG, Lijun LIU, Chaofang DONG, Yu YAN. Research Progress on Biodegradable Zinc-Based Biomaterials[J]. 金属学报, 2017, 53(10): 1317-1322.
[14]	Cong PENG, Shuyuan ZHANG, Ling REN, Ke YANG. Effect of Cooling Rate on Microstructure and Properties ofa Cu-Containing Titanium Alloy[J]. 金属学报, 2017, 53(10): 1377-1384.
[15]	Xinyin WANG, Yan XIA, Yaru ZHOU, Linlin NIE, Fahe CAO, Jianqing ZHANG, Chunan CAO. CORROSION BEHAVIOR OF PURE Mg BASED ON GENERATION/COLLECTION AND FEEDBACK MODES OF SCANNING ELECTROCHEMICAL MICROSCOPY[J]. 金属学报, 2015, 51(5): 631-640.

No Suggested Reading articles found!

Viewed

Full text

Abstract

Cited

Shared

Discussed