A Comprehensive Study on Antibacterial Properties of Mg-HA Nanocomposites for Bone Implants

Appu C. Kurian^1,3, Russel Malkiya Rasalin Prince² , Sreedharan Christopher Ezhil Singh³ and Govind Manoj³
1. Research Scholar,Division of Mechanical Engineering, Karunya Institute of Technology and Sciences, Coimbatore, Tamil Nadu, India
2. Assistant Professor, Division of Mechanical Engineering, Karunya Institute of Technology and Sciences, Coimbatore, Tamil Nadu, India
3. Department of Mechanical Engineering, Vimal Jyothi Engineering College, Kannur, Kerala, India
Correspondence to: Appu C. Kurian, appuckurian@gmail.com

DOI:  https://doi.org/10.70389/PJS.100237

Additional information

• Ethical approval: N/a
• Consent: N/a
• Funding: No industry funding
• Conflicts of interest: N/a
• Author contribution: Appu C. Kurian, R. Malkiya Raslin Prince, S. Christopher Ezhil Singh and Govind Manoj– Conceptualization, Writing – original draft, review and editing.
• Guarantor: Appu C. Kurian
• Provenance and peer-review: Unsolicited and externally peer-reviewed
• Data availability statement: N/a

Keywords: Magnesium-doped hydroxyapatite, Microwave-assisted wet chemical synthesis, Antibacterial bone implant nanocomposites, Biofilm inhibi-tion mechanisms, Mg-HA nanofiber characterization.

Peer Review
Received: 14 August 2025
Last revised: 17 December 2025
Accepted: 17 December 2025
Version accepted: 4
Published: 31 January 2026

Plain Language Summary Infographic

Introduction

A specific area of emphasis in biomaterial research is on bone regeneration because of the prevalence of bone injuries and diseases.^1,2 Traditionally, the treatment includes the use of autografts, but the donor site morbidity,³ besides the limitations due to a scarcity of usable tissue, severely restricts this mode of therapy. Therefore, synthetic alternatives that mimic the characteristics of natural bone have attracted considerable research interest. Among these, hydroxyapatite (HA), which is a natural mineral form of calcium apatite, has emerged as prominent material due to its biocompatibility and bioactivity.^1,3 HA closely resembles the mineral component of bone, hence it can be considered as an ideal candidate for bone grafting and tissue engineering applications.^4,5

However, pure HA lacks mechanical strength and cannot avoid bacterial colonization⁶ that leads to infections associated with implantation.⁴ In light of such challenges, researchers have examined the doping of HA with various metal ions, among which magnesium is very promising.⁷ Magnesium is one of the most important elements involved in bone metabolism; hence, it takes a vital role in osteogenesis and maintains healthy bones.⁸ Incorporation of Mg in the HA structure enhances not only its mechanical properties but also its biological performance, such as increased osteoblastic activity and enhanced biocompatibility.²

Recent studies have demonstrated that magnesium-doped hydroxyapatite (Mg-HA) nanofibers possess superior properties compared to their undoped counterparts.^4,9 Mg-HA nanofibers can be prepared by different methods, e.g., microwave-assisted hydrothermal treatments, where shape and crystallinity of the resulting nanofibers can be well controlled.⁴ Different characterization methods such as X-ray diffraction (XRD), SEM, and Fourier transform infrared spectroscopy (FTIR) have been employed to analyze successful doping of magnesium into Mg-HA nanofibers and structural features of Mg-HA nanofibers.^1,2,10 Mg-HA nanofibers also possess excellent antimicrobial activity. It has been reported that the addition of magnesium ions enhances not only the antibacterial activity of HA but also reduce the opportunistic pathogens, such as Staphylococcus aureus and Escherichia coli.¹¹ These antimicrobial actions can help inhibit infections that may develop with bone implants, thereby avoiding more severe complications and healthcare costs.⁶ The mechanisms underlying the antimicrobial activity of Mg-HA are believed to involve the release of Mg ions, which lyse bacterial cell membranes and induce oxidative stress.^10,12

Besides their antimicrobial action, Mg-HA nanofibers were found to be highly biocompatible In-vitro.^8,9 Cell viability assays analysis showed that Mg-HA nanofibers enhance the proliferation of fibroblasts and osteoblasts, which makes them promising for bone regeneration.^7,13 Most importantly, cytotoxicity of Mg-HA has been found to be low, which further confirms its potential for application in the biomedical field.^5,14 This review aims to present a full picture of the current research situation in the case of Mg-HA nanofibers,³ such as their synthesis, characterization, biocompatibility, antimicrobial activity, and uses in bone regeneration. With the focus put on the developments in this field, it directs attention towards the significance of Mg-HA composite, as a promising biomaterial for future clinical uses.^15,16

Synthesis Methods

There are several routes to synthesize Mg-HA, and each of these has merits in scalability as well as in material properties. Four of the most commonly used techniques are the Microwave-Assisted Wet Chemical Method, Sol-Gel Method, Hydrothermal Synthesis, and Precipitation Method.

Microwave-Assisted Wet Chemical Method: Among the various reported methods, the microwave-assisted wet chemical approach is frequently cited for its accuracy and efficiency in synthesizing Mg-HA. The process starts with dissolving calcium hydroxide (Ca(OH)₂) in distilled water to produce a homogeneous suspension. Ammonium dihydrogen phosphate (NH₄H₂PO₄) is added gradually at a pH of 10, adjusted by adding ammonium hydroxide. Magnesium is added to the HA structure by adding magnesium oxide (MgO), followed by microwave irradiation that increases nucleation and crystallization processes, reducing reaction times by a significant margin compared to conventional processes. During the synthesis, the precipitate is washed, filtered, dried, and calcinated at 900°C to enhance crystallinity (Figure 1). The major advantages of this method are a rapid reaction time, be energy efficient, have uniform magnesium distribution, and reduce particle agglomeration.¹

Sol-Gel Process: The Sol-Gel process (Figure 2) offers a route to good microstructure and compositional control. Phosphoric acid, calcium nitrate, and magnesium nitrate are dissolved in ethanol to create the precursor solution. This is stirred to create a gel-like structure, which is aged for 24–48 hours. After gelation, the material is low-temperature dried (~100°C) and calcined at 600–900°C to enhance crystallinity and burn out organic residues. This route offers high purity homogeneity, and the ability to control particle size and morphology, and hence is extremely suitable to the production of high-quality Mg-HA.

Hydrothermal Synthesis: Hydrothermal synthesis is the method of synthesis of crystalline nanocomposites at high pressure and temperature. Calcium, magnesium, and phosphate aqueous precursors are combined and charged into a sealed autoclave (Figure 3). The solution is heated at 180–200°C for a few hours, and it is possible to synthesize high-quality material. The product is filtered, washed, and dried after the reaction is over. Hydrothermal synthesis is also renowned for the synthesis of highly crystalline, phase-pure material with few impurities and reproducibility.

Precipitation Method: This method is one of the easiest and cheapest approaches for large-scale production. Calcium and magnesium salts, for example, are calcium nitrate (Ca(NO₃)₂) and magnesium nitrate (Mg(NO₃)₂), which are dissolved in distilled water. Precursors of phosphate are added dropwise while continuously stirring to allow for precipitation. Ammonium hydroxide is used to maintain a pH of 9–10, and the resulting precipitate is washed, dried, and calcined. This method is scalable and relatively straightforward, thus being suitable for mass production of Mg-HA.¹ The method referred in this study is Microwave-Assisted Wet Chemical Method due to its accuracy and efficiency in synthesizing Mg-HA.20

Review Methodology

This review was conducted following the PRISMA 2020 (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines to ensure transparency and reproducibility.

Research Question: Which are the synthesis pathways, antimicrobial effects, and biocompatibility of Mg-HA nanocomposites that have been developed as bone implants?

Database Sources: Electronic searches were conducted in Scopus, PubMed, ScienceDirect, SpringerLink, and Google Scholar databases.

Search Strategy and Strings: The following combinations of keywords were used:

(“Magnesium doped hydroxyapatite” OR “Mg–HA”) AND (“antibacterial” OR “antimicrobial”) AND (“biocompatibility” OR “cytotoxicity”) AND (“synthesis” OR “nanocomposite” OR “bone implant”).

Search Period: Studies published between January 2010 and October 2024 were considered.

• Inclusion Criteria: Peer-reviewed journal articles and reviews in English. Research that has described synthesis procedures, antibacterial or cytotoxicity of Mg-HA composites. In vitro or in vivo investigations on biomedical applications.
• Exclusion Criteria: Articles not related to Mg-HA (e.g., other dopants or alloys).
• Non-English or unpublished materials. Conference abstracts without experimental or analytical data.

Study Selection Flow: An initial search yielded 312 records, of which 78 duplicates were removed. Titles and abstracts of 234 studies were screened; 61 full-text papers were assessed for eligibility. Finally, 21 articles meeting all inclusion criteria were included in the qualitative synthesis (Figure 4).

Risk-of-Bias Assessment: A modified version of the Joanna Briggs Institute checklist on experimental and observational studies was used to assess risk of bias in individual studies, and it addressed the questions of completeness of data, sample size, and reproducibility.

Methodology

This review compiles data from studies that analyzed antibacterial performance of Mg-HA specimens prepared through various synthesis routes. The methodology outlines (Figure 5) the preparation of HA particles for various nanocomposite samples, which are to be applied in the human body. Since the composites are designed for implantation in the human body, antibacterial studies are vital to assess their interactions during the degradation process.^1,4 Reported studies have examined various weight percentages of Mg incorporation to identify optimal compositions.

Characterization Techniques

X-ray Diffraction (XRD): XRD is employed to determine the crystalline structure and phase purity of the synthesized Mg-HA. It provides information on the crystallinity, phase composition, and average crystallite size. The diffraction peaks from the XRD patterns are analyzed by comparing with standard reference patterns, such as JCPDS card no: 09-0432 for HA. Absence of second phases indicates the material’s purity during formation, while sharp peaks indicate high crystallinity.¹

Fourier Transform Infrared Spectroscopy (FTIR)

FTIR spectroscopy was employed in identifying the functional groups of Mg-HA and also in ensuring the incorporation of magnesium into the HA structure. The FTIR spectra of the samples have different absorption bands for phosphate groups (ν1 at 665 cm⁻¹) and hydroxyl (ν3 at 3574 cm⁻¹). Appearance of these bands tells us that HA has been successfully prepared. Moreover, any shift or changes in intensity or peak position may give indications about the magnesium ions and the HA matrix interaction.^1,3

Antibacterial Activity of Magnesium-Substituted Hydroxyapatite

Mode of Antibacterial Activity

Mechanisms of Action: Disruption of Bacterial Cell Walls: The incorporation of magnesium ions (Mg²⁺) into the hydroxyapatite lattice significantly enhances the antibacterial properties of HA. Upon penetration of Mg²⁺ ions into the bacterial cell membrane, there is disturbance of the ionic balance and metabolic processes in the cell. This disruption leads to the structural integrity of the cell wall, causing leakage of intracellular components and ultimately resulting in cell lysis and death. This mechanism is highly potent towards Gram-positive organisms, for instance, Staphylococcus aureus which has a significant peptidoglycan layer with considerable ionic liability. Alanis Gómez et al., has described that Mg²⁺ can cause an extreme enhancement of membrane permeability that allows the accidental entry of foreign substances into the cells while allowing essential cellular contents to leak out.

Induction of Oxidative Stress: Mg²⁺ ion not only causes membrane damage but also has the potential to induce oxidative stress in bacterial cells. It is brought on by the production of highly reactive molecules known as reactive oxygen species (ROS), which can potentially cause damage to many cellular components such as proteins, lipids, and nucleic acids. Cellular damage and death are results of oxidative stress caused by ROS. The interaction of Mg²⁺ ions with cellular constituents results in the production of free radicals, which initiate peroxidation of lipids and protein denaturation reactions.⁴ Mg-HA is an extremely potent antibacterial compound because of its dual mechanism of action: membrane disruption and oxidative stress generation.

Biofilm Inhibition

Quorum Sensing Interference: Biofilm formation is of critical significance in the determination of the chronic nature of bacterial infections, particularly in the context of medical implants. Biofilm-encased bacteria are shielded from the immune system and antibiotics by a matrix of an extracellular polymeric substance (EPS). The inhibition of bacterial quorum sensing, a key chemical signaling pathway involved in microbial communication and biofilm establishment, enables the substance to suppress biofilm formation. Biofilm formation in the initial stages is prevented in laboratory experiments when Mg²⁺ ions are present since they prevent bacterial cell adhesion onto surfaces.³ This interference of quorum sensing not only inhibits the establishment of new biofilm but also enables the removal of pre-existing bacteria that form biofilms, thereby the effectiveness of Magnesium-doped HA in clinical applications.

Inhibition of EPS Production: Mg-HA not only disrupts bacterial communication but also inhibits the production of the EPS matrix itself. This lowers the chances of implant failure caused by bacterial colonisation by stopping biofilm maturation and having a cleaner surface. Mg-HA is a good material for the preparation of antimicrobial coatings for medical devices because of its inhibitory activity against biofilm growth. Another evidence of Mg-HA’s excellent inhibitory activity as a biofilm inhibitor is that it greatly decreases the expression of bacterial genes responsible for extracellular EPS production.⁴

Testing Methods for Antibacterial Properties

Disk Diffusion Method

The disc diffusion technique is a popular method for finding the antibacterial activity of different materials. In accordance with this technique, nutrient agar plates are carefully prepared, and then inoculated with standardized suspensions of bacteria, typically at a concentration of 1 × 10⁶ CFU/mL, uniformly distributed on the surface of the agar by the aseptic technique. Sterilized filter paper discs with different concentrations of Mg-HA (e.g., 50 µL, 100 µL, and 150 µL) are put on the agar surface along with control discs infused with a standard antibacterial drug (e.g., Diclofenac sodium) and plain discs for comparative study.⁴

Incubation and Measurement: Culture plates were incubated at 37°C, exposed to optimal growth conditions for 24 hours. The diameters of the inhibition zones around the discs are measured with a millimeter ruler after incubation time. A measure of the antibacterial activity of the Mg-HA samples is the inhibition zone measurement; the larger the zone, the higher the antibacterial activity. The technique is used in microbiology and allows simple visual estimation of antibacterial activity.³

Minimum Inhibitory Concentration (MIC)

Minimum inhibitory concentration (MIC) is determined by the minimum concentration of Mg-HA that prevents visible growth of bacteria. The same amount of bacterial suspension is added to all Mg-HA dilutions prepared in an aqueous culture medium. Bacterial growth is checked visually for turbidity or is measured quantitatively by a spectrophotometric determination of the 600 nm optical density following incubation at 37°C.^2,17 The MIC is the lowest concentration of Mg-HA at which neither the lowest absorbance nor any discernible growth occurs. The process may be compared with other antibacterial substances and provides valuable information on the inhibitory activity of Mg-HA for bacterial growth at varying doses. The determined MIC values can help choose the right concentrations for additional in vivo research and clinical uses.³

Comparitive Analysis

Performance Relative to Reference Antibacterial Agents

As per the outcome of the MIC and disc diffusion tests, Mg-HA is more antibacterial than pure HA. Specifically, Mg-HA showed much higher inhibition zones against E. coli and S. aureus in the disc diffusion tests, demonstrating its higher bactericidal activity.¹ Mg-HA showed inhibition zones of 6.7 mm against E. coli and 6.4 mm against S. aureus at 150 µL concentration, whereas the HA samples showed smaller inhibition zones.

MIC Values: Additional proof of the high potential of the Mg-HA composite to inhibit growth of bacterial cells at lower doses was provided by the observation that it had lower MIC levels than HA. The increased antibacterial activity is due to the presence of magnesium ions, which destabilize not only the bacterial membrane integrity but also create oxidative stress and biofilm inhibition.³ This comparative study indicates Mg-HA as a candidate alternative that can potentially outperform conventional antibiotics in preventing medical implant infections.

Mechanistic Insights

Increased Antibacterial Activity: The following are the reasons for the increased antibacterial activity of Mg-HA:

• Membrane Disruption: Mg²⁺ ions cause generalized disruption to bacterial membranes, leading to cell lysis.
• Increased ROS Production: ROS generation is the cause of oxidative stress, which further kills bacteria.
• Biofilm Prevention: Mg-HA effectively prevents biofilm formation by disrupting bacterial communication and inhibiting EPS production.¹

Comparative Analysis with Standards: Table 1 provides a comparative summary of Mg-HA and related materials reported in the literature alongside the present study. Unlike pure HA and other metal-doped variants (e.g., ZnO–HA, Cu–HA), reported Mg-HA nanocomposites have exhibited larger inhibition zones than pure HA and other metal-doped variants both E. coli and S. aureus (6.7 and 6.4 mm, respectively) while maintaining low cytotoxicity. This balance of strong antibacterial performance and excellent biocompatibility highlights Mg-HA as a more reliable candidate for orthopedic and dental implant coatings compared to other reported biomaterials. Compared to traditional antibacterial agents, Mg-HA provided similar or even superior results, particularly at higher concentrations. Magnesium addition not only enhances antibacterial efficacy but also maintains biocompatibility, evidenced through hemolysis and cytotoxicity tests.^3,4 The dual benefit makes Mg-HA a viable option for clinical application in bone regeneration and infection resistance.

Table 1: Comparison of various methods in Mg-HA preparations.
Author(s)	Material Used	Fabrication Process	Key Findings/ Outcomes	Applications
Jenifer et al.1	Mg-HA	Wet chemical synthesis	Good antibacterial and haemolytic properties. Cytotoxicity assays describe the MTT and AlamarBlue (fibroblast proliferation; significance reported).	Bone tissue engineering
Chen et al.6	Magnesium Alloys	Review (casting, extrusion, etc.)	Biodegradable, good mechanical and corrosion resistance. Biofilm formation assays (OD₄₉₀ nm), concentration series (mg/mL).	Biodegradable implants
Catarina et al.4	HA + MgO	Mixing/sintering	Antibacterial bone substitute, infection prevention. Review compiles larger/smaller zones vs pure HA.	Dental and orthopaedic applications
Alanis Gómez et al.2	Mg-HA Nanofibers	Electrospinning	Antimicrobial, cytocompatibility. Reported “strong antibacterial effects”.	Biomedical applications
Grenho et al.21	Nano HA–ZnO	Chemical synthesis	Strong antibacterial activity. Inhibition at all tested concentrations from 5 mg/mL down to 0.009 mg/mL.	Bone and dental implants
Hudson et al.11	Bone Matrix & Biomaterials	Biological assay	S. aureus adheres strongly to bone surfaces.	Infection mechanism studies
Shemesh et al.14	Streptococcus mutans	Biofilm formation on surfaces	Genetic adaptation varies by surface type.	Dental material improvement
Sawai22	MgO, ZnO, CaO	Conductimetric assay	MgO shows good antibacterial activity. Biofilm inhibition starting at 2.5 mg/mL.	Dental, antimicrobial agents
Fang et al.6	Various inorganic agents	Susceptibility tests	Effective antibacterial against oral bacteria.	Oral infection control
Shanmugam and Gopal23	Cu-substituted HA/Fluorapatite	Solid-state reaction	Strong antimicrobial properties. Increased fibroblast proliferation with 2% and 5% HA-Mg vs controls.	Orthopaedic/dental coatings
Lemire et al.13	Metals (e.g. Ag, Cu, Zn)	Review	Detailed antimicrobial mechanisms of metals.	Medical and environmental uses
Azam et al.10	Metal oxide nanoparticles	Comparative study	Effective against both Gram+ and Gram− bacteria.	Nanomedicine, disinfection
Shafiee et al.15 Kumar et al19	Mg/HA Bio-composite	Stir casting	Improved mechanical and bio-properties.	Orthopaedic/dental biomaterials
Rahmany-Gorji et al.24	ZX51/Al₂O₃ Mg Composite	Stir casting	Enhanced microstructure, improved mechanical properties. Review states low cytotoxicity and enhanced cell growth for Mg-HA materials; specific numeric cell viability percentages are reported.	Biomedical & structural uses
Prasadh et al.18	Hollow Silica-Reinforced Mg	Powder metallurgy + modelling	Enhanced strength & biocompatibility.	Mandibular reconstruction
Razal Rose and Aatthisugan16	Magnesium Metal Composite	Stir casting	Strong mechanical properties; structural integrity.	Biomedical and automotive applications

Scope and Limitation

Although there is substantial evidence to support the antibacterial and osteogenic potential of Mg-HA, there are several factors that cause heterogeneity in the available results. The crystallinity, ion release, and surface area are greatly affected by the differences in the particle size, the level of Mg²⁺ substitution, and the synthesis pathways, thereby influencing the antibacterial and biocompatibility results. Besides this, assay conditions (bacterial strain, culture medium, inoculum density and incubation period) among studies are rather diverse which limits direct comparison of inhibition-zone and MIC data. Other methodological biases are the usage of small samples, non-standardized antibacterial procedures, or inadequate reporting of statistics. The only reports that use such reference standards as CLSI or ISO antimicrobial testing methods may influence reproducibility.

At the translational level, the Mg-HA systems have to match the antibacterial effectiveness with corrosion control and the release of Mg-ion because too much corrosion might increase the local alkalinity and surpass the cytotoxicity levels. It is exploring using surface coating, polymer encasement or alloying as a method of controlling the kinetics of corrosion. Moreover, the majority of the existing data are based on the in-vitro assessments; there is little in-vivo data available on the long-term osseointegration, inflammatory reactions, and the controlled Mg-ion dissolution. Future research must thus incorporate standardized testing protocol, quantitative corrosion evaluation as well as preclinical in-vivo validation to enhance translational relevance.

Conclusion

Mg-HA nanofibers, which are formed by a variety of procedures like microwave-assisted hydrothermal processing, are an important improvement of biomaterials. The presence of magnesium also improves mechanical and biological characteristics, giving them effective antimicrobial efficiency against most implant-associated pathogens and demonstrated ability to induce bone regeneration. This bi-functionality along with proven biocompatibility and low cytotoxicity, makes Mg-HA nanofibers a very promising alternative to existing orthopedic and dental bone graft material. The next wave of research should be conducted on the clarification of the underlying mechanism of its biological property and its performance in living models to further confirm its clinical potential in bone regeneration and infection prevention.

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Cite this article as:
Kurian AC, Prince RMR, Singh SCE and Manoj G. A Comprehensive Study on Antibacterial Properties of Mg-HA Nanocomposites for Bone Implants. Premier Journal of Science 2025;15:100237

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