A multifunctional gallium-mof/hydrogel construct based on tetratopic ligands and pectin: structural optimization and biomedical potential

A novel hydrogel-based material was synthesized using gallium nitrate, a tetratopic pyridine-carboxylate ligand (H₄TBAPy), oxidized pectin, and chitosan (Gallium-MOF/Hydrogel). This composite material incorporates a metal–organic framework (MOF) network within a biopolymeric hydrogel matrix. The structure was characterized via scanning electron microscopy (SEM), Brunauer–Emmett–Teller (BET) surface area analysis, carbon/hydrogen/nitrogen/oxygen elemental analysis (CHNO EA), Fourier-transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), energy-dispersive X-ray (EDX) and EDX mapping, confirming the formation of a nanoscale MOF-hydrogel system with high surface area and uniform morphology. The antimicrobial activity of the material was evaluated against clinically relevant fungal species and Gram-positive and Gram-negative bacterial strains, showing superior minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and minimum bactericidal concentration (MBC) values compared to two standard antibiotics. Furthermore, cytotoxicity assays on against skin (A-431), breast (MCF-7), and bone cancer (MG-63) cancer cells revealed strong anticancer effects, likely due to the bioactive nature of the Ga-MOF core and synergistic interactions with pectin and chitosan. The obtained results highlight the potential of this Ga-based hydrogel as a multifunctional platform for biomedical applications.

1 Introduction

In recent years, the development of advanced materials has become a central focus in addressing challenges in biomedical and sensing technologies. Systems such as hydrogel-based drug delivery platforms and novel functional derivatives have attracted considerable interest due to their biocompatibility, tunable properties, and potential for precise therapeutic interventions. Highlighting the progress in these areas not only emphasizes their scientific significance but also sets the stage for exploring new classes of materials with even greater versatility. Within this broader context, increasing attention has been directed toward metal-organic frameworks (MOFs) as promising candidates for next-generation applications (Wang et al., 2023; Zhong et al., 2024).

MOFs are crystalline porous materials composed of metal ions or clusters coordinated with multidentate organic ligands (Liu et al., 2024). Their high surface area, adjustable pore size, and structural tunability have made them valuable in diverse fields such as gas separation (Felix Sahayaraj et al., 2023), catalysis (Li et al., 2023), biosensing, and drug delivery (Saboorizadeh et al., 2024). In recent years, the emergence of biologically functional MOFs (bio-MOFs), particularly those constructed from biocompatible components, has opened new avenues in biomedical applications, including antimicrobial therapy and targeted cancer treatment (Sadiq et al., 2024).

Among the biocompatible metal ions explored for such applications, gallium (Ga⁺³) has attracted considerable attention due to its ability to mimic ferric ions (Fe⁺³) in biological systems (Liu et al., 2025). This enables gallium to disrupt iron-dependent metabolic processes essential for microbial survival and tumor growth (Truong et al., 2023; Yao et al., 2024). Gallium-based compounds have been clinically used in the treatment of hypercalcemia of malignancy and explored for their therapeutic potential in lymphoma, prostate, and bladder cancers. Additionally, gallium radioisotopes such as Ga-67 and Ga-68 are widely applied in diagnostic imaging, demonstrating gallium’s dual role in therapy and diagnostics (theranostics) (Darwesh et al., 2023).

Incorporation of Ga⁺³ into MOF architectures provides a unique opportunity to combine its therapeutic properties with the structural advantages of MOFs, including high loading capacity and controlled release behavior (Wang et al., 2025). Previous studies have shown that Ga-MOF systems can inhibit microbial growth and exhibit cytotoxic effects against cancer cells (Song et al., 2024). However, challenges remain regarding the use of biocompatible ligands, stability in physiological environments, and effective delivery platforms (Cordeiro Gomes et al., 2024).

To address these issues, we designed a novel Ga-MOF system using 4,4′,4″,4‴-(1,4-phenylenebis (pyridine-4,2,6-triyl))tetrabenzoic acid (H₄TBAPy) as the primary organic linker. This π-conjugated tetratopic ligand offers multiple coordination sites and enhanced framework stability (Cheng et al., 2024). In order to increase hydrophilicity and biological affinity, oxidized pectin, a naturally derived and biodegradable polysaccharide with aldehyde functionalities, was introduced as a secondary ligand. The resulting Ga-MOF/pectin complex was then embedded into a chitosan-based hydrogel matrix, providing additional biocompatibility, mucoadhesiveness, and inherent antimicrobial activity. Chitosan also enhances the material’s structural integrity and enables hydrogel formation suitable for biomedical environments.

The hybrid composite was synthesized via a microwave-assisted approach, offering a rapid, energy-efficient, and scalable route for material preparation (Li et al., 2024). Structural and physicochemical features were confirmed by SEM, BET, FT-IR, XRD, CHNO elemental analysis, and EDX mapping. Biological evaluations were performed to assess antibacterial activity against clinically relevant Gram-positive and Gram-negative bacteria and to investigate anticancer effects against A-431 (skin), MCF-7 (breast), and MG-63 (bone) cancer cell lines.

The distinctive novelty of this study lies in the first-time combination of H₄TBAPy and oxidized pectin as dual ligands in a Ga-based MOF, incorporated into a chitosan hydrogel via microwave-assisted synthesis, yielding a multifunctional biomaterial. This system synergistically integrates broad-spectrum antimicrobial effects, anticancer activity, biodegradability, and structural stability, highlighting its potential as a next-generation platform for applications in targeted therapy, wound healing, and oncology.

2 Materials and methods

2.1 Materials

Gallium nitrate (Ga(NO₃)₃·xH₂O, 99.9%) and the ligand 4,4′,4″,4‴-(1,4-phenylenebis (pyridine-4,2,6-triyl))tetrabenzoic acid (H₄TBAPy) were purchased from Sigma-Aldrich. Pectin (extracted from citrus peel) and chitosan (medium molecular weight, 75%–85% deacetylated) were obtained from Merck. All solvents used, including ethanol and distilled water, were of analytical grade and used without further purification. Microbial strains (Cryptococcus neoformans (ATCC 32045), Candida albicans (ATCC 10231), Fusarium oxysporum (ATCC 7601), Yersinia enterocolitica (ATCC 9610), Klebsiella pneumonia (ATCC 13883), Proteus mirabilis (ATCC 7002), Rhodococcus equi (ATCC 25729), Bacillus cereus (ATCC 11778), and Streptococcus agalactiae (ATCC 12386)) and cancer cell lines (skin (A-431), breast (MCF-7), and bone (MG-63)) were obtained from ATCC.

2.2 Synthesis of Ga-MOF

In a typical synthesis, 2 mmol of Ga(NO₃)₃·xH₂O and 1 mmol of H₄TBAPy were dissolved in 100 mL of a 1:1 ethanol-water mixture and stirred at 50 °C for 20 min. Then, 0.1 mmol of oxidized pectin was added dropwise and the solution was irradiated in a microwave reactor (Anton Paar Monowave, 300 W) for 20 min. The resulting precipitate was collected by centrifugation and washed thrice with ethanol–water (1:1 v/v). The solid was then dried under vacuum at room temperature for 24 h (ref).

2.3 Preparation of Ga-MOF/hydrogel

To prepare the Ga-MOF/Hydrogel, 100 mg of chitosan was dissolved in 20 mL of 1% acetic acid solution. Separately, 100 mg of the synthesized Ga-MOF composite was dispersed in 10 mL distilled water and ultrasonicated (Elma-S 30, 40 kHz, 100 W) for 15 min. The MOF dispersion was gradually added to the chitosan solution under magnetic stirring. After homogenization for 30 min, the mixture was transferred into molds and left to gel at 4 °C overnight. The hydrogel was subsequently neutralized using 0.1 M NaOH, washed with distilled water, and stored at 4 °C prior to characterization and biological assays (Al-Khafaji et al., 2023; Trombino et al., 2023).

2.4 Characterization

The chemical structure and functional groups were analyzed by FT-IR spectroscopy (Bruker Tensor II) in the 400–4,000 cm^-1 range. Crystalline structure was determined by powder X-ray diffraction (PXRD) using a PANalytical X’Pert PRO diffractometer with Cu Kα radiation (λ = 1.5406 Å). Surface morphology was observed by field emission scanning electron microscopy (FESEM, Tescan Mira3) coupled with energy-dispersive X-ray spectroscopy (EDX) and elemental mapping. Surface area and porosity were analyzed by Brunauer–Emmett–Teller (BET) method using a Micromeritics ASAP 2020 analyzer.

2.5 Antimicrobial assay

The antimicrobial activity of the Ga-MOF/Hydrogel was tested using both the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), and minimum bactericidal concentration (MBC) methods. The hydrogel samples were tested against three fungal species (C. neoformans/ATCC 32045, C. albicans/ATCC 10231, F. oxysporum/ATCC 7601), three Gram-negative bacterial strains (Y. enterocolitica/ATCC 9610, Klebsiella pneumoniae/ATCC 13883, P. mirabilis/ATCC 7002), and three Gram-positive bacterial strains (R. equi/ATCC 25729, B. cereus/ATCC 11778, S. agalactiae/ATCC 12386). Microbial suspensions were prepared at 10⁶ CFU/mL and exposed to various concentrations of the hydrogel in 96-well plates. After 24 h incubation at 37 °C, MIC was determined by visual turbidity, MFC and MBC was evaluated by plating 100 μL onto nutrient agar and observing colony formation (Mohammed Yaseen et al., 2025).

2.6 Anticancer assay

Cell viability was assessed using the MTT assay. A-431, MCF-7, and MG-63 cells were seeded in 96-well plates at a density of 1 × 10⁴ cells/well and incubated with various concentrations of the hydrogel composite for 24 and 48 h. Following treatment, MTT solution (5 mg/mL) was added and the plates were incubated for an additional 4 h. Formazan crystals were dissolved in DMSO and absorbance was read at 570 nm using a microplate reader (Moghaddam-Manesh and Hosseinzadegan, 2021; Muzammil et al., 2023).

3 Results and discussion

3.1 Synthesis and characterization

The Ga-MOF/Hydrogel was successfully synthesized via a microwave-assisted method using Ga(NO₃)₃ and H₄TBAPy as the primary building blocks, in combination with oxidized pectin as a secondary ligand (Figure 1). Since gallium has the ability to complex with three oxygens, and each H₄TBAPy has 4 O-H groups with ligand-donating properties, and another oxygen is provided by pectin, a metal/ligand ratio of 2/1 was chosen. The hybrid hydrogel was then obtained by integrating the resulting MOF into a chitosan matrix, yielding a composite with desirable physicochemical and biological properties, as confirmed by SEM analysis (Figure 2A), BET surface area measurement (Figure 2B), FT-IR spectra (Figure 2C), and XRD patterns (Figure 2D).

Figure 1

Figure 1. Ga-MOF/Hydrogel’s structure.

Figure 2

Figure 2. Ga-MOF/Hydrogel’s SEM (A), N₂ adsorption/desorption (B), FT-IR (C), XRD (D), EDAX (E), and EDAX mapping (F).

The microwave-assisted route allowed for rapid nucleation and crystallization of the MOF structure, while minimizing energy consumption and reaction time compared to conventional solvothermal synthesis (Phan et al., 2023; Zhao et al., 2023).

The presence of uniform MOF crystals within the hydrogel matrix was confirmed by SEM analysis (Figure 2A). SEM analysis revealed a porous morphology with aggregated crystalline domains embedded within the hydrogel matrix. The SEM images showed a uniform distribution of the MOF particles within the chitosan network, supporting the successful integration of the inorganic and polymeric components.

The BET surface area analysis revealed a specific surface area of 1,633 m²/g, reflecting a high degree of porosity and favorable adsorption capacity (Figure 2B). The total pore volume and average pore diameter were consistent with a predominantly microporous structure. Furthermore, the nitrogen adsorption/desorption isotherms displayed a type IV profile accompanied by a clear hysteresis loop, indicating the presence of mesoporous features (Baldovino-Medrano et al., 2023). The average pore diameter was calculated to be approximately 1.24 nm.

Elemental analysis (CHNO) confirmed the presence and appropriate distribution of carbon, hydrogen, nitrogen, and oxygen within the final structure, supporting the successful synthesis of the designed material supporting the successful synthesis of the designed material (Table 1). The analysis revealed that the sample contained 55.46% carbon, 5.73% hydrogen, 4.08% nitrogen, and 24.53% oxygen, indicating the effective incorporation of the expected organic and heteroatomic components into the framework.

Table 1

Table 1. Statistical analyses in anticancer activity of Ga-MOF/Hydrogel.

FT-IR analysis (Figure 2C) revealed characteristic vibrations indicative of metal-ligand coordination. The appearance of a peak around 3,300 cm^-1 indicated the presence of hydroxyl groups from chitosan. The C-H groups appeared in near 2,800 cm^-1. Peaks near 1,595 cm^-1 and 1,380 cm^-1 were attributed to carbonyl (C=O) and imine (C=N) groups, respectively. The peak around 1,100 cm^-1 indicated the presence of C-O groups. FT-IR spectra confirmed the coordination of Ga³⁺ ions with carboxylate (Ga-O) (Messeddek et al., 2025).

XRD analysis was employed to assess the crystallinity and phase purity of the synthesized Ga-MOF/Hydrogel (Figure 2D). The diffraction pattern exhibited sharp and intense peaks at 2θ values corresponding to the crystal planes (100), (110), (200), (211), (220) and (310), respectively. These reflections matched well with the JCPDS reference card No. 43–1,012, confirming the formation of a crystalline gallium-based coordination framework (Mi et al., 2012; Kumar et al., 2013). Crystallite size, calculated via the Scherrer equation (Hossain and Ahmed, 2023; Moghaddam-manesh et al., 2023). Based on this calculation, the average crystallite size was determined to be approximately 103 nm, indicating the nanocrystalline nature of the MOF structure.

EDAX (Figure 2E) and EDAX mapping (Figure 2F) further confirmed the presence and homogeneous distribution of Ga, C, N, and O elements throughout the composite. The high-resolution elemental maps supported the conclusion that both the metal nodes and the organic ligands were well distributed within the hydrogel.

These unique features, such as nanoscale crystallinity, high surface area, and porous structure, are closely tied to the synthetic strategy employed (Kumar et al., 2022). The use of microwave irradiation not only accelerated the reaction but also promoted uniform particle growth and avoided excessive agglomeration, which are common drawbacks in traditional solvothermal methods (Yin et al., 2023). This synthesis approach enabled better control over size and morphology, ultimately enhancing the adsorptive and biological functionalities of the final composite (Mu et al., 2024). Thus, the adopted synthesis method is not only energy-efficient but also crucial in engineering a MOF-based system with superior structural and functional characteristics (Jayaramulu et al., 2022).

3.2 Biological activity

The synthesized Ga-MOF/Hydrogel exhibit promising antimicrobial and anticancer potential, which can be attributed to their favorable physicochemical characteristics, such as a high specific surface area and the presence of biologically active constituents within their nanoscale structure. To evaluate these bioactivities, antimicrobial assays were conducted against fungal strains, as well as Gram-negative and Gram-positive bacteria. In parallel, cytotoxicity studies were performed on three distinct cancer cell lines. The findings and interpretations related to antimicrobial efficacy are presented in Section 3.2.1, while the results concerning anticancer activity are detailed in Section 3.2.2.

3.2.1 Antimicrobial activity

Figures 3A–C illustrate the antifungal and antibacterial activities of the Ga-MOF/Hydrogel composite against various Gram-negative and Gram-positive microbial strains.

Figure 3

Figure 3. Ga-MOF/Hydrogel’s antimicrobial activity: fungal (A), Gram-negative bacterial (B), and Gram-positive bacterial (C) strains.

The minimum inhibitory concentrations (MICs) for antifungal activity were determined to be 64 μg/mL against Cryptococcus neoformans (ATCC 32045), 8 μg/mL against Candida albicans (ATCC 10231), and 32 μg/mL against Fusarium oxysporum (ATCC 7601). For Gram-negative bacterial strains, MICs of 8, 16, and 8 μg/mL were observed against strains Yersinia enterocolitica (ATCC 9610), Klebsiella pneumonia (ATCC 13883), and Proteus mirabilis (ATCC 7002), respectively. In the case of Gram-positive bacteria, MIC values of 16, 32, and 8 μg/mL were recorded against strains Rhodococcus equi (ATCC 25729), Bacillus cereus (ATCC 11778), Streptococcus agalactiae (ATCC 12386), respectively.

To benchmark the antimicrobial efficacy of Ga-MOF/Hydrogel, its performance was compared with commonly used commercial drugs, nystatin, ketoconazole for antifungal evaluation and penicillin, ceftriaxone for antibacterial tests. Notably, ATCC 32045 showed resistance to nystatin, and ATCC 9610, ATCC 13883, ATCC 7002, and ATCC 11778 exhibited resistance to penicillin, and ATCC 11778 exhibited resistance to ceftriaxone. In contrast, Ga-MOF/Hydrogel demonstrated significant inhibitory effects against these resistant strains.

The superior antimicrobial performance of Ga-MOF/Hydrogel can be attributed to its high specific surface area, which enhances its interaction with microbial cells (Hamedi et al., 2024). Additionally, the synergistic presence of gallium, the organic linker, pectin, and chitosan, each known for their intrinsic antimicrobial properties, further contributes to its broad-spectrum activity (Freitas et al., 2021; Shi et al., 2023; Su et al., 2025).

These findings suggest that Ga-MOF/Hydrogel represents a multifunctional antimicrobial platform with potential applicability in combating resistant microbial infections.

3.2.2 Anticancer activity

To assess the anticancer efficacy of the synthesized Ga-MOF/Hydrogel, in vitro cytotoxicity studies were performed against skin (A-431), breast (MCF-7), and bone (MG-63) cancer cells. The results, illustrated in Figures 4A–C, respectively, demonstrate a dose- and time-dependent reduction in cell viability, highlighting the compound’s potent anticancer properties. The half-maximal inhibitory concentration (IC₅₀) values were determined to be 70.85 μg/mL for A-431, 52.24 μg/mL for MCF-7, and 54.47 μg/mL for MG-63 cancer cells at 24 h and 59.52 μg/mL, 44.48 μg/mL, and 39.46 μg/mL at 48 h, respectively.

Figure 4

Figure 4. Ga-MOF/Hydrogel’s anticancer activity: skin (A), breast (B), and bone (C) cancer cells.

The most pronounced cytotoxic effects were observed at the highest tested concentration (50 μg/mL) and the longest exposure duration (48 h), at which point the survival rates of A-431, MCF-7, and MG-63 cancer cells declined to 61%, 45%, and 41%, respectively.

Statistical analyses confirmed that both concentration and exposure time significantly influenced cell viability, as shown in Table 1, with corresponding p-values indicating strong statistical significance.

Analogous to its antimicrobial activity, the notable anticancer performance of the Ga-MOF/Hydrogel can be attributed to its nanoscale structure, offering a large specific surface area for cellular interaction, as well as the intrinsic bioactivity of its constituents, including gallium ions, polysaccharide-based ligands (e.g., chitosan and pectin), and the coordinated organic framework. These features likely enhance cellular uptake and disrupt cancer cell functions, contributing to the observed cytotoxicity (Li et al., 2022; Sultana, 2023; Yang et al., 2023; Zhang et al., 2023).

The observed bioactivity can be explained by the gradual release of Ga³⁺ ions from the hydrogel matrix, where swelling and partial degradation of chitosan/pectin allow a sustained diffusion–dissolution process; this mechanism is particularly relevant for topical applications, where moisture at the site of contact facilitates controlled release.

Based on the observed anticancer activity, it can be concluded that the synthesized Ga-MOF/Hydrogel nanocomposite not only exhibits substantial antimicrobial efficacy but also demonstrates significant cytotoxic effects against three distinct cancer cell lines. This multifunctional bioactivity highlights the unique therapeutic potential of the compound. Given its broad-spectrum antimicrobial action and pronounced anticancer properties, the synthesized material emerges as a promising candidate for the development of a novel therapeutic agent with multiple biological functions (Li et al., 2022; Sultana, 2023; Yang et al., 2023).

Although our Ga-MOF/hydrogel was benchmarked against conventional antibiotics and antifungal agents in their standard forms, future studies should include direct comparisons with conventional drug-based hydrogel formulations to better highlight the relative advantages of the MOF–gel matrix.

To further validate its applicability and safety, it is recommended that in vivo and in silico studies be conducted to elucidate its precise mechanisms of action, assess its cytotoxicity in normal tissues, and perform additional biochemical evaluations. Should these investigations confirm its biocompatibility and efficacy, the Ga-MOF/Hydrogel composite may be proposed as a novel multifunctional therapeutic platform for future biomedical applications.

4 Conclusion

In this study, a novel Ga-MOF/hydrogel composite was successfully synthesized through a microwave-assisted strategy, employing H₄TBAPy as a tetratopic ligand, oxidized pectin as a biopolymeric modifier, and chitosan as the hydrogel-forming matrix. Structural characterizations confirmed the formation of a porous and nanocrystalline coordination framework with high surface area, uniform morphology, and homogenous elemental distribution. The coordination between Ga³⁺ ions and carboxylate functionalities was clearly validated by FT-IR and XRD analyses, highlighting the structural integrity and crystalline nature of the resulting MOF embedded in a hydrogel matrix. Biological evaluations revealed that the hybrid composite exhibited potent broad-spectrum antimicrobial activity, including efficacy against drug-resistant microbial strains, as well as significant cytotoxic effects against skin, breast, and bone cancer cell lines. These findings underscore the synergistic contribution of gallium’s inherent bioactivity, the biocompatibility of pectin and chitosan, and the physicochemical advantages conferred by the MOF architecture. Despite its promising in vitro performance and cytotoxicity assessments on normal cells, certain limitations remain. In addition, detailed Ga 2p X-ray photoelectron spectroscopy (XPS) analysis was not included in the present work. Future experiments will incorporate Ga 2p_3/2 and Ga 2p_1/2 spectra to confirm the Ga³⁺ valence state and further substantiate its role in mimicking Fe³⁺ in multivalent systems. Notably, the long-term biocompatibility, biodegradation behavior, and pharmacokinetics of the composite have yet to be explored. Moreover, the exact mechanisms underlying the observed anticancer and antimicrobial effects require further elucidation through molecular-level investigations. The scalability of the synthesis method and its reproducibility across different batches also merit evaluation to ensure consistent material quality for clinical translation. A limitation of this study is that conventional drug-based hydrogel formulations were not included as direct controls; future investigations will address this gap to provide a clearer comparative evaluation. In light of these considerations, future research should focus on in vivo assessments, computational modeling of drug-host interactions, and controlled release profiling under physiological conditions. If validated through these extended studies, the Ga-MOF/hydrogel platform may serve as a versatile candidate for targeted therapeutic delivery, wound healing, or multifunctional biomedical implants.

Data availability statement

The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.

Author contributions

FtA: Formal Analysis, Writing – original draft, Writing – review and editing. FrF: Project administration, Writing – original draft, Writing – review and editing. FA: Data curation, Writing – original draft. NY: Supervision, Writing – review and editing. AS: Methodology, Writing – original draft. ZS: Investigation, Writing – review and editing. MA: Formal Analysis, Writing – original draft. MJ: Validation, Writing – review and editing. HM: Conceptualization, Writing – review and editing. AS: Resources, Writing – review and editing. KM: Validation, Writing – review and editing.

Funding

The author(s) declare that financial support was received for the research and/or publication of this article. The authors extend their appreciation to the Deanship of Research & Graduate Studies at King Khalid University, KSA, for funding this work through a research group program under grant number RGP. 2/659/46.

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Generative AI statement

The author(s) declare that no Generative AI was used in the creation of this manuscript.

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