Apr 23, 2026
Development of Carboxymethyl-Sagocellulose-Stabilized Fe3O4 Nanoparticles Conjugated with 5-Fluorouracil for Targeted Photothermal Therapy in Cancer Treatment
- Anand Kumar Veeramachineni1
- 1Springer Nature
Protocol Citation: Anand Kumar Veeramachineni 2026. Development of Carboxymethyl-Sagocellulose-Stabilized Fe3O4 Nanoparticles Conjugated with 5-Fluorouracil for Targeted Photothermal Therapy in Cancer Treatment. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ld67yng5b/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 18, 2025
Last Modified: April 23, 2026
Protocol Integer ID: 227583
Keywords: fluorouracil for targeted photothermal therapy, targeted photothermal therapy, stabilized fe3o4 nanoparticle, fe3o4 nanoparticle, photothermal therapy, tumor ablation, cancer treatment, development of carboxymethyl, carboxymethyl
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Abstract
To synthesize and evaluate carboxymethyl-sagocellulose-stabilized Fe3O4 nanoparticles conjugated with 5-fluorouracil (5-FU) for targeted photothermal therapy (PTT) in cancer treatment, assessing their efficacy in drug delivery and tumor ablation.
Materials
Materials -
Ethylene Glycol 40 mL
Sodium Acetate 3.0 g
Ferric Chloride 1.08 g
Poly(4-styrenesulfonic acid-co-maleic acid) 1.0 g
Sodium Hydroxide As required
Sodium Monochloroacetate As required
5-Fluorouracil (5-FU) As required
(3-Aminopropyl) Triethoxysilane (APS) 4 mL
EDC (1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide) As required
Sulfo-NHS (N-Hydroxysulfosuccinimide) As required
Phosphate Buffer Solution (PBS) As required
Deionized Water As required
Reagents -
Sodium Hydroxide
5-Fluorouracil (5-FU)
EDC
Sulfo-NHS
Safety warnings
Handle all chemicals with appropriate personal protective equipment (PPE). Dispose of waste according to institutional guidelines. Ensure proper ventilation when working with volatile solvents.
Synthesis of Fe3O4 Nanoparticles
Dissolve 1.08 g of ferric chloride hexahydrate and 3.0 g of sodium acetate in 40 mL of ethylene glycol.
Homogenize the solution and transfer it to an autoclave.
Heat at 200 °C for 10 hours.
Allow the solution to cool, then wash the precipitate with deionized water and dry at 30 °C overnight.
Surface Modification of Fe3O4 Nanoparticles
Disperse 0.60 g of Fe3O4 nanoparticles in 350 mL of ethanol and 3 mL of deionized water.
Add 4 mL of APS and stir for 12 hours under nitrogen atmosphere at room temperature.
Wash the precipitate with deionized water and dry at 30 °C overnight.
Conjugation of CMSC and 5-FU
Dissolve 1 g of CMSC in 200 mL of PBS (pH 7.0).
Add EDC and sulfo-NHS at a molar ratio of 0.5 relative to the carboxyl groups of CMSC.
Stir for 12 hours in the dark.
Dialyze against distilled water for 6 hours to remove unreacted EDC and sulfo-NHS.
Mix the activated CMSC with 5-FU and Fe3O4-NH2 nanoparticles, stir for 12 hours.
Recover the nanoparticles using a magnet, wash with deionized water, and dry.
Evaluation of Photothermal Effect
Incubate HeLa cells with Fe3O4-CMSC-5FU nanoparticles at concentrations of 75, 150, and 300 μg/mL for 12 hours.
Irradiate with an 808 nm laser at a power density of 2 W/cm² for 1-3 minutes.
Assess cell viability using MTT assay.