Chitosan is a cationic polysaccharide derived from chitin, the second most abundant natural polymer on Earth after cellulose and the structural backbone of crustacean exoskeletons. Chemically, chitosan consists of repeating units of D-glucosamine and N-acetyl-D-glucosamine linked by beta-(1,4) glycosidic bonds, and its native amine groups become protonated at physiological pH. This positive charge allows the polymer to interact electrostatically with negatively charged cell membranes, modulating cell adhesion, migration and proliferation. Because of these properties, chitosan has long attracted interest as a wound-dressing material that can promote tissue regeneration, stimulate fibroblast activity, accelerate hemostasis and suppress microbial infection. The single most important quality parameter is the degree of deacetylation, the proportion of deacetylated glucosamine units in the chain; a chitin sample with more than 50 percent deacetylation is generally classified as chitosan, and higher values typically improve solubility, antimicrobial performance and biological interactions.

The catch has always been how to make it. Conventional extraction subjects shrimp shell powder to concentrated sodium hydroxide at high temperatures for hours, an energy-intensive process that risks degrading the polymer backbone and inflating production costs. Alternative technologies each carry their own burdens: ultrasound can accelerate extraction but may cleave polymer chains and reduce molecular weight, enzyme-assisted routes offer selectivity at the price of costly catalysts and long processing times, and deep eutectic solvents reduce harsh chemical use but complicate solvent recovery at industrial scale. Microwave-assisted extraction promises shorter reaction times, lower electricity consumption, simpler equipment and easier scalability, because microwave radiation heats the reaction mixture directly and uniformly through dipole rotation and ionic conduction rather than through slow conduction and convection.

The research team, working with shrimp shell waste from Indonesian processing of Litopenaeus vannamei, ground and sieved the dried shells to 74 to 150 micrometers and then optimized each of the three extraction stages under microwave irradiation. Demineralization with hydrochloric acid proved to be governed mainly by acid concentration rather than microwave power: at 0.5 M HCl more than 72 percent of the calcium remained, while both 1 M and 2 M HCl achieved complete calcium removal across 400, 500 and 600 watts within just five minutes. Deproteinization with sodium hydroxide followed a similar pattern, with protein removal rising from 99.18 percent at 2 percent NaOH to a full 100 percent at 10 percent NaOH, regardless of the power level applied.

Deacetylation, the step that transforms chitin into chitosan, responded strongly to both alkali concentration and microwave power. At a fixed 600 watts, increasing NaOH from 40 to 80 weight percent pushed the degree of deacetylation toward 80 percent even in five-minute treatments, and extending the reaction to ten minutes at 600 watts and 80 percent NaOH lifted the value to nearly 85 percent. FTIR spectroscopy, using the ratio of absorbance bands at 1320 and 1420 inverse centimeters, confirmed a degree of deacetylation of 85 percent for microwave-synthesized chitosan versus 80 percent for the conventionally produced material. More striking was the yield: microwave processing converted 10 percent of the starting shell mass into final chitosan, compared with only 1.6 percent for the conventional route, a six-fold improvement in material efficiency.

Structural characterization reinforced the case for the microwave route. X-ray diffraction showed that both products were semi-crystalline, but the conventionally synthesized sample exhibited a broadened and weakened peak near 2 theta of 10 degrees, indicating loss of crystallinity, whereas the microwave product retained a well-defined crystalline peak, suggesting that rapid volumetric heating minimizes disruption of the polymer backbone. Scanning electron microscopy revealed characteristic lamellar flake morphologies with smooth surfaces and dense packing for both materials, consistent with a high density of free amino groups. At the molecular level, the researchers explain the difference through kinetics: hydroxide ions attack the electrophilic carbonyl carbon of chitin’s acetamide groups, cleaving the C–N bond and releasing acetate, and microwave-driven dipolar heating accelerates this nucleophilic attack throughout the crystalline core instead of forcing slow diffusion from a hot surface inward.

The biological evaluation focused on RAW 264.7 murine macrophage-like cells, key players in the inflammatory phase of wound repair. In MTT cytotoxicity assays, microwave-derived chitosan at concentrations from 0.1 to 10 milligrams per milliliter did not significantly reduce cell viability over 24 or 48 hours of incubation, demonstrating excellent cytocompatibility. In scratch assays, monolayers cultured on the chitosan-coated substrates showed progressive wound closure beginning around 12 hours, with the cell-free gap nearly completely closed after 48 hours. The authors attribute this activity in part to positively charged calcium ions released from the biopolymer, which may enhance cell proliferation, and to the hydrophilic character of the chitosan surface. They caution, however, that macrophage results represent only one facet of a process that also depends on fibroblasts, keratinocytes, endothelial cells and extracellular matrix remodeling, and that fibroblast and keratinocyte models plus in vivo studies are needed to confirm therapeutic efficacy.

To probe mechanism, the team performed molecular docking of chitosan malate against AKT1, a protein kinase central to the PI3K/AKT signaling pathway that drives cell proliferation, migration and angiogenesis during tissue repair. Chitosan malate showed a predicted binding affinity of minus 9.6 kilocalories per mole toward AKT1, stronger than the known AKT activator SC79 at minus 7.9, with predicted contacts at residues Thr81, Thr82 and Arg273 in the PH domain and near the kinase domain interface. The researchers stress that these are exploratory computational estimates: the study did not measure AKT1 expression or phosphorylation, so any involvement of the pathway in the observed wound-closure behavior remains a hypothesis requiring experimental validation.

The economic case may prove equally consequential. By integrating equipment power ratings over operating times, the team calculated that microwave-assisted synthesis consumed up to 94.6 percent less energy than conventional heating. A techno-economic analysis anchored to Indonesian market conditions yielded a positive net present value of approximately USD 10.73 million, an annual profitability investment of 18.62 percent, and a benefit-cost ratio of 17.72, comfortably above the feasibility threshold of unity. Sensitivity analyses varying the product selling price by plus or minus 15 percent showed that the project remained profitable even in the worst scenario, with NPV near USD 9 million and a benefit-cost ratio of 15. Taken together, the results position microwave extraction as a commercially viable, energy-efficient pathway for turning one of the seafood industry’s largest waste streams into a biomedical raw material, though the authors note that pilot-scale validation, molecular-weight characterization, in vivo testing and continuous reactor design remain the next hurdles before clinical and industrial adoption.

Subject of Research: Microwave-assisted synthesis of high-quality chitosan from shrimp shell biowaste for wound-healing applications

Article Title: Comparative study of microwave-assisted and conventional methods for the synthesis of high-quality chitosan from shrimp shell biowaste to manipulate cell growth and functions: Supported molecular docking analysis

Article References: Rahmatunnisa, C., Budi, C. S., Ilhami, F. B., Puspitarini, S., Melanie, M., Hapidin, D. A., Kuo, D.-H., Zeleke, M. A., Kurnia, I., Hastuti, L. P., Rahmah, D. M., & Gultom, N. S. (2026). Comparative study of microwave-assisted and conventional methods for the synthesis of high-quality chitosan from shrimp shell biowaste to manipulate cell growth and functions: Supported molecular docking analysis. Case Studies in Chemical and Environmental Engineering, 14, Article 101481. https://doi.org/10.1016/j.cscee.2026.101481

Image Credits: AI Generated

DOI: 10.1016/j.cscee.2026.101481

Keywords: chitosan, shrimp shell waste, microwave-assisted extraction, chitin deacetylation, wound healing, biomaterials, molecular docking, AKT1, techno-economic analysis, biocompatibility, energy efficiency, marine biowaste