{
  "abstract": "Enzyme aggregation and polymer encapsulation are complementary strategies used to overcome the intrinsic instability and limited bioavailability of hydrophilic enzymes in medical uses. In this work, we report a systematic study of polymer-encapsulated cross-linked enzyme aggregates (pe-CLEAs) as a hierarchical nanostructured platform for the stabilization and delivery of galactosylceramidase (GALC), a lysosomal enzyme used in the enzyme replacement therapy for Krabbe disease. GALC-CLEAs were produced via cross-linking and controlled precipitation, yielding stable aggregates with small hydrodynamic diameters (40-66 nm) and narrow polydispersity index (PDI < 0.1). Scaling up the synthesis by 50-fold markedly enhanced total enzymatic output, reaching 1225.4 ± 213.2 U·mL⁻¹. GALC-CLEAs were subsequently encapsulated within PLGA nanoparticles. Systematic screening of enzyme-to-polymer ratios revealed an optimal formulation at 75 μg GALC per mg PLGA, displaying a maximum specific activity of 6.0 ± 1.3 U·mg⁻¹. Colloidal stability and retention assays showed that pe-CLEAs preserved full catalytic activity and retained enzyme confinement over 30 days at 4°C. At 37°C, pe-CLEAs exhibited time-dependent size dynamics while retaining substantial activity. In vitro evaluations in primary fibroblasts derived from Twitcher mice (a Krabbe disease model) demonstrated complete cytocompatibility up to 5.0 mg·mL⁻¹ and robust intracellular recovery of GALC activity across tested doses (0.5-5.0 mg·mL⁻¹), outperforming equivalent doses of native enzyme. This work provides a comprehensive physicochemical and functional characterization of pe-CLEAs, elucidating how hierarchical aggregation and polymer encapsulation can synergistically preserve stability and bioactivity. The design principles outlined here support the rational development of polymer-enzyme nanocarriers for enzyme replacement therapy and related biomedical applications.",
  "abstract_shown": true,
  "affiliations": [
    "Istituto Nanoscienze - CNR, NEST-SNS Piazza San Silvestro 12 56126 Pisa Italy.",
    "Interdisciplinary Research Center \"Health Science\" Scuola Superiore Sant'Anna Piazza Martiri Della Libertà, 33 56127 Pisa Italy sssup.it."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Laura Colagiorgio",
      "orcid": "0000-0002-4000-3973"
    },
    {
      "affs": [
        0
      ],
      "name": "Mariacristina Gagliardi",
      "orcid": "0000-0002-5860-3141"
    },
    {
      "affs": [
        0
      ],
      "name": "Ambra Del Grosso",
      "orcid": "0000-0002-1838-9064"
    },
    {
      "affs": [
        0
      ],
      "name": "Sara Carpi",
      "orcid": "0000-0002-3291-9009"
    },
    {
      "affs": [
        0
      ],
      "name": "Miriam De Sarlo"
    },
    {
      "affs": [
        0
      ],
      "name": "Marco Cecchini",
      "orcid": "0000-0002-9688-2356"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4ecv7bzkqdg5e.bib",
  "cited_by": 0,
  "doi": "10.1155/ijbm/8346433",
  "fulltext_url": "https://europepmc.org/articles/PMC13678281",
  "id": "4ecv7bzkqdg5e",
  "issns": [
    "1687-8787",
    "1687-8795"
  ],
  "landing_url": "https://europepmc.org/article/MED/42898837",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7222944266",
  "pdf_url": "https://europepmc.org/articles/PMC13678281?pdf=render",
  "pmcid": "PMC13678281",
  "pmid": "42898837",
  "published": "2026-10-18",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/4ecv7bzkqdg5e.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:19:33Z",
      "native_id": "MED:42898837",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42898837"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "PLGA-Encapsulated Cross-Linked GALC Aggregates as a Hierarchical Nanostructured Platform for Enzyme Stabilization and Delivery.",
  "title_text": "PLGA-Encapsulated Cross-Linked GALC Aggregates as a Hierarchical Nanostructured Platform for Enzyme Stabilization and Delivery.",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:19:33Z",
  "url": "https://research.draconitic.com/paper/4ecv7bzkqdg5e",
  "venue": "International journal of biomaterials",
  "versions": []
}