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Cited by 114Electrospun Nanofibers in Biomedical ApplicationsTissue Engineering and Regenerative MedicineSilk-based biomaterials and applications

Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro

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Brown, Jessica H., Das, Prativa, DiVito, Michael D., Ivancic, David, Tan, Lay Poh, Wertheim, Jason A.·2018·Acta Biomaterialia
DOI 10.1016/j.actbio.2018.02.009PMC PMC5985221
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Abstract

A major challenge of maintaining primary hepatocytes in vitro is progressive loss of hepatocyte-specific functions, such as protein synthesis and cytochrome P450 (CYP450) catalytic activity. We developed a three-dimensional (3D) nanofibrous scaffold made from poly(l-lactide-co-glycolide) (PLGA) polymer using a newly optimized wet electrospinning technique that resulted in a highly porous structure that accommodated inclusion of primary human hepatocytes. Extracellular matrix (ECM) proteins (type I collagen or fibronectin) at varying concentrations were chemically linked to electrospun PLGA using amine coupling to develop an in vitro culture system containing the minimal essential ECM components of the liver micro-environment that preserve hepatocyte function in vitro. Cell-laden nanofiber scaffolds were tested in vitro to maintain hepatocyte function over a two-week period. Incorporation of type I collagen onto PLGA scaffolds (PLGA-C high: 100 µg/mL) led to 10-fold greater albumin secretion, 4-fold higher urea synthesis, and elevated transcription of hepatocyte-specific CYP450 genes (CYP3A4, 3.5-fold increase and CYP2C9, 3-fold increase) in primary human hepatocytes compared to the same cells grown within unmodified PLGA scaffolds over two weeks. These indices, measured using collagen-bonded scaffolds, were also higher than scaffolds coupled to fibronectin or an ECM control sandwich culture composed of type I collagen and Matrigel. Induction of CYP2C9 activity was also higher in these same type I collagen PLGA scaffolds compared to other ECM-modified or unmodified PLGA constructs and was equivalent to the ECM control at 7 days. Together, we demonstrate a minimalist ECM-based 3D synthetic scaffold that accommodates primary human hepatocyte inclusion into the matrix, maintains long-term in vitro survival and stimulates function, which can be attributed to coupling of type I collagen. Statement of significance Culturing primary hepatocytes within a three-dimensional (3D) structure that mimics the natural liver environment is a promising strategy for extending the function and viability of hepatocytes in vitro. In the present study we generate porous PLGA nanofibers, that are chemically modified with extracellular matrix proteins, to serve as 3D scaffolds for the in vitro culture of primary human hepatocytes. Our findings demonstrate that the use of ECM proteins, especially type I collagen, in a porous 3D environment helps to improve the synthetic function of primary hepatocytes over time. We believe the work presented within will provide insights to readers for drug toxicity and tissue engineering applications.

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  • Electrospun scaffolds with pore sizes and structures similar to native liver tissue often outperform non-porous or unmodified materials, permitting additional albumin secretion, urea synthesis, and CYP450 activity.

    “Electrospun scaffolds with pore sizes and structures similar to native liver tissue often outperform non-porous or unmodified materials, permitting additional albumin secretion, urea synthesis, and CYP450 activity [28].”

    From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [28]

    Results §3.1 reports wet-electrospun pores approximating those of acellular human liver tissue. Results §§3.5–3.6 report higher albumin secretion, urea synthesis, and CYP450 activity in collagen-modified scaffolds than in unmodified PLGA scaffolds. The paper does not establish that liver-like pore structure itself caused all three improvements, compare these outcomes with non-porous materials, or support the broader claim that such scaffolds often outperform other materials. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • Porous PLGA scaffolds modified with a coating of ECM components such as collagen greatly enhance hepatocyte adhesion and function.

    “Porous polymeric scaffolds composed of poly (lactic-co-glycolic acid) (PLGA) and modified with a coating of ECM components such as collagen have greatly enhanced hepatocyte adhesion and function [28].”

    From The Hepatocyte Expansion Paradox: A Review of In Vitro Challenges and Advances (2026) · cites this paper as [28]

    The Abstract reports substantially higher albumin secretion, urea synthesis, and hepatocyte-specific gene expression in primary human hepatocytes cultured in collagen-modified PLGA scaffolds than in unmodified scaffolds. The Introduction says the ECM modification was intended to provide adhesive support, and the Results report more cell–cell contacts in ECM-modified scaffolds, but our copy does not establish a greatly enhanced rate of hepatocyte adhesion to the scaffold. (LLM, conf=0.84)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 39

  1. [1]Hu C Li L In vitro culture of isolated primary hepatocytes and stem cell-derived hepatocyte-like cells for liver regeneration Protein Cell 6 8 2015 562 574 26088193 PMC4506286
  2. [2]Rowe C Gerrard DT Jenkins R Berry A Durkin K Sundstrom L Goldring CE Park BK Kitteringham NR Hanley KP Hanley NA Proteome-wide analyses of human hepatocytes during differentiation and dedifferentiation Hepatology 58 2 2013 799 809 23526496 PMC3842115
  3. [4]Dunn JC Yarmush ML Koebe HG Tompkins RG Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich configuration FASEB J 3 2 1989 174 7 2914628
  4. [7]Berthiaume F Moghe PV Toner M Yarmush ML Effect of extracellular matrix topology on cell structure, function, and physiological responsiveness: hepatocytes cultured in a sandwich configuration FASEB J 10 13 1996 1471 84 8940293
  5. [8]Wang X Ding B Li B Biomimetic electrospun nanofibrous structures for tissue engineering Mater Today 16 6 2013 229 241 PMC4130655 25125992
  6. [9]Lim SH Mao HQ Electrospun scaffolds for stem cell engineering Adv Drug Delivery Rev 61 12 2009 1084 1096 19647024
  7. [10]Bhardwaj N Kundu SC Electrospinning: A fascinating fiber fabrication technique Biotechnol Adv 28 3 2010 325 347 20100560
  8. [11]Kitsara M Agbulut O Kontziampasis D Chen Y Menasché P Fibers for hearts: A critical review on electrospinning for cardiac tissue engineering Acta Biomater 48 2017 20 40 27826001
  9. [12]Chen N Tian L He L Ramakrishna S Nanobiomaterials for neural regeneration Neural Regener Res 11 9 2016 1372 1374 PMC5090823 27857724
  10. [13]Rezvani Z Venugopal JR Urbanska AM Mills DK Ramakrishna S Mozafari M A bird’s eye view on the use of electrospun nanofibrous scaffolds for bone tissue engineering: Current state- of- the- art, emerging directions and future trends Nanomedicine 12 7 2016 2181 2200 27247186
  11. [15]Khorshidi S Solouk A Mirzadeh H Mazinani S Lagaron JM Sharifi S Ramakrishna S A review of key challenges of electrospun scaffolds for tissue-engineering applications Journal of tissue engineering and regenerative medicine 10 9 2016 715 38 25619820
  12. [18]Hiraoka Y Kimura Y Ueda H Tabata Y Fabrication and biocompatibility of collagen sponge reinforced with poly(glycolic acid) fiber Tissue Eng 9 6 2003 1101 12 14670098
  13. [19]Baiocchini A Montaldo C Conigliaro A Grimaldi A Correani V Mura F Ciccosanti F Rotiroti N Brenna A Montalbano M D’Offizi G Capobianchi MR Alessandro R Piacentini M Schinina ME Maras B Del Nonno F Tripodi M Mancone C Extracellular Matrix Molecular Remodeling in Human Liver Fibrosis Evolution PloS one 11 3 2016 PMC4801190 26998606
  14. [20]Azuma H Paulk N Ranade A Dorrell C Al-Dhalimy M Ellis E Strom S Kay MA Finegold M Grompe M Robust expansion of human hepatocytes in Fah(−/−)/Rag2(−/−)/Il2rg (−/−) mice Nat Biotechnol 25 8 2007 903 910 17664939 PMC3404624
  15. [21]Uzarski JS DiVito MD Wertheim JA Miller WM Essential design considerations for the resazurin reduction assay to noninvasively quantify cell expansion within perfused extracellular matrix scaffolds Biomaterials 129 2017 163 175 28343003 PMC5765551
  16. [22]MacArthur R Leister W Veith H Shinn P Southall N Austin CP Inglese J Auld DS Monitoring Compound Integrity With Cytochrome P450 Assays and qHTS Journal of Biomolecular Screening 14 5 2009 538 546 19483146 PMC3430136
  17. [23]Meisenheimer PL Uyeda HT Ma D Sobol M McDougall MG Corona C Simpson D Klaubert DH Cali JJ Proluciferin Acetals as Bioluminogenic Substrates for Cytochrome P450 Activity and Probes for CYP3A Inhibition Drug Metabolism and Disposition 39 12 2011 2403 2410 21890735
  18. [24]Higuchi A Tsukamoto Y Cell separation of hepatocytes and fibroblasts through surface-modified polyurethane membranes Journal of biomedical materials research Part A 71 3 2004 470 9 15386485
  19. [25]Uygun BE Soto-Gutierrez A Yagi H Izamis ML Guzzardi MA Shulman C Milwid J Kobayashi N Tilles A Berthiaume F Hertl M Nahmias Y Yarmush ML Uygun K Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix Nature medicine 16 7 2010 814 20 PMC2930603 20543851
  20. [26]Chen HZ Peng Y Wu SC Tan LP Electrospun 3D Fibrous Scaffolds for Chronic Wound Repair Materials 9 4 2016 PMC5502965 28773394
  21. [27]Arterburn LM Zurlo J Yager JD Overton RM Heifetz AH A morphological study of differentiated hepatocytes in vitro Hepatology 22 1 1995 175 187 7601410
  22. [28]Olsavsky Goyak KM Laurenzana EM Omiecinski CJ Hepatocyte differentiation Methods in molecular biology (Clifton, NJ) 640 2010 115 38 PMC6690360 20645049
  23. [29]Smith HS Opioid Metabolism Mayo Clin Proc 84 7 2009 613 624 19567715 PMC2704133
  24. [30]Vorrink SU Ullah S Schmidt S Nandania J Velagapudi V Beck O Ingelman-Sundberg M Lauschke VM Endogenous and xenobiotic metabolic stability of primary human hepatocytes in long-term 3D spheroid cultures revealed by a combination of targeted and untargeted metabolomics FASEB journal: official publication of the Federation of American Societies for Experimental Biology 31 6 2017 2696 2708 28264975 PMC5434660
  25. [31]Madan A Graham RA Carroll KM Mudra DR Burton LA Krueger LA Downey AD Czerwinski M Forster J Ribadeneira MD Gan LS LeCluyse EL Zech K Robertson P Koch P Antonian L Wagner G Yu L Parkinson A Effects of Prototypical Microsomal Enzyme Inducers on Cytochrome P450 Expression in Cultured Human Hepatocytes Drug Metabolism and Disposition 31 4 2003 421 431 12642468
  26. [32]Haque A Gheibi P Gao Y Foster E Son KJ You J Stybayeva G Patel D Revzin A Cell biology is different in small volumes: endogenous signals shape phenotype of primary hepatocytes cultured in microfluidic channels 6 2016 33980 PMC5041105 27681582
  27. [33]Rajendran D Hussain A Yip D Parekh A Shrirao A Cho CH Long-Term Liver-Specific Functions of Hepatocytes in Electrospun Chitosan Nanofiber Scaffolds Coated with Fibronectin J Biomed Mater Res, Part A 105A 2017 2119 2128 28371246
  28. [34]Kim MH Kumar SK Shirahama H Seo J Lee JH Cho NJ Phenotypic regulation of liver cells in a biofunctionalized three-dimensional hydrogel platform Integr Biol 8 2 2016 156 166 26792030
  29. [35]Soars MG McGinnity DF Grime K Riley RJ The pivotal role of hepatocytes in drug discovery Chem-Biol Interact 168 1 2007 2 15 17208208
  30. [36]Li J Li L Yu H Cao H Gao C Gong Y Growth and metabolism of human hepatocytes on biomodified collagen poly(lactic-co-glycolic acid) three-dimensional scaffold ASAIO J 52 3 2006 321 7 16760723
  31. [37]Török E Lutgehetmann M Bierwolf J Melbeck S Düllmann J Nashan B Ma PX Pollok JM Primary human hepatocytes on biodegradable poly(l-lactic acid) matrices: A promising model for improving transplantation efficiency with tissue engineering Liver Transplantation 17 2 2011 104 114 21280182
  32. [38]Martini WZ Wolf SE Chinkes DL Chung KK Dubick MA Blackbourne L Yu YM Enhanced albumin synthesis in severely burned adults Shock 34 4 2010 364 8 20160662
  33. [39]Sohlenius-Sternbeck AK Determination of the hepatocellularity number for human, dog, rabbit, rat and mouse livers from protein concentration measurements Toxicol In Vitro 20 8 2006 1582 6 16930941
  34. [40]Sawada N Tomomura A Sattler CA Sattler GL Kleinman HK Pitot HC EFFECTS OF EXTRACELLULAR-MATRIX COMPONENTS ON THE GROWTH AND DIFFERENTIATION OF CULTURED RAT HEPATOCYTES In Vitro Cell Dev Biol 23 4 1987 267 273 2883170
  35. [41]Sawada N Tomomura A Sattler CA Sattler GL Kleinman HK Pitot HC EXTRACELLULAR-MATRIX COMPONENTS INFLUENCE DNA-SYNTHESIS OF RAT HEPATOCYTES IN PRIMARY CULTURE Exp Cell Res 167 2 1986 458 470 3490392
  36. [42]Sanchez A Alvarez AM Pagan R Roncero C Vilaro S Benito M Fabregat I Fibronectin regulates morphology, cell organization and gene expression of rat fetal hepatocytes in primary culture J Hepatol 32 2 2000 242 250 10707864
  37. [43]Song W Lu Y-C Frankel AS An D Schwartz RE Ma M Engraftment of human induced pluripotent stem cell-derived hepatocytes in immunocompetent mice via 3D co-aggregation and encapsulation Sci Rep 5 2015 16884 26592180 PMC4655358
  38. [44]Wang Y Kim MH Shirahama H Lee JH Ng SS Glenn JS Cho NJ ECM proteins in a microporous scaffold influence hepatocyte morphology, function, and gene expression Sci Rep 6 2016 PMC5126637 27897167
  39. [45]Wang Y Lee JH Shirahama H Seo J Glenn JS Cho NJ Extracellular Matrix Functionalization and Huh-7.5 Cell Coculture Promote the Hepatic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells in a 3D ICC Hydrogel Scaffold ACS Biomater Sci Eng 2 12 2016 2255 2265 33465898

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Authors
Brown, J. H., Das, P., DiVito, M. D., Ivancic, D., Tan, L. P., & Wertheim, J. A.
Affiliations
  • Northwestern University (US)
  • Nanyang Technological University (SG)
Corresponding author
  • Tan, Lay Poh
  • Wertheim, Jason A.
ORCID
Venue
Acta Biomaterialia
Publisher
Elsevier BV
Published
2018-06
Volume
73
Pages
217-227
ISSN
1742-7061 (ISSN-L)
DOI
10.1016/j.actbio.2018.02.009
PMC
PMC5985221
OpenAlex
W2791649331
Cited by
114
References
2
Citations by year
2026: 3 · 2025: 10 · 2024: 12 · 2023: 11 · 2022: 17 · 2021: 15 · 2020: 22 · 2019: 17 · 2018: 7
FWCI
4.94
Type
journal article
Language
en
Open access
Licence
Locations
  • Acta Biomaterialia (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • DR-NTU (Nanyang Technological University) (submittedVersion)
Funders
  • Northwestern University
  • National Institutes of Health (TL1TR001423)
  • National Cancer Institute (P30CA060553)
  • National Institute of Diabetes and Digestive and Kidney Diseases (K08DK101757, T32DK077662)
  • National Center for Advancing Translational Sciences
Topics
  • Physical Sciences › Materials Science › Biomaterials › Electrospun Nanofibers in Biomedical Applications
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Physical Sciences › Materials Science › Biomaterials › Silk-based biomaterials and applications
Keywords
PLGA nanofibers, electrospun scaffolds, type I collagen, primary human hepatocytes, hepatocyte function, extracellular matrix, fibronectin, cytochrome P450, CYP3A4, CYP2C9, albumin secretion, urea synthesis, 3D cell culture, cell-laden scaffolds, hepatocyte viability
MeSH terms
Polylactic Acid-Polyglycolic Acid Copolymer, Animals, Cell Survival, Extracellular Matrix, Humans, Mice, Knockout, Hepatocytes, Collagen Type I, Mice, Tissue Scaffolds

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