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A proteomic analysis of the interactions between poly(L-lactic acid) nanofibers and SH-SY5Y neuronal-like cells

  • Received: 09 September 2016 Accepted: 06 November 2016 Published: 14 November 2016
  • Poly (L-lactic acid) (PLLA) is a biodegradable and biocompatible polymer that has been put forward as a promising material for therapeutic approaches aiming to restore neuronal function. The topographic cues present in PLLA-based scaffolds, defined by the technique used in their preparation, have been shown to play a role on the cellular behavior of adherent cells. Even though this interaction has been shown to influence the regenerative output of the scaffold, there is a lack of studies addressing this response at the proteomic level. Hence, this work focuses on the effect of electrospun PLLA-based nanofibers on the proteome, cellular processes and signaling pathways of SH-SY5Y neuroblastoma cells. It also further explores how these molecular mediators might influence cell proliferation and differentiation upon in vitro culture. For that, mass spectrometry followed by bioinformatics analysis was firstly performed and further complemented with Western blot, cell viability and imaging assays. Results show that PLLA nanofibers differentially activate and inhibit specific cellular functions and signaling pathways related to cell division, apoptosis, actin remodeling, among others. These ultimately block cellular proliferation and induce morphological rearrangements through cytoskeleton remodeling, adaptations that turn cells more prone to differentiate. In synthesis, PLLA nanofibers shift the SH-SY5Y cells proteome towards a state more responsive to differentiation-inductive cues such as the retinoic acid. Unveiling cells responses to nanomaterials is an important step to increase the tools available for their manipulation and potentiate their use in neural tissue engineering. Further studies should be performed to compare the effects of other topographic cues on cellular behavior.

    Citation: Ana Marote, Nathalie Barroca, Rui Vitorino, Raquel M. Silva, Maria H.V. Fernandes, Paula M. Vilarinho, Odete A.B. da Cruz e Silva, Sandra I. Vieira. A proteomic analysis of the interactions between poly(L-lactic acid) nanofibers and SH-SY5Y neuronal-like cells[J]. AIMS Molecular Science, 2016, 3(4): 661-682. doi: 10.3934/molsci.2016.4.661

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  • Poly (L-lactic acid) (PLLA) is a biodegradable and biocompatible polymer that has been put forward as a promising material for therapeutic approaches aiming to restore neuronal function. The topographic cues present in PLLA-based scaffolds, defined by the technique used in their preparation, have been shown to play a role on the cellular behavior of adherent cells. Even though this interaction has been shown to influence the regenerative output of the scaffold, there is a lack of studies addressing this response at the proteomic level. Hence, this work focuses on the effect of electrospun PLLA-based nanofibers on the proteome, cellular processes and signaling pathways of SH-SY5Y neuroblastoma cells. It also further explores how these molecular mediators might influence cell proliferation and differentiation upon in vitro culture. For that, mass spectrometry followed by bioinformatics analysis was firstly performed and further complemented with Western blot, cell viability and imaging assays. Results show that PLLA nanofibers differentially activate and inhibit specific cellular functions and signaling pathways related to cell division, apoptosis, actin remodeling, among others. These ultimately block cellular proliferation and induce morphological rearrangements through cytoskeleton remodeling, adaptations that turn cells more prone to differentiate. In synthesis, PLLA nanofibers shift the SH-SY5Y cells proteome towards a state more responsive to differentiation-inductive cues such as the retinoic acid. Unveiling cells responses to nanomaterials is an important step to increase the tools available for their manipulation and potentiate their use in neural tissue engineering. Further studies should be performed to compare the effects of other topographic cues on cellular behavior.


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    [1] Seil JT, Webster TJ (2010) Electrically active nanomaterials as improved neural tissue regeneration scaffolds. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2: 635-647. doi: 10.1002/wnan.109
    [2] Quan Q, Chang B, Meng HY, et al. (2016) Use of electrospinning to construct biomaterials for peripheral nerve regeneration. Rev Neurosci.27: 761-768
    [3] Tian L, Prabhakaran MP, Ramakrishna S (2015) Strategies for regeneration of components of nervous system: scaffolds, cells and biomolecules. Regen Biomater 2: 31-45. doi: 10.1093/rb/rbu017
    [4] Yang F, Murugan R, Wang S, et al. (2005) Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neural tissue engineering. Biomaterials 26: 2603-2610. doi: 10.1016/j.biomaterials.2004.06.051
    [5] Wang HB, Mullins ME, Cregg JM, et al. (2009) Creation of highly aligned electrospun poly-L-lactic acid fibers for nerve regeneration applications. J Neural Eng 6: 016001. doi: 10.1088/1741-2560/6/1/016001
    [6] Morelli S, Salerno S, Piscioneri A, et al. (2010) Influence of micro-patterned PLLA membranes on outgrowth and orientation of hippocampal neurites. Biomaterials 31: 7000-7011. doi: 10.1016/j.biomaterials.2010.05.079
    [7] Callahan LA, Xie S, Barker IA, et al. (2013) Directed differentiation and neurite extension of mouse embryonic stem cell on aligned poly(lactide) nanofibers functionalized with YIGSR peptide. Biomaterials 34: 9089-9095. doi: 10.1016/j.biomaterials.2013.08.028
    [8] Borgens RB (1999) Electrically mediated regeneration and guidance of adult mammalian spinal axons into polymeric channels. Neuroscience 91: 251-264. doi: 10.1016/S0306-4522(98)00584-3
    [9] Ding Y, Yan Q, Ruan JW, et al. (2009) Electro-acupuncture promotes survival, differentiation of the bone marrow mesenchymal stem cells as well as functional recovery in the spinal cord-transected rats. BMC Neurosci 10: 35. doi: 10.1186/1471-2202-10-35
    [10] Guo B, Finne-Wistrand A, Albertsson AC (2010) Molecular architecture of electroactive and biodegradable copolymers composed of polylactide and carboxyl-capped aniline trimer. Biomacromolecules 11: 855-863. doi: 10.1021/bm9011248
    [11] Yang F, Murugan R, Ramakrishna S, et al. (2004) Fabrication of nano-structured porous PLLA scaffold intended for nerve tissue engineering. Biomaterials 25: 1891-1900. doi: 10.1016/j.biomaterials.2003.08.062
    [12] Yang IH, Co CC, Ho CC (2011) Controlling neurite outgrowth with patterned substrates. J Biomed Mater Res A 97: 451-456.
    [13] Lee YS, Collins G, Arinzeh TL (2011) Neurite extension of primary neurons on electrospun piezoelectric scaffolds. Acta Biomater 7: 3877-3886. doi: 10.1016/j.actbio.2011.07.013
    [14] Lee YS, Arinzeh TL (2012) The influence of piezoelectric scaffolds on neural differentiation of human neural stem/progenitor cells. Tissue Eng Part A 18: 2063-2072. doi: 10.1089/ten.tea.2011.0540
    [15] He L, Liao S, Quan D, Ma K, et al. (2010) Synergistic effects of electrospun PLLA fiber dimension and pattern on neonatal mouse cerebellum C17.2 stem cells. Acta Biomaterialia 6: 2960-2969.
    [16] Fukada E (2000) History and recent progress in piezoelectric polymers. IEEE Trans Ultrason Ferroelectr Freq Control 47: 1277-1290. doi: 10.1109/58.883516
    [17] Corey JM, Gertz CC, Wang BS, et al. (2008) The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons. Acta Biomater 4: 863-8675. doi: 10.1016/j.actbio.2008.02.020
    [18] He L, Liao S, Quan D, et al. (2010) Synergistic effects of electrospun PLLA fiber dimension and pattern on neonatal mouse cerebellum C17.2 stem cells. Acta Biomater 6: 2960-2969. doi: 10.1016/j.actbio.2010.02.039
    [19] Evans GR, Brandt K, Widmer MS, et al. (1999) In vivo evaluation of poly(L-lactic acid) porous conduits for peripheral nerve regeneration. Biomaterials 20: 1109-1115. doi: 10.1016/S0142-9612(99)00010-1
    [20] Yu Y, Meng D, Man L, et al. (2016) The Interactions Between Aligned Poly(L-Lactic Acid) Nanofibers and SH-SY5Y Cells In Vitro. J Nanosci Nanotechnol 16: 6407-6413. doi: 10.1166/jnn.2016.10883
    [21] da Rocha JF, da Cruz e Silva OA, Vieira SI (2015) Analysis of the amyloid precursor protein role in neuritogenesis reveals a biphasic SH-SY5Y neuronal cell differentiation model. J Neurochem 134: 288-301.
    [22] Koh HS, Yong T, Chan CK, et al. (2008) Enhancement of neurite outgrowth using nano-structured scaffolds coupled with laminin. Biomaterials 29: 3574-3582. doi: 10.1016/j.biomaterials.2008.05.014
    [23] Pina S, Vieira SI, Rego P, et al. (2010) Biological responses of brushite-forming Zn- and ZnSr- substituted beta-tricalcium phosphate bone cements. Eur Cell Mater 20: 162-177.
    [24] Ishihama Y, Oda Y, Tabata T, et al. (2005) Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein. Mol Cell Proteomics 4: 1265-1272. doi: 10.1074/mcp.M500061-MCP200
    [25] Mi H, Thomas P (2009) PANTHER pathway: an ontology-based pathway database coupled with data analysis tools. Methods Mol Biol 563: 123-140. doi: 10.1007/978-1-60761-175-2_7
    [26] Milacic M, Haw R, Rothfels K, et al. (2012) Annotating cancer variants and anti-cancer therapeutics in reactome. Cancers (Basel) 4: 1180-1211. doi: 10.3390/cancers4041180
    [27] Fabregat A, Sidiropoulos K, Garapati P, et al. (2016) The Reactome pathway Knowledgebase. Nucleic Acids Res 44: D481-D487. doi: 10.1093/nar/gkv1351
    [28] Henriques AG, Vieira SI, Crespo-López ME, et al. (2009) Intracellular sAPP retention in response to Abeta is mapped to cytoskeleton-associated structures. J Neurosci Res 87: 1449-1461. doi: 10.1002/jnr.21959
    [29] Romero-Calvo I, Ocón B, Martínez-Moya P, et al. (2010) Reversible Ponceau staining as a loading control alternative to actin in Western blots. Anal Biochem 401: 318-320. doi: 10.1016/j.ab.2010.02.036
    [30] Nistor G, Poole AJ, Draelos Z, et al. (2016) Human Stem Cell-Derived Skin Progenitors Produce Alpha 2-HS Glycoprotein (Fetuin): A Revolutionary Cosmetic Ingredient. J Drugs Dermatol 15: 583-598.
    [31] Elsas J, Sellhaus B, Herrmann M, et al. (2013) Fetuin-a in the developing brain. Dev Neurobiol 73: 354-369. doi: 10.1002/dneu.22064
    [32] Werbowetski-Ogilvie TE, Agar NY, Waldkircher de Oliveira RM, et al. (2006) Isolation of a natural inhibitor of human malignant glial cell invasion: inter alpha-trypsin inhibitor heavy chain 2. Cancer Res 66: 1464-1472. doi: 10.1158/0008-5472.CAN-05-1913
    [33] Ono S (2007) Mechanism of depolymerization and severing of actin filaments and its significance in cytoskeletal dynamics. Int Rev Cytol 258: 1-82. doi: 10.1016/S0074-7696(07)58001-0
    [34] Shekhar S, Pernier J, Carlier MF (2016) Regulators of actin filament barbed ends at a glance. J Cell Sci 129: 1085-1091. doi: 10.1242/jcs.179994
    [35] Barlat I, Maurier F, Duchesne M, et al. (1997) A role for Sam68 in cell cycle progression antagonized by a spliced variant within the KH domain. J Biol Chem 272: 3129-3132.
    [36] Tsuji T, Ficarro SB, Jiang W (2006) Essential role of phosphorylation of MCM2 by Cdc7/Dbf4 in the initiation of DNA replication in mammalian cells. Mol Biol Cell 17: 4459-4472.
    [37] De Vos WH, Houben F, Hoebe RA, et al. (2010) Increased plasticity of the nuclear envelope and hypermobility of telomeres due to the loss of A-type lamins. Biochim Biophys Acta 1800: 448-458.
    [38] Nardella M, Guglielmi L, Musa C, et al. (2015) Down-regulation of the Lamin A/C in neuroblastoma triggers the expansion of tumor initiating cells. Oncotarget 6: 32821-32840.
    [39] Jurica MS, Licklider LJ, Gygi SR, et al. (2002) Purification and characterization of native spliceosomes suitable for three-dimensional structural analysis. RNA 8: 426-439. doi: 10.1017/S1355838202021088
    [40] Weidensdorfer D, Stöhr N, Baude A, et al. (2009) Control of c-myc mRNA stability by IGF2BP1-associated cytoplasmic RNPs. RNA 15: 104-115.
    [41] Nami B, Ghasemi-Dizgah A, Vaseghi A (2016) Overexpression of molecular chaperons GRP78 and GRP94 in CD44(hi)/CD24(lo) breast cancer stem cells. Bioimpacts 6: 105-110. doi: 10.15171/bi.2016.15
    [42] Zhou H, Zhang Y, Fu Y, et al. (2011) Novel mechanism of anti-apoptotic function of 78-kDa glucose-regulated protein (GRP78): endocrine resistance factor in breast cancer, through release of B-cell lymphoma 2 (BCL-2) from BCL-2-interacting killer (BIK). J Biol Chem 286: 25687-25696.
    [43] Boulares AH, Yakovlev AG, Ivanova V, et al. (1999) Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis: Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem 274: 22932-22940.
    [44] Ahel I, Ahel D, Matsusaka T, et al. (2008) Poly(ADP-ribose)-binding zinc finger motifs in DNA repair/checkpoint proteins. Nature 451: 81-85. doi: 10.1038/nature06420
    [45] Moosmann P, Georgiev O, Le Douarin B, et al. (1996) Transcriptional repression by RING finger protein TIF1 beta that interacts with the KRAB repressor domain of KOX1. Nucleic Acids Res 24: 4859-48567. doi: 10.1093/nar/24.24.4859
    [46] Riclet R, Chendeb M, Vonesch JL, et al. (2009) Disruption of the interaction between transcriptional intermediary factor 1{beta} and heterochromatin protein 1 leads to a switch from DNA hyper- to hypomethylation and H3K9 to H3K27 trimethylation on the MEST promoter correlating with gene reactivation. Mol Biol Cell 20: 296-305. doi: 10.1091/mbc.E08-05-0510
    [47] Shim KS, Lubec G. Drebrin, a dendritic spine protein, is manifold decreased in brains of patients with Alzheimer's disease and Down syndrome. Neurosci Lett 324: 209-212.
    [48] Ponuwei GA (2016) A glimpse of the ERM proteins. J Biomed Sci 23: 35. doi: 10.1186/s12929-016-0246-3
    [49] Liu CX, Xu X, Chen XL, et al. (2015) Glutamate promotes neural stem cell proliferation by increasing the expression of vascular endothelial growth factor of astrocytes in vitro. Cell Mol Biol 61: 75-84.
    [50] Semrad TJ, Mack PC (2012) Fibroblast growth factor signaling in non-small-cell lung cancer. Clin Lung Cancer 13: 90-95. doi: 10.1016/j.cllc.2011.08.001
    [51] Yano S, Kondo K, Yamaguchi M, et al. (2003) Distribution and function of EGFR in human tissue and the effect of EGFR tyrosine kinase inhibition. Anticancer Res 23: 3639-3650.
    [52] Mizuguchi Y, Specht S, Isse K, et al. (2015) Breast tumor kinase/protein tyrosine kinase 6 (Brk/PTK6) activity in normal and neoplastic biliary epithelia. J Hepatol 63: 399-407. doi: 10.1016/j.jhep.2015.02.047
    [53] Lee S, Suh GY, Ryter SW, et al. (2016) Regulation and Function of the Nucleotide Binding Domain Leucine-Rich Repeat-Containing Receptor, Pyrin Domain-Containing-3 Inflammasome in Lung Disease. Am J Respir Cell Mol Biol 54: 151-160. doi: 10.1165/rcmb.2015-0231TR
    [54] Goshima Y, Yamashita N, Nakamura F, et al. (2016) Regulation of dendritic development by Semaphorin 3A through novel intracellular remote signaling. Cell Adh Migr 8: 1-14.
    [55] Mo XM, Xu CY, Kotaki M, et al. (2004) Electrospun P(LLA-CL) nanofiber: a biomimetic extracellular matrix for smooth muscle cell and endothelial cell proliferation. Biomaterials 25: 1883-1890. doi: 10.1016/j.biomaterials.2003.08.042
    [56] da Silva JS, Dotti CG (2002) Breaking the neuronal sphere: regulation of the actin cytoskeleton in neuritogenesis. Nat Rev Neurosci 3: 694-704. doi: 10.1038/nrn918
    [57] Siegel G, Agranoff B, Albers R, et al. (1999) Basic Neurochemistry: Molecular, Cellular and Medical Aspects 6 ed. Philadelphia: Lippincott-Raven.
    [58] Estefanía MM, Ganier O, Hernández P, et al. (2012) DNA replication fading as proliferating cells advance in their commitment to terminal differentiation. Sci Rep 2: 279.
    [59] Tzoneva R, Faucheux N, Groth T (2007) Wettability of substrata controls cell-substrate and cell-cell adhesions. Biochim Biophys Acta 1770: 1538-1547. doi: 10.1016/j.bbagen.2007.07.008
    [60] Alves NM, Shi J, Oramas E, et al. (2009) Bioinspired superhydrophobic poly(L-lactic acid) surfaces control bone marrow derived cells adhesion and proliferation. J Biomed Mater Res A 91: 480-488.
    [61] Titushkin I, Cho M (2009) Regulation of cell cytoskeleton and membrane mechanics by electric field: role of linker proteins. Biophys J 96: 717-728. doi: 10.1016/j.bpj.2008.09.035
    [62] Wen X, Tresco PA (2006) Effect of filament diameter and extracellular matrix molecule precoating on neurite outgrowth and Schwann cell behavior on multifilament entubulation bridging device in vitro. J Biomed Mater Res A 76: 626-637.
    [63] Weitzdoerfer R, Fountoulakis M, Lubec G (2001) Aberrant expression of dihydropyrimidinase related proteins-2,-3 and -4 in fetal Down syndrome brain. J Neural Transm Suppl 61: 95-107.
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