Nanoparticles are widely recognized for their ability to cross biological membranes and modulate cellular functions. In this study, we developed and characterized nanocomposite hydrogels composed of nanochitin (NC) embedded within a PEO/PVA polymer matrix. The materials were physicochemically characterized using zetasizer analysis, FTIR–ATR spectroscopy, and SEM imaging. Their cytotoxic potential was assessed in vitro using MTT assays on normal adult human dermal fibroblast (HDFa) cells, while developmental and neuromuscular toxicities were evaluated in vivo using Drosophila melanogaster as a model organism. Our findings indicate that all NC-doped PEO/PVA nanocomposites were non-toxic to HDFa cells. In fact, 1%, 2.5% and 5% NC-doped PEO/PVA samples showed 44.6%, 20.7% and 13.7% increases compared to negative control, respectively (p< 0.05). However, developmental toxicity assays revealed significant reductions in pupariation and survival rates, with the exception of the PEO/PVA/NC (1%) sample, which showed no adverse effects on the eclosion process. Neuromuscular assays, including larval crawling and adult climbing tests, demonstrated that while larval locomotion remained unaffected across all samples, climbing ability was significantly reduced in the 2.5% and 5% NC- biomaterial-doped groups.
ReveteAAparicioACisternaBA, et alAdvancements in the use of hydrogels for regenerative medicine: properties and biomedical applications. Int J Biomater2022; 2022: 1–16.
2.
Dhara (Ganguly)M. Polymer hydrogels: classification and recent advances. J Macromol Sci Part A2024; 61(5): 265–288.
3.
GaharwarAKPeppasNAKhademhosseiniA.Nanocomposite hydrogels for biomedical applications. Biotechnol Bioeng2014; 111(3): 441–453.
4.
BalasubramaniMKumarTRBabuM.A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications. Burns2001; 27, 5: 534–544.
5.
YildirimerLThanhNTKSeifalianMA.Evaluation of healing activity of PVA/chitosan hydrogels on deep second degree burn: pharmacological and toxicological tests. Trends Biotechnol2012; 30, 12: 638–648.
6.
BroughtonGJanisJEAttingerCE.Carboxyl-modified poly(vinyl alcohol) crosslinked chitosan hydrogel films for potential wound dressing. Plast Reconstr Surg2006; 117, 7: 12–34.
7.
VeithAPHendersonKSpencerA, et al, Fast skin healing chitosan/PEO hydrogels: in vitro and in vivo studies. Adv Drug Deliv Rev2019; 146: 97–125.
8.
NguyenHMLeTt NGOCNguyenAT, et alPoly(vinyl alcohol)/cellulose nanowhiskers nanocomposite hydrogels for potential wound dressings. RSC Adv2023; 13, 8: 5509–5528.
9.
WinterGD.High moisture strength of cassava starch/polyvinyl alcohol-compatible blends for the packaging and agricultural sectors. Nature1962; 193, 4812: 293–294.
10.
Skórkowska-TelichowskaKCzemplikMKulmaA, et alBiosensors based on porous cellulose nanocrystal–poly(vinyl alcohol) scaffolds. J Am Acad Dermatol2013; 68: 4.
11.
KamounEAKenawyERSChenX.Reproductive and developmental toxicity of carbon-based nanomaterials: a literature review. J Adv Res2017; 8, 3: 217–233.
12.
BakerMIWalshSPSchwartzZ, et alChitin and Chitosan nanocomposites for tissue engineering. J Biomed Mater Res Part B: Appl Biomater2012; 100B, 5: 1451–1457.
13.
KhodjaANMahlousMTahtatD, et alAdaptation of Drosophila melanogaster to unfavorable growth medium affects lifespan and age-related fecundity. Burns2013; 39, 1: 98 104.
14.
ZhangDZhouWWeiB, et alCarboxyl-modified poly(vinyl alcohol)-crosslinked chitosan hydrogel films for potential wound dressing. Carbohydr Polym2015; 125: 189–199. 2015.
15.
MoanessMKamelAMSalamaA, et alFast skin healing chitosan/PEO hydrogels: in vitro and in vivo studies. Int J Biol Macromol2024; 265: 130950.
16.
GonzalezJSLudueñaLNPonceA, et alPoly(vinyl alcohol)/cellulose nanowhiskers nanocomposite hydrogels for potential wound dressings. Mater Sci Eng C2014; 34: 54–61.
17.
GuimarãesMBotaroVRNovackKM, et alHigh moisture strength of cassava starch/polyvinyl alcohol-compatible blends for the packaging and agricultural sectors. J Polym Res2015; 22: 1–18. z.
18.
LinNDufresneA.Supramolecular hydrogels from in situ host-guest inclusion between chemically modified cellulose nanocrystals and cyclodextrin. Biomacromolecules2013; 14(3): 871–880.
19.
SchyrrBPascheSVoirinG, et alBiosensors based on porous cellulose nanocrystal–poly(vinyl alcohol) scaffolds. ACS Appl Mater Interfaces2014; 6(15): 12674–12683.
20.
SinghRShitizKSinghA.Nanochitin: an update review on advances in preparation methods and food applications. Int Wound J2017; 14, 6: 1276–1289.
21.
MaticaMRomanDOstafeV, et alChitosan-aluminum monostearate composite sponge dressing containing asiaticoside for wound healing and angiogenesis promotion in chronic wound. J Serb Chem Soc2023; 88, 3: 251–265.
22.
MahantaAKMaitiP.Chitin and chitosan nanocomposites for tissue engineering. In: Dutta PK (Ed.) Chitin and Chitosan for regenerative medicine. SpringerIndia, 2016, pp.123–149.
23.
EmaMHougaardKSKishimotoA, et alReproductive and developmental toxicity of carbon-based nanomaterials: a literature review. Nanotoxicology2016; 10(4): 391–412.
24.
PhaechamudTYodkhumKCharoenteeraboonJ, et alChitosan-aluminum monostearate composite sponge dressing containing asiaticoside for wound healing and angiogenesis promotion in chronic wound. Mater Sci Eng C2015; 50: 210–225.
25.
YakovlevaEUNaimarkEBMarkovAV.Adaptation of Drosophila melanogaster to unfavorable growth medium affects lifespan and age-related fecundity. Biochemistry (Mosc)2016; 81: 1445–1460.
26.
ChungHSztalTPasrichaS, et alCharacterization of Drosophila melanogaster cytochrome P450 genes. Proc Natl Acad Sci USA2009; 106(14): 5731–5736.
27.
EberlETVD. Song production in auditory mutants of Drosophila: the role of sensory feedback. J Comp Physiol A2001; c. 187(5): 341–348.
28.
ArgueKJNeckameyerWS.Sexually dimorphic recruitment of dopamine neurons into the stress response circuitry. Behav Neurosci2013; 127(5): 734–743.
29.
LiuQFLeeJHKimYM, et alIn vivo screening of traditional medicinal plants for neuroprotective activity against aβ42 cytotoxicity by using Drosophila models of Alzheimer’s disease. Biol Pharm Bull2015; 38(12): 1891–1901.
30.
GautamAGautamCMishraM, et alEnhanced mechanical properties of hBN-ZrO(2) composites and their biological activities on Drosophila melanogaster: synthesis and characterization. RSC Adv2019; 9(70): 40977–40996.
31.
MishraPKEkielskiAMukherjeeS, et alWood-based cellulose nanofibrils: haemocompatibility and impact on the development and behaviour of Drosophila melanogaster. Biomolecules2019; 9(8): 363–2019.
32.
NgasotterSSampathLXavierKAM. Nanochitin: an update review on advances in preparation methods and food applications. Carbohydr Polym2022; 291: 119627.
33.
GargSPatelPGuptaGD, et alPharmaceutical applications and advances with zetasizer: an essential analytical tool for size and zeta potential analysis. Micro Nanosystems2024; 16(3): 139–154.
34.
JiangWJTsaiMLLiuT.Chitin nanofiber as a promising candidate for improved salty taste. LWT2017; 75: 65–71.
35.
PuspawatiNMSimpenIN.Optimasi deasetilasi khitin dari kulit udang dan cangkang kepiting limbah restoran seafood menjadi khitosan melalui variasi konsentrasi NaOH. J Kim2010; 4(1): 79–90.
36.
CárdenasGCabreraGTaboadaE, et alChitin characterization by SEM, FTIR, XRD, and 13C cross polarization/mass angle spinning NMR. J Appl Polym Sci2004; 93(4): 1876–1885.
37.
KarakN.Silver nanomaterials and their polymer nanocomposites. In: Karak N (Ed.) Nanomaterials and polymer nanocomposites. Elsevier, 2019, pp.47–89. DOI: 10.1016/B978-0-12-8146156.00002-3
38.
KayaMBaranT.Description of a new surface morphology for chitin extracted from wings of cockroach (Periplaneta americana). Int J Biol Macromol2015; 75: 7–12.
39.
FreebergMATKallenbachJGAwadHA.Assessment of cellular responses of tissue constructs in vitro in regenerative engineering. Encycl Biomed Eng2019; 1: 414–426.
40.
JungOSmeetsRHartjenP, et alImproved in vitro test procedure for full assessment of the cytocompatibility of degradable magnesium based on ISO 10993-5/-12. Int J Mol Sci2019; 20(2): 255.
41.
HeMWangZCaoY, et alConstruction of chitin/PVA composite hydrogels with jellyfish Gel-Like structure and their biocompatibility. Biomacromolecules2014; 15(9): 3358–3365.
42.
IslamAYasinTGullN, et alFabrication and performance characteristics of tough hydrogel scaffolds based on biocompatible polymers. Int J Biol Macromol2016; 92: 1–10.
43.
RandMDMontgomerySLPrinceL, et alDevelopmental toxicity assays using the Drosophila model. Curr Protoc Toxicol2014; 59(1): 1.12.1–1.12.20.
44.
VecchioGGaleoneABrunettiV, et alIn vivo nanotoxicity assessment: the role of size, surface coating, nanostructuration, and dose-metrics, ech. In: Proc. 2011 NSTI Nanotechnol. Conf. Expo, NSTI-Nanotech2011, 3, pp.509–512.
45.
Silver KeySCReavesDTurnerF, et alImpacts of silver nanoparticle ingestion on pigmentation and developmental progression in Drosophila. Atlas J Biol2011; 1(3): 52–61.
46.
WangZZhangLWangX.Molecular toxicity and defense mechanisms induced by silver nanoparticles in Drosophila melanogaster. J Environ Sci2023; 125: 616–629.
47.
BarikBKMishraM.Nanoparticles as a potential teratogen: a lesson learnt from fruit fly. Nanotoxicology2019; 13(2): 258–284.
48.
AgrawalNShahP.Toxicology of nanoparticles: insights from drosophila. SpringerSingapore, 2020.
49.
PriyadarsiniSSahooSKSahuS, et alOral administration of graphene oxide nano-sheets induces oxidative stress, genotoxicity, and behavioral teratogenicity in Drosophila melanogaster. Environ Sci Pollut Res2019; 26(19): 19560–19574.
50.
SinghARajAPadmanabhanA, et alCombating silver nanoparticle-mediated toxicity in Drosophila melanogaster with curcumin. J Appl Toxicol2021; 41(8): 1188–1199.
51.
AhamedMAlSalhiMSSiddiquiMKJ. Silver nanoparticle applications and human health. Clin Chim Acta2010; 411(23-24): 1841–1848.
52.
AlarabyMAnnangiBHernándezA, et alA comprehensive study of the harmful effects of ZnO nanoparticles using Drosophila melanogaster as an in vivo model. J Hazard Mater2015; 296: 166–174.
53.
SabatDPatnaikAEkkaB, et alInvestigation of titania nanoparticles on behaviour and mechanosensory organ of Drosophila melanogaster. Physiol Behav2016; 167: 76–85.
54.
BejsovecA.Wingless/Wnt signaling in Drosophila: the pattern and the pathway. Mol Reprod Dev2013; 80(11): 882–894.
55.
EkkaBDharGSahuS, et alRemoval of Cr(VI) by silica-titania core-shell nanocomposites: in vivo toxicity assessment of the adsorbent by Drosophila melanogaster. Ceram Int2021; 47: 19079–19089.
56.
BagJMukherjeeSGhoshSK, et alFe3O4 coated guargum nanoparticles as non-genotoxic materials for biological application. Int J Biol Macromol2020; 165: 333–345.
57.
SundararajanVDanPKumarA, et alDrosophila melanogaster as an in vivo model to study the potential toxicity of cerium oxide nanoparticles. Appl Surf Sci2019; 490: 70–80.
58.
MatthewsSXuEGRoubeau DumontE, et alPolystyrene micro- and nanoplastics affect locomotion and daily activity of Drosophila melanogaster. Environ Sci Nano2021; 8(1): 110–121.
59.
ChauhanSNaikSKumarR, et alin vivo toxicological analysis of the ZnFe2O4@poly(tbge-alt-PA) nanocomposite: a study on fruit fly. ACS Omega2024; 9(6): 6549–6555.
60.
LiuXVinsonDAbtD, et alDifferential toxicity of carbon nanomaterials in Drosophila: larval dietary uptake is benign, but adult exposure causes locomotor impairment and mortality. Environ Sci Technol2009; 43: 6357–6363.
61.
ValentinoAYazdanpanahSConteR, et alSmart nanocomposite hydrogels as next-generation therapeutic and diagnostic solutions. Gels2024; 10(11): 689.
62.
CaoXXieWFengM, et alNanoplastic exposure mediates neurodevelopmental toxicity by activating the oxidative stress response in zebrafish (Danio rerio). ACS Omega2024; 9(14): 16508–16518.
63.
CocciniTPignattiPSpinilloA, et alDevelopmental neurotoxicity screening for nanoparticles using neuron-like cells of human umbilical cord mesenchymal stem cells: example with magnetite nanoparticles. Nanomater2020; 10(8): 1607.
64.
ChenSChenYGaoY, et alToxic effects and mechanisms of nanoplastics on embryonic brain development using brain organoids model. Sci Total Environ2023; 904: 166913.
65.
HuangWMoJLiJ, et alExploring developmental toxicity of microplastics and nanoplastics (MNPS): insights from investigations using zebrafish embryos. Sci Total Environ2024; 933: 173012.
