Preparation of NO-Releasing electrospun chitosan nanofibrous scaffolds for osteoconduction

Bone tissue engineering; chitosan; drug delivery; nanofibers; osteoconduction; sodium nitroprusside (SNP); Aqueous environment; Electrospuns; FTIR spectroscopy; Nanofibrous scaffolds; Osteoconduction; Photo-cross-linkings; SEM image; Sodium nitroprusside; Materials Science (all); Condensed Matter Physics; Mechanics of Materials; Mechanical Engineering

Abstract :

[en] This study focuses on fabricating sodium nitroprusside-releasing chitosan-based (CS/SNP) nanofibers through electrospinning. Based on SEM images and FTIR spectroscopy, the application of one-step photocrosslinking significantly improved the stability of the nanofibers in aqueous environments. SEM images showed that the porous nanofibrous structure was maintained for up to 24 h. Incorporating SNP into the nanofibrous scaffolds demonstrated an enhanced biocompatibility with osteogenic cells. The cell viability increased with rising SNP content. Enhanced cell attachment, spreading, and proliferation were also observed through fluorescence microscope images of CS/SNP nanofibers, followed by the positive regulation of Smad and Runx2 pathway. The crosstalk between CS and NO reduced the possible toxicity and enhanced the osteoinductivity of NO. Furthermore, the nanofibers contributed to escalated osteogenic differentiation and mineralization, as evidenced by heightened expression of osteogenic markers such as alkaline phosphatase (ALP) expression and calcium deposition. Overall, the photo-crosslinked electrospun CS/SNP nanofibers demonstrate substantial potential as scaffolds in the field of bone tissue engineering.

Disciplines :

Chemistry

Author, co-author :

Chiu, Chun-Yuan; Department of Orthodontics and Pediatrics Dentistry, Tri-Service General Hospital, Taipei, Taiwan ; School of Dentistry, National Defense Medical Center, Taipei, Taiwan

Lumapat, Paul Noel ; Université de Liège - ULiège > GIGA

Hsu, Yu-Tung; Department of Pharmacy, National Taiwan University, Taipei, Taiwan

Van Hong Thien, Doan; Department of Chemical Engineering, Can Tho University, Can Tho, Viet Nam

Li, Chung-Hsing; Department of Orthodontics and Pediatrics Dentistry, Tri-Service General Hospital, Taipei, Taiwan ; School of Dentistry, National Defense Medical Center, Taipei, Taiwan

Ho, Ming-Hua; Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan ; R&D Center for Membrane Technology, National Taiwan University of Science and Technology, Taipei, Taiwan

Language :

English

Title :

Preparation of NO-Releasing electrospun chitosan nanofibrous scaffolds for osteoconduction

Publication date :

2024

Journal title :

Materials Technology: Advanced Performance Materials

ISSN :

1066-7857

eISSN :

1753-5557

Publisher :

Taylor and Francis Ltd.

Volume :

Issue :

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 03 July 2025

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Bibliography

Johnell O, Kanis JA. An estimate of the worldwide prevalence and disability associated with osteoporotic fractures. Osteoporosis Int. 2006 Dec;17(12):1726–33. doi: 10.1007/s00198-006-0172-4
Shaw CK, Li YM, Wang LY, et al. Prediction of bone fracture by bone mineral density in Taiwanese. J Formos Med Assoc. 2001; 100 (12): 805–15. PMID: 11802519.
Porter JR, Ruckh TT, Popat KC. Bone tissue engineering: a review in bone biomimetics and drug delivery strategies. Biotechnol Prog. 2009; 25 (6): 1539–1560. doi: 10.1002/btpr.246
Lienemann PS, Lutolf MP, Ehrbar M. Biomimetic hydrogels for controlled biomolecule delivery to augment bone regeneration. Adv Drug Deliv Rev. 2012; 64 (12): 1078–1089. doi: 10.1016/j.addr.2012.03.010
Yilgor P, Tuzlakoglu K, Reis RL, et al. Incorporation of a sequential BMP-2/BMP-7 delivery system into chitosan-based scaffolds for bone tissue engineering. Biomaterials. 2009; 30 (21): 3551–3559. doi: 10.1016/j.biomaterials.2009.03.024
Srouji S, Ben-David D, Lotan R, et al. Slow-release human recombinant bone morphogenetic protein-2 embedded within electrospun scaffolds for regeneration of bone defect: in vitro and in vivo evaluation. Tissue Eng Part A. 2010; 17 (3–4): 269–277. doi: 10.1089/ten.tea.2010.0250
Kang J, Tada S, Kitajima T, et al. Immobilization of bone morphogenetic protein on DOPA- or dopamine-treated titanium surfaces to enhance osseointegration. Biomed Res Int. 2013; 2013: 265980. doi: 10.1155/2013/265980
Yun Y-P, Kim SE, Kang EY, et al. The effect of bone morphogenic protein-2 (BMP-2)-immobilizing heparinized-chitosan scaffolds for enhanced osteoblast activity. Tissue Eng Regen Med. 2013; 10 (3): 122–130. doi: 10.1007/s13770-013-0386-4
Fan J, Park H, Lee MK, et al. Adipose-derived stem cells and BMP-2 delivery in chitosan-based 3D constructs to enhance bone regeneration in a rat mandibular defect model. Tissue Eng Part A. 2014; 20 (15–16): 2169–2179. doi: 10.1089/ten.tea.2013.0523
Tejeda-Montes E, Smith KH, Rebollo E, et al. Bioactive membranes for bone regeneration applications: effect of physical and biomolecular signals on mesenchymal stem cell behavior. Acta Biomaterialia. 2014; 10 (1): 134–141. doi: 10.1016/j.actbio.2013.09.001
Li CH, Wang JW, Ho MH, et al. Immobilization of naringin onto chitosan substrates by using ozone activation. Colloids Surf B Biointerfaces. 2014; 115: 1–7. doi: 10.1016/j.colsurfb.2013.11.006
Li L, Zhou G, Wang Y, et al. Controlled dual delivery of BMP-2 and dexamethasone by nanoparticle-embedded electrospun nanofibers for the efficient repair of critical-sized rat calvarial defect. Biomaterials. 2015; 37: 218–229. doi: 10.1016/j.biomaterials.2014.10.015
Evans DM, Ralston SH. Nitric oxide and bone. J Bone Mineral Res. 1996; 11 (3): 300–305. doi: 10.1002/jbmr.5650110303
Saura M, Tarin C, Zaragoza C. Recent insights into the implication of nitric oxide in osteoblast differentiation and proliferation during bone development. ScientificWorldjournal. 2010; 10: 624–632. doi: 10.1100/tsw.2010.58
Wimalawansa SJ. Nitric oxide: novel therapy for osteoporosis. Expert Opin Pharmacother. 2008; 9 (17): 3025–3044. doi: 10.1517/14656560802197162
Diwan AD, Wang MX, Jang D, et al. Nitric oxide modulates fracture healing. J Bone Mineral Res. 2000; 15 (2): 342–351. doi: 10.1359/jbmr.2000.15.2.342
Park J-W, Means G. Reaction of drugs with sodium nitroprusside as a source of nitrosamines. Arch Pharm Res. 1991; 14 (2): 118–123. doi: 10.1007/BF02892015
Leeuwenkamp OR, van Bennekom WP, van der Mark EJ, et al. Nitroprusside, antihypertensive drug and analytical reagent. Review of (photo)stability, pharmacology and analytical properties. Pharm Weekbl Sci. 1984; 6 (4): 129–140. doi: 10.1007/BF01954040
Taylor TH, Styles M, Lamming AJ. Sodium nitroprusside as a hypotensive agent in general anaesthesia. Br J Anaesth. 1970; 42 (10): 859–864. doi: 10.1093/bja/42.10.859
Rőszer T. Nitric oxide signaling and nitrosative stress in the musculoskeletal system. In: Laher I, editor Systems biology of free radicals and antioxidants. Berlin Heidelberg: Springer; 2014. pp. 2895–2926.
Felka T, Ulrich C, Rolauffs B, et al. Nitric oxide activates signaling by c-raf, MEK, p-JNK, p38 MAPK and p53 in human mesenchymal stromal cells inhibits their osteogenic differentiation by blocking expression of Runx2. J Stem Cell Res Ther. 2014; 4 (04): e1187. doi: 10.4172/2157-7633.1000195
Aitken D, West D, Smith F, et al. Cyanide toxicity following nitroprusside induced hypotension. Can Anaesth Soc J. 1977; 24 (6): 651–660. doi: 10.1007/BF03006709
Lu Y, Slomberg DL, Schoenfisch MH. Nitric oxide-releasing chitosan oligosaccharides as antibacterial agents. Biomaterials. 2014; 35 (5): 1716–1724. doi: 10.1016/j.biomaterials.2013.11.015
Lu Y, Slomberg DL, Shah A, et al. Nitric oxide-releasing amphiphilic poly(amidoamine) (PAMAM) dendrimers as antibacterial agents. Biomacromolecules. 2013; 14 (10): 3589–3598. doi: 10.1021/bm400961r
Nablo BJ, Prichard HL, Butler RD, et al. Inhibition of implant-associated infections via nitric oxide release. Biomaterials. 2005; 26 (34): 6984–6990. doi: 10.1016/j.biomaterials.2005.05.017
Worley BV, Slomberg DL, Schoenfisch MH. Nitric oxide-releasing quaternary ammonium-modified poly(amidoamine) dendrimers as dual action antibacterial agents. Bioconjugate Chem. 2014; 25 (5): 918–927. doi: 10.1021/bc5000719
Yeom YE, Kim MA, Kim J, et al. Anti-inflammatory effects of the extract of solanum nigrum L. on an acute ear edema mouse model. Mater Technol. 2019; 34 (14): 851–857. doi: 10.1080/10667857.2019.1638671
Thierry B, Merhi Y, Silver J, et al. Biodegradable membrane-covered stent from chitosan-based polymers. J Biomed Mater Res Part A. 2005; 75A (3): 556–566. doi: 10.1002/jbm.a.30450
Lowe A, Deng W, Smith DW, et al. Coated melt-spun acrylonitrile-based suture for delayed release of nitric oxide. Mater Lett. 2014; 125: 221–223. doi: 10.1016/j.matlet.2014.03.174
Koh A, Carpenter AW, Slomberg DL, et al. Nitric oxide-releasing silica nanoparticle-doped polyurethane electrospun fibers. ACS Appl Mater Inter. 2013; 5 (16): 7956–7964. doi: 10.1021/am402044s
Koh A, Lu Y, Schoenfisch MH. Fabrication of nitric oxide-releasing porous polyurethane membranes-coated needle-type implantable glucose biosensors. Anal Chem. 2013; 85 (21): 10488–10494. doi: 10.1021/ac402312b
Liu HA, Balkus KJ. Novel delivery system for the bioregulatory agent nitric oxide. Chem Mater. 2009; 21 (21): 5032–5041. doi: 10.1021/cm901358z
Baldik Y, Diwan AD, Appleyard RC, et al. Deletion of iNOS gene impairs mouse fracture healing. Bone. 2005; 37 (1): 32–36. doi: 10.1016/j.bone.2004.10.002
Akins R, Rabolt J. Electrospinning fundamentals and applications. In: Buddy D, RatnerAllan S, HoffmanFrederick J, SchoenJack E, editors. Biomaterials science. 3rd ed. Cambridge, MA, USA: Academic Press; 2013. p. 332–339.
Aadil KR, Nathani A, Sharma CS, et al. Fabrication of biocompatible alginate-poly(vinyl alcohol) nanofibers scaffolds for tissue engineering applications. Mater Technol. 2018; 33 (8): 507–512. doi: 10.1080/10667857.2018.1473234
Chung TW, Lu Y-F, Wang H-Y, et al. Growth of human endothelial cells on different concentrations of gly-arg-gly-asp grafted chitosan surface. Artif Organs. 2003; 27 (2): 155–161. doi: 10.1046/j.1525-1594.2003.07045.x
Sadati SMM, Shahgholian-Ghahfarrokhi N, Shahrousvand E, et al. Edible chitosan/cellulose nanofiber nanocomposite films for potential use as food packaging. Mater Technol. 2021; 37 (10): 1276–1288. doi: 10.1080/10667857.2021.1934367
Lehr CM, Bouwstra JA, Schacht EH, et al. In vitro evaluation of mucoadhesive properties of chitosan and some other natural polymers. Int J Pharmaceut. 1992; 78 (1–3): 43–48. doi: 10.1016/0378-5173(92)90353-4
He P, Davis SS, Illum L. In vitro evaluation of the mucoadhesive properties of chitosan microspheres. Int J Pharmaceut. 1998; 166 (1): 75–88. doi: 10.1016/S0378-5173(98)00027-1
Wang K, Buschle-Diller G, Misra RDK. Chitosan-based injectable hydrogels for biomedical. Mater Technol. 2015; 30 (sup5): B198–B205. doi: 10.1179/17535557B15Y.000000008
Ho MH, Liao MH, Lin YL, et al. Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation. Int J Nanomedicine. 2014 Sep 9;9: 4293–4304. doi: 10.2147/IJN.S68012
Salehi F, Hossein B, Gholamreza K, et al. Chitosan promotes ROS-mediated apoptosis and S phase cell cycle arrest in triple-negative breast cancer cells: evidence for intercalative interaction with genomic DNA. RSC Adv. 2017; 7 (68): 43141–43150. doi: 10.1039/C7RA06793C
Liu D, Shu G, Jin F, et al. ROS-responsive chitosan-SS31 prodrug for AKI therapy via rapid distribution in the kidney and long-term retention in the renal tubule. Sci Adv. 2020 Oct 9;6 (41): eabb7422. doi: 10.1126/sciadv.abb7422
Vesey CJ, Batistoni GA. The determination and stability of sodium nitroprusside in aqueous solutions (determination and stability of SNP). J Clin Pharm Therapeutics. 1977; 2 (2): 105–117. doi: 10.1111/j.1365-2710.1977.tb00080.x
Park ES, Lind A-K, Dahm-Kähler P, et al. RUNX2 transcription factor regulates gene expression in luteinizing granulosa cells of rat ovaries. Mol Endocrinol. 2010; 24 (4): 846–858. doi: 10.1210/me.2009-0392
Cho J, Heuzey M-C, Bégin A, et al. Viscoelastic properties of chitosan solutions: effect of concentration and ionic strength. J Food Eng. 2006; 74 (4): 500–515. doi: 10.1016/j.jfoodeng.2005.01.047
Son WK, Youk JH, Lee TS, et al. Electrospinning of ultrafine cellulose acetate fibers: Studies of a new solvent system and deacetylation of ultrafine cellulose acetate fibers. J Polym Sci B Polym Phys. 2004; 42 (1): 5–11. doi: 10.1002/polb.10668
Mit-Uppatham C, Nithitanakul M, Supaphol P. Ultrafine electrospun polyamide-6 fibers: effect of solution conditions on morphology and average fiber diameter. Macromole Chem Phys. 2004; 205 (17): 2327–2338. doi: 10.1002/macp.200400225
Deitzel JM, Kleinmeyer J, Harris D, et al. The effect of processing variables on the morphology of electrospun nanofibers and textiles. Polymer. 2001; 42 (1): 261–272. doi: 10.1016/S0032-3861(00)00250-0
Demir MM, Yilgor I, Yilgor E, et al. Electrospinning of polyurethane fibers. Polymer. 2002; 43 (11): 3303–3309. doi: 10.1016/S0032-3861(02)00136-2
Sangsanoh P, Supaphol P. Stability improvement of electrospun chitosan nanofibrous membranes in neutral or weak basic aqueous solutions. Biomacromolecules. 2006; 7 (10): 2710–2714. doi: 10.1021/bm060286l
Holzbecher M, Knop O, Falk M. Infrared studies of water in crystalline hydrates: sodium nitroprusside dihydrate, na2[Fe(cn)5NO]•2H2O Na2[Fe(CN)5NO]•2H2O. Can J Chem. 1971; 49 (9): 1413–1424. doi: 10.1139/v71-233
Tang S, Edman L. On-demand photochemical stabilization of doping in light-emitting electrochemical cells. Electrochimica Acta. 2011; 56 (28): 10473–10478. doi: 10.1016/j.electacta.2011.01.073
Wold KA, Damodaran VB, Suazo LA, et al. Fabrication of biodegradable polymeric nanofibers with covalently attached NO donors. ACS Appl Mater Inter. 2012; 4 (6): 3022–3030. doi: 10.1021/am300383w
Worley BV, Soto RJ, Kinsley PC, et al. Active release of nitric xxide-releasing dendrimers from electrospun polyurethane fibers. ACS Biomater Sci Eng. 2016; 2 (3): 426–437. doi: 10.1021/acsbiomaterials.6b00032
Bakolitsa C, Cohen DM, Bankston LA, et al. Structural basis for vinculin activation at sites of cell adhesion. Nature. 2004; 430 (6999): 583–586. doi: 10.1038/nature02610
Keselowsky BG, Collard DM, García AJ, Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. Proc Natl Acad Sci; USA. 2005. Apr 26;102(17):5953–7. doi: 10.1073/pnas.0407356102
Haÿ E, Lemonnier L, Modrowski D, et al. N- and E-cadherin mediate early human calvaria osteoblast differentiation promoted by bone morphogenetic protein-2. J Cell Physiol. 2000; Apr 183 (1): 117–128. doi: 10.1002
Civitelli R. Cell–cell communication in the osteoblast/osteocyte lineage. Arch Biochem Biophys. 2008; 473 (2): 188–192. doi: 10.1016/j.abb.2008.04.005
Stains JP, Civitelli R. Cell-to-cell interactions in bone. Biochem Biophys Res Commun. 2005; 328 (3): 721–727. doi: 10.1016/j.bbrc.2004.11.078
Kim SM, Yuen T, Iqbal J, et al. The NO–cGMP–PKG pathway in skeletal remodeling. Ann N Y Acad Sci. 2021; 1487 (1): 21–30. doi: 10.1111/nyas.14486
Lai CF, Chaudhary L, Fausto A, et al. Erk is essential for growth, differentiation, integrin expression, and cell function in human osteoblastic cells. J Biol Chem. 2001; 276 (17): 14443–14450. doi: 10.1074/jbc.M010021200
Rangaswami H, Marathe N, Zhuang S, et al. Type II cGMP-dependent protein kinase mediates osteoblast mechanotransduction. J Biol Chem. 2009; 284 (22): 14796–14808. doi: 10.1074/jbc.M806486200
Huang RL, Yuan Y, Tu and J, et al. Opposing TNF-α/IL-1β- and BMP-2-activated MAPK signaling pathways converge on Runx2 to regulate BMP-2-induced osteoblastic differentiation. Cell Death Dis. 2014; 5 (4): e1187. doi: 10.1038/cddis.2014.101