Impact of crystalline domains on long-term stability and mechanical performance of anisotropic silk fibroin sponges.
crystalline biopolymers
hysteresis
silk fibroin
tensile strength
viscoelasticity
Journal
Journal of biomedical materials research. Part A
ISSN: 1552-4965
Titre abrégé: J Biomed Mater Res A
Pays: United States
ID NLM: 101234237
Informations de publication
Date de publication:
12 Mar 2024
12 Mar 2024
Historique:
revised:
20
02
2024
received:
01
12
2023
accepted:
26
02
2024
medline:
12
3
2024
pubmed:
12
3
2024
entrez:
12
3
2024
Statut:
aheadofprint
Résumé
Sponge-like materials made from regenerated silk fibroin biopolymers are a tunable and advantageous platform for in vitro engineered tissue culture and in vivo tissue regeneration. Anisotropic, three-dimensional (3D) silk fibroin sponge-like scaffolds can mimic the architecture of contractile muscle. Herein, we use silk fibroin solution isolated from the cocoons of Bombyx mori silkworms to form aligned sponges via directional ice templating in a custom mold with a slurry of dry ice and ethanol. Hydrated tensile mechanical properties of these aligned sponges were evaluated as a function of silk polymer concentration (3% or 5%), freezing time (50% or 100% ethanol), and post-lyophilization method for inducing crystallinity (autoclaving, water annealing). Hydrated static tensile tests were used to determine Young's modulus and ultimate tensile strength across sponge formulations at two strain rates to evaluate rate dependence in the calculated parameters. Results aligned with previous reports in the literature for isotropic silk fibroin sponge-like scaffolds, where the method by which beta-sheets were formed and level of beta-sheet content (crystallinity) had the greatest impact on static parameters, while polymer concentration and freezing rate did not significantly impact static mechanical properties. We estimated the crystalline organization using molecular dynamics simulations to show that larger crystalline regions may be responsible for strength at low strain amplitudes and brittleness at high strain amplitudes in the autoclaved sponges. Within the parameters evaluated, extensional Young's modulus is tunable in the range of 600-2800 kPa. Dynamic tensile testing revealed the linear viscoelastic region to be between 0% and 10% strain amplitude and 0.2-2 Hz frequencies. Long-term stability was evaluated by hysteresis and fatigue tests. Fatigue tests showed minimal change in the storage and loss modulus of 5% silk fibroin sponges for more than 6000 min of continuous mechanical stimulation in the linear regime at 10% strain amplitude and 1 Hz frequency. Furthermore, we confirmed that these mechanical properties hold when decellularized extracellular matrix is added to the sponges and when the mechanical property assessments were performed in cell culture media. We also used nano-computed tomography (nano-CT) and simulations to explore pore interconnectivity and tortuosity. Overall, these results highlight the potential of anisotropic, sponge-like silk fibroin scaffolds for long-term (>6 weeks) contractile muscle culture with an in vitro bioreactor system that provides routine mechanical stimulation.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Science Foundation Graduate Research Fellowship
ID : DGE-2236414
Organisme : National Science Foundation Research Experience
ID : EEC-1852111
Informations de copyright
© 2024 Wiley Periodicals LLC.
Références
Stoppel WL, Hu D, Domian IJ, Kaplan DL, Black LD 3rd. Anisotropic silk biomaterials containing cardiac extracellular matrix for cardiac tissue engineering. Biomed Mater. 2015;10(3):034105. doi:10.1088/1748-6041/10/3/034105
Kukla DA, Stoppel WL, Kaplan DL, Khetani SR. Assessing the compatibility of primary human hepatocyte culture within porous silk sponges. RSC Adv. 2020;10(62):37662-37674. doi:10.1039/d0ra04954a
Joukhdar H, Och Z, Tran H, et al. Imparting multi-scalar architectural control into silk materials using a simple multi-functional ice-Templating fabrication platform. Adv Mater Technol. 2023;8(8):2201642. doi:10.1002/admt.202201642
Rnjak-Kovacina J, DesRochers TM, Burke KA, Kaplan DL. The effect of sterilization on silk fibroin biomaterial properties. Macromol Biosci. 2015;15(6):861-874. doi:10.1002/mabi.201500013
Rnjak-Kovacina J, Wray LS, Burke KA, et al. Lyophilized silk sponges: a versatile biomaterial platform for soft tissue engineering. ACS Biomater Sci Eng. 2015;1(4):260-270. doi:10.1021/ab500149p
Rnjak-Kovacina J, Wray LS, Golinski JM, Kaplan DL. Arrayed hollow channels in silk-based scaffolds provide functional outcomes for engineering critically-sized tissue constructs. Adv Funct Mater. 2014;24(15):2188-2196. doi:10.1002/adfm.201302901
Jameson JF, Pacheco MO, Bender EC, et al. Impact of bioactive molecule inclusion in lyophilized silk scaffolds varies between in vivo and in vitro assessments. bioRxiv. 2022. doi:10.1101/2022.05.24.493207
Jameson JF, Pacheco MO, Butler JE, Stoppel WL. Estimating kinetic rate parameters for enzymatic degradation of lyophilized silk fibroin sponges. Front bioeng biotechnol. 2021;9(537):664306. doi:10.3389/fbioe.2021.664306
Stoppel WL, Ghezzi CE, McNamara SL, Black LD 3rd, Kaplan DL. Clinical applications of naturally derived biopolymer-based scaffolds for regenerative medicine. Ann Biomed Eng. 2015;43(3):657-680. doi:10.1007/s10439-014-1206-2
Abbott RD, Kimmerling EP, Cairns DM, Kaplan DL. Silk as a biomaterial to support long-term three-dimensional tissue cultures. ACS Appl Mater Interfaces. 2016;8(34):21861-21868. doi:10.1021/acsami.5b12114
Abbott RD, Wang RY, Reagan MR, et al. The use of silk as a scaffold for mature, sustainable unilocular adipose 3D tissue engineered systems. Adv Healthc Mater. 2016;5(13):1667-1677. doi:10.1002/adhm.201600211
Alave Reyes-Furrer A, De Andrade S, Bachmann D, et al. Matrigel 3D bioprinting of contractile human skeletal muscle models recapitulating exercise and pharmacological responses. Commun Biol. 2021;4(1):1183. doi:10.1038/s42003-021-02691-0
Volpi M, Paradiso A, Costantini M, Swieszkowski W. Hydrogel-based fiber biofabrication techniques for skeletal muscle tissue engineering. ACS Biomater Sci Eng. 2022;8(2):379-405. doi:10.1021/acsbiomaterials.1c01145
Barton ER, Pacak CA, Stoppel WL, Kang PB. The ties that bind: functional clusters in limb-girdle muscular dystrophy. Skelet Muscle. 2020;10(1):22. doi:10.1186/s13395-020-00240-7
Stoppel WL, Kaplan DL, Black LD 3rd. Electrical and mechanical stimulation of cardiac cells and tissue constructs. Adv Drug Deliv Rev. 2016;96:135-155. doi:10.1016/j.addr.2015.07.009
Lin X, Fan L, Wang L, Filppula AM, Yu Y, Zhang H. Fabricating biomimetic materials with ice-templating for biomedical applications. Smart Med. 2023;2(3):e20230017. doi:10.1002/SMMD.20230017
Diaz F, Forsyth N, Boccaccini AR. Aligned ice templated biomaterial strategies for the musculoskeletal system. Adv Healthc Mater. 2023;12(21):e2203205. doi:10.1002/adhm.202203205
Joukhdar H, Seifert A, Jüngst T, Groll J, Lord MS, Rnjak-Kovacina J. Ice templating soft matter: fundamental principles and fabrication approaches to tailor pore structure and morphology and their biomedical applications. Adv Mater. 2021;33(34):2100091. doi:10.1002/adma.202100091
Byette F, Bouchard F, Pellerin C, Paquin J, Marcotte I, Mateescu MA. Cell-culture compatible silk fibroin scaffolds concomitantly patterned by freezing conditions and salt concentration. Polym Bull. 2011;67(1):159-175. doi:10.1007/s00289-010-0438-z
Asuncion MCT, Goh JC-H, Toh S-L. Anisotropic silk fibroin/gelatin scaffolds from unidirectional freezing. Mater Sci Eng C. 2016;67:646-656. doi:10.1016/j.msec.2016.05.087
Mao M, He J, Liu Y, Li X, Li D. Ice-template-induced silk fibroin-chitosan scaffolds with predefined microfluidic channels and fully porous structures. Acta Biomater. 2012;8(6):2175-2184. doi:10.1016/j.actbio.2011.12.025
Mandal BB, Gil ES, Panilaitis B, Kaplan DL. Laminar silk scaffolds for aligned tissue fabrication. Macromol Biosci. 2012;13(1):48-58. doi:10.1002/mabi.201200230
Hu X, Shmelev K, Sun L, et al. Regulation of silk material structure by temperature-controlled water vapor annealing. Biomacromolecules. 2011;12(5):1686-1696. doi:10.1021/bm200062a
Hu X, Kaplan D, Cebe P. Dynamic protein-water relationships during β-sheet formation. Macromolecules. 2008;41(11):3939-3948. doi:10.1021/ma071551d
Zhou CZ, Confalonieri F, Jacquet M, Perasso R, Li ZG, Janin J. Silk fibroin: structural implications of a remarkable amino acid sequence. Proteins. 2001;44(2):119-122. doi:10.1002/prot.1078
Patel M, Dubey DK, Singh SP. Phenomenological models of Bombyx mori silk fibroin and their mechanical behavior using molecular dynamics simulations. Mater Sci Eng C Mater Biol Appl. 2020;108:110414.
Zhang L, Zhang W, Hu Y, et al. Systematic review of silk scaffolds in musculoskeletal tissue engineering applications in the recent decade. ACS Biomater Sci Eng. 2021;7(3):817-840. doi:10.1021/acsbiomaterials.0c01716
Rockwood DN, Preda RC, Yucel T, Wang X, Lovett ML, Kaplan DL. Materials fabrication from Bombyx mori silk fibroin. Nat Protoc. 2011;6(10):1612-1631. doi:10.1038/nprot.2011.379
Hu X, Kaplan D, Cebe P. Determining beta-sheet crystallinity in fibrous proteins by thermal analysis and infrared spectroscopy. Macromolecules. 2006;39(18):6161-6170. doi:10.1021/ma0610109
Fossey SA, Nemethy G, Gibson KD, Scheraga HA. Conformational energy studies of beta-sheets of model silk fibroin peptides. I. Sheets of poly(ala-Gly) chains. Biopolymers. 1991;31(13):1529-1541. doi:10.1002/bip.360311309
Humphrey W, Dalke A, Schulten K. VMD: visual molecular dynamics. J Mol Graph. 1996;14(1):33-38. doi:10.1016/0263-7855(96)00018-5
Abraham MJ, Murtola T, Schulz R, et al. GROMACS: high performance molecular simulations through multi-level parallelism from laptops to supercomputers. SoftwareX. 2015;1-2:19-25. doi:10.1016/j.softx.2015.06.001
Thompson AP, Aktulga HM, Berger R, et al. LAMMPS-a flexible simulation tool for particle-based materials modeling at the atomic, meso, and continuum scales. Comput Phys Commun. 2022;271:108171. doi:10.1016/j.cpc.2021.108171
Huang J, Rauscher S, Nawrocki G, et al. CHARMM36m: an improved force field for folded and intrinsically disordered proteins. Nat Methods. 2017;14(1):71-73. doi:10.1038/nmeth.4067
Shirts MR, Klein C, Swails JM, et al. Lessons learned from comparing molecular dynamics engines on the SAMPL5 dataset. J Comput Aided Mol des. 2017;31(1):147-161. doi:10.1007/s10822-016-9977-1
Ryckaert J-P, Ciccotti G, Berendsen HJC. Numerical integration of the cartesian equations of motion of a system with constraints: molecular dynamics of n-alkanes. J Comput Phys. 1977;23(3):327-341. doi:10.1016/0021-9991(77)90098-5
Pacheco MO, Lutz HM, Armada J, et al. Silk fibroin particles as carriers in the development of hemoglobin-based oxygen carriers. Adv. NanoBiomed Res. 2023;3:2300019. doi:10.1002/anbr.202300019
Callister J, William D, Rethwisch DG. Fundamentals of Materials Science and Engineering: An Integrated Approach. 5th ed. John Wiley and Sons, Inc; 2015:216-278.
Cheng L, Shao JX, Wang F, Li Z, Dai FY. Strain rate dependent mechanical behavior of B. mori silk, A. assama silk, A. pernyi silk and A. ventricosus spider silk. Materials & Design. 2020;195:108988. doi:10.1016/j.matdes.2020.108988
Morrow DA, Haut Donahue TL, Odegard GM, Kaufman KR. Transversely isotropic tensile material properties of skeletal muscle tissue. J Mech Behav Biomed Mater. 2010;3(1):124-129. doi:10.1016/j.jmbbm.2009.03.004
Kuthe CD, Uddanwadiker RV. Investigation of effect of fiber orientation on mechanical behavior of skeletal muscle. J Appl Biomater Funct Mater. 2016;14(2):154-162. doi:10.5301/jabfm.5000275
Brown J, Lu CL, Coburn J, Kaplan DL. Impact of silk biomaterial structure on proteolysis. Acta Biomater. 2015;11:212-221. doi:10.1016/j.actbio.2014.09.013
Asakura T, Okushita K, Williamson MP. Analysis of the structure of silk fibroin by NMR. Macromolecules. 2015;48(8):2345-2357. doi:10.1021/acs.macromol.5b00160
Huang WW, Krishnaji S, Tokareva OR, Kaplan D, Cebe P. Influence of water on protein transitions: morphology and secondary structure. Macromolecules. 2014;47(22):8107-8114. doi:10.1021/ma5016227
Inoue S, Tanaka K, Arisaka F, Kimura S, Ohtomo K, Mizuno S. Silk fibroin of Bombyx mori is secreted, assembling a high molecular mass elementary unit consisting of H-chain, L-chain, and P25, with a 6:6:1 molar ratio. J Biol Chem. 2000;275(51):40517-40528. doi:10.1074/jbc.M006897200
Hossain D, Tschopp MA, Ward DK, Bouvard JL, Wang P, Horstemeyer MF. Molecular dynamics simulations of deformation mechanisms of amorphous polyethylene. Polymer. 2010;51(25):6071-6083. doi:10.1016/j.polymer.2010.10.009
Mezger TG. Oscillatory Tests. Applied Rheology: With Joe Flow on Rheology Road. 5th ed. Graz, Austria: Anton Paar GmbH. 2018. 89-100.
Jalal S, Dastidar S, Tedesco FS. Advanced models of human skeletal muscle differentiation, development and disease: three-dimensional cultures, organoids and beyond. Curr Opin Cell Biol. 2021;73:92-104. doi:10.1016/j.ceb.2021.06.004
Jiang Y, Torun T, Maffioletti SM, Serio A, Tedesco FS. Bioengineering human skeletal muscle models: recent advances, current challenges and future perspectives. Exp Cell Res. 2022;416(2):113133. doi:10.1016/j.yexcr.2022.113133
Ostrovidov S, Ramalingam M, Bae H, et al. Latest developments in engineered skeletal muscle tissues for drug discovery and development. Expert Opin Drug Discovery. 2023;18(1):47-63. doi:10.1080/17460441.2023.2160438
Mihaly E, Altamirano DE, Tuffaha S, Grayson W. Engineering skeletal muscle: building complexity to achieve functionality. Semin Cell Dev Biol. 2021;119:61-69. doi:10.1016/j.semcdb.2021.04.016
Vesga-Castro C, Aldazabal J, Vallejo-Illarramendi A, Paredes J. Contractile force assessment methods for in vitro skeletal muscle tissues. Elife. 2022;11:e77204. doi:10.7554/eLife.77204
Somers SM, Spector AA, DiGirolamo DJ, Grayson WL. Biophysical stimulation for engineering functional skeletal muscle. Tissue Eng Part B Rev. 2017;23(4):362-372. doi:10.1089/ten.TEB.2016.0444
Somers SM, Zhang NY, Morrissette-McAlmon JBF, Tran K, Mao HQ, Grayson WL. Myoblast maturity on aligned microfiber bundles at the onset of strain application impacts myogenic outcomes. Acta Biomater. 2019;94:232-242. doi:10.1016/j.actbio.2019.06.024
Somers SM, Grayson WL. Protocol for the use of a novel bioreactor system for hydrated mechanical testing, strained sterile culture, and force of contraction measurement of tissue engineered muscle constructs. Front Cell Dev Biol. 2021;9:661036. doi:10.3389/fcell.2021.661036
Qazi TH, Wu J, Muir VG, et al. Anisotropic rod-shaped particles influence injectable granular hydrogel properties and cell invasion. Adv Mater. 2022;34(12):e2109194. doi:10.1002/adma.202109194
McCuller C, Jessu R, Callahan AL. Physiology, Skeletal Muscle. StatPearls. Treasure Island (FL): StatPearls Publishing Copyright © 2023, StatPearls Publishing LLC. 2023.
Carnes ME, Pins GD. Skeletal muscle tissue engineering: biomaterials-based strategies for the treatment of volumetric muscle loss. Bioengineering (Basel). 2020;7(3):85. doi:10.3390/bioengineering7030085