Abstract
In this work, biocompatibility of bacterial cellulose (BC) was assessed as the scaffold for corneal stroma replacement. Biocompatibility was evaluated by examining rabbit corneal epithelial and stromal cells cultured on the BC scaffold. The growth of primary cells was assessed by optical microscope, scanning electron microscope (SEM), and transmission electron microscope (TEM). Live/dead viability/cytotoxicity assay and CCK-8 assay were used to evaluate cell survival. BC was surgically implanted in vivo into a stromal pocket. During a 3-month follow-up, the biocompatibility of BC was assessed. We found that epithelial and stromal cells grew well on BC and showed a survival rate of nearly 100%. The SEM examination for both kinds of cell showed abundant leafy protrusions, spherical projections, filopodia, cytoskeletons, and cellular interconnections. The stromal cells cultured on BC arranged regularly. TEM observation revealed normal cellular microstructure and a tight adhesion to the BC membrane. In vivo observation confirmed the optical transparency of BC during 3-month follow-up. The results demonstrated that BC had good biocompatibility for the tissue engineering of corneal stroma.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BC:
-
Bacterial cellulose
- SEM:
-
Scanning electron microscope
- TEM:
-
Transmission electron microscope
References
Algvere PV, Torstensson P, Tengroth B (1993) Light transmittance of ocular media in living rabbit eyes. Invest Ophthalmol Vis Sci 34(2):349–354
Dugan JM, Gough JE, Eichhorn SJ (2013) Bacterial cellulose scaffolds and cellulose nanowhiskers for tissue engineering. Nanomedicine 8(2):287–298. https://doi.org/10.2217/nnm.12.211
Fang B, Wan Y-Z, Tang T-T, Gao C, Dai K-R (2009) Proliferation and osteoblastic differentiation of human bone marrow stromal cells on hydroxyapatite/bacterial cellulose nanocomposite scaffolds. Tissue Eng A 15(5):1091–1098. https://doi.org/10.1089/ten.tea.2008.0110
Fink H, Hong J, Drotz K, Risberg B, Sanchez J, Sellborn A (2011) An in vitro study of blood compatibility of vascular grafts made of bacterial cellulose in comparison with conventionally-used graft materials. J Biomed Mater Res A 97A(1):52–58. https://doi.org/10.1002/jbm.a.33031
Gonçalves S, Padrão J, Rodrigues IP, Silva JP, Sencadas V, Lanceros-Mendez S, Girão H, Dourado F, Rodrigues LR (2015) Bacterial cellulose as a support for the growth of retinal pigment epithelium. Biomacromol 16(4):1341–1351. https://doi.org/10.1021/acs.biomac.5b00129
Hariya T, Tanaka Y, Yokokura S, Nakazawa T (2016) Transparent, resilient human amniotic membrane laminates for corneal transplantation. Biomaterials 101:76–85
Hong L, Wang YL, Jia SR, Huang Y, Gao C, Wan YZ (2006) Hydroxyapatite/bacterial cellulose composites synthesized via a biomimetic route. Mater Lett 60(13–14):1710–1713
Jun S-H, Lee S-H, Kim S, Park S-G, Lee C-K, Kang N-K (2017) Physical properties of TEMPO-oxidized bacterial cellulose nanofibers on the skin surface. Cellulose 24(12):5267–5274. https://doi.org/10.1007/s10570-017-1508-2
Keskin Z, Sendemir Urkmez A, Hames EE (2017) Novel keratin modified bacterial cellulose nanocomposite production and characterization for skin tissue engineering. Mater Sci Eng C 75:1144–1153. https://doi.org/10.1016/j.msec.2017.03.035
Kim DK, Sim BR, Kim JI, Khang G (2018) Functionalized silk fibroin film scaffold using β-Carotene for cornea endothelial cell regeneration. Colloid Surf B 164:340–346. https://doi.org/10.1016/j.colsurfb.2017.11.052
Klemm D, Schumann D, Udhardt U, Marsch S (2001) Bacterial synthesized cellulose-artificial blood vessels for microsurgery. Prog Polym Sci 26(9):1561–1603. https://doi.org/10.1016/S0079-6700(01)00021-1
Lai J-Y, Li Y-T, Cho C-H, Yu T-C (2012) Nanoscale modification of porous gelatin scaffolds with chondroitin sulfate for corneal stromal tissue engineering. Int J Nanomed 7:1101
Larkin D, Calder V, Lightman S (1997) Identification and characterization of cells infiltrating the graft and aqueous humour in rat corneal allograft rejection. Clin Exp Immunol 107(2):381–391
Liang H, Huang Y, He F, Ding HF, Wan YZ (2007) Enhanced calcium phosphate precipitation on the surface of mg-ion-implanted ZrO2 bioceramic. Surf Rev Lett 14(01):71–77. https://doi.org/10.1142/s0218625x07009086
Liu H, Liu D, Yao F, Wu Q (2010) Fabrication and properties of transparent polymethylmethacrylate/cellulose nanocrystals composites. Bioresour Technol 101(14):5685–5692
Liu X-Y, Chen J, Zhou Q, Wu J, Zhang X-L, Wang L, Qin X-Y (2013) In vitro tissue engineering of lamellar cornea using human amniotic epithelial cells and rabbit cornea stroma. Int J Ophthalmol 6(4):425–429. https://doi.org/10.3980/j.issn.2222-3959.2013.04.03
Liu Y, Lv H, Ren L, Xue G, Wang Y (2016) Improving the moisturizing properties of collagen film by surface grafting of chondroitin sulfate for corneal tissue engineering. J Biomater Sci Polym Ed 27(8):758–772
Luo H, Gu F, Xiong G, Hu D, Zhu Y, Wan Y (2016) A multichanneled bacterial cellulose scaffold for 3D in vitro cancer culture. Cell Chem Technol 50:49–56
Luo H, Dong J, Yao F, Yang Z, Li W, Wang J, Xu X, Hu J, Wan Y (2018) Layer-by-layer assembled bacterial cellulose/graphene oxide hydrogels with extremely enhanced mechanical properties. Nano Micro Lett 10(3):42. https://doi.org/10.1007/s40820-018-0195-3
Miyashita H, Shimmura S, Kobayashi H, Taguchi T, Asano-Kato N, Uchino Y, Kato M, Shimazaki J, Tanaka J, Tsubota K (2006) Collagen-immobilized poly (vinyl alcohol) as an artificial cornea scaffold that supports a stratified corneal epithelium. J Biomed Mater Res B 76B(1):56–63. https://doi.org/10.1002/jbm.b.30332
Niu G, Choi J-S, Wang Z, Skardal A, Giegengack M, Soker S (2014) Heparin-modified gelatin scaffolds for human corneal endothelial cell transplantation. Biomaterials 35(13):4005–4014. https://doi.org/10.1016/j.biomaterials.2014.01.033
Ono T, Ishiyama S, Hayashidera T, Mori Y, Nejima R, Miyata K, Amano S (2017) Twelve-year follow-up of penetrating keratoplasty. Jpn J Ophthalmol 61(2):131–136. https://doi.org/10.1007/s10384-016-0489-2
Pértile RAN, Moreira S, Gil da Costa RM, Correia A, Guãrdao L, Gartner F, Vilanova M, Gama M (2012) Bacterial cellulose: long-term biocompatibility studies. J Biomater Sci Polym Ed 23(10):1339–1354. https://doi.org/10.1163/092050611X581516
Picheth GF, Pirich CL, Sierakowski MR, Woehl MA, Sakakibara CN, de Souza CF, Martin AA, da Silva R, de Freitas RA (2017) Bacterial cellulose in biomedical applications: a review. Int J Biol Macromol 104:97–106. https://doi.org/10.1016/j.ijbiomac.2017.05.171
Rafat M, Li F, Fagerholm P, Lagali NS, Watsky MA, Munger R, Matsuura T, Griffith M (2008) PEG-stabilized carbodiimide crosslinked collagen–chitosan hydrogels for corneal tissue engineering. Biomaterials 29(29):3960–3972. https://doi.org/10.1016/j.biomaterials.2008.06.017
Saska S, Scarel-Caminaga RM, Teixeira LN, Franchi LP, dos Santos RA, Gaspar AMM, de Oliveira PT, Rosa AL, Takahashi CS, Messaddeq Y, Ribeiro SJL, Marchetto R (2012) Characterization and in vitro evaluation of bacterial cellulose membranes functionalized with osteogenic growth peptide for bone tissue engineering. J Mater Sci Mater M 23(9):2253–2266. https://doi.org/10.1007/s10856-012-4676-5
Sepúlveda RV, Valente FL, Reis ECC, Araújo FR, Eleotério RB, Queiroz PVS, Borges APB (2016) Bacterial cellulose and bacterial cellulose/polycaprolactone composite as tissue substitutes in rabbits’ cornea. Pesquisa Veterinária Brasileira 36:986–992
Svensson A, Nicklasson E, Harrah T, Panilaitis B, Kaplan DL, Brittberg M, Gatenholm P (2005) Bacterial cellulose as a potential scaffold for tissue engineering of cartilage. Biomaterials 26(4):419–431. https://doi.org/10.1016/j.biomaterials.2004.02.049
Tazi N, Zhang Z, Messaddeq Y, Almeida-Lopes L, Zanardi LM, Levinson D, Rouabhia M (2012) Hydroxyapatite bioactivated bacterial cellulose promotes osteoblast growth and the formation of bone nodules. AMB Express 2(1):61. https://doi.org/10.1186/2191-0855-2-61
Tonsomboon K, Oyen ML (2013) Composite electrospun gelatin fiber-alginate gel scaffolds for mechanically robust tissue engineered cornea. J Mech Behav Biomed Mater 21:185–194. https://doi.org/10.1016/j.jmbbm.2013.03.001
Ullah H, Wahid F, Santos HA, Khan T (2016) Advances in biomedical and pharmaceutical applications of functional bacterial cellulose-based nanocomposites. Carbohyd Polym 150:330–352. https://doi.org/10.1016/j.carbpol.2016.05.029
Wan YZ, Huang Y, He F, Wang YL, Zhao ZG, Ding HF (2006) Effect of Mg ion implantation on calcium phosphate formation on titanium. Surf Coat Technol 201(6):2904–2909
Wan YZ, Huang Y, He F, Li QY, Lian JJ (2007a) Tribological properties of three-dimensional braided carbon/Kevlar/epoxy hybrid composites under dry and lubricated conditions. Mater Sci Eng A 452:202–209
Wan YZ, Huang Y, Yuan CD, Raman S, Zhu Y, Jiang HJ, He F, Gao C (2007b) Biomimetic synthesis of hydroxyapatite/bacterial cellulose nanocomposites for biomedical applications. Mater Sci Eng C 27(4):855–864. https://doi.org/10.1016/j.msec.2006.10.002
Wan YZ, Gao C, Luo HL, He F, Liang H, Li XL, Wang YL (2009) Early growth of nano-sized calcium phosphate on phosphorylated bacterial cellulose nanofibers. J Nanosci Nanotechnol 9(11):6494–6500. https://doi.org/10.1166/jnn.2009.1311
Wan Y, Yang Z, Xiong G, Guo R, Liu Z, Luo H (2015) Anchoring Fe3O4 nanoparticles on three-dimensional carbon nanofibers toward flexible high-performance anodes for lithium-ion batteries. J Power Sources 294:414–419
Wang J, Gao C, Zhang Y, Wan Y (2010) Preparation and in vitro characterization of BC/PVA hydrogel composite for its potential use as artificial cornea biomaterial. Mater Sci Eng C 30(1):214–218. https://doi.org/10.1016/j.msec.2009.10.006
Xiao X, Pan S, Liu X, Zhu X, Connon CJ, Wu J, Mi S (2014) In vivo study of the biocompatibility of a novel compressed collagen hydrogel scaffold for artificial corneas. J Biomed Mater Res A 102(6):1782–1787. https://doi.org/10.1002/jbm.a.34848
Xiong G, Luo H, Zhang C, Zhu Y, Wan Y (2015) Enhanced biological behavior of bacterial cellulose scaffold by creation of macropores and surface immobilization of collagen. Macromol Res 23(8):734–740
Acknowledgments
This work is supported by the National Natural Science Foundation of China (Grant Nos. 81200663, 51572187, 51973058, and 31870963), Tianjin Clinical Key Discipline Project (Grant No. TJLCZDXKM014), and Key Research and Development Program of Jiangxi Province (No. 20192ACB80008, 20171BBG70112).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Zhang, C., Cao, J., Zhao, S. et al. Biocompatibility evaluation of bacterial cellulose as a scaffold material for tissue-engineered corneal stroma. Cellulose 27, 2775–2784 (2020). https://doi.org/10.1007/s10570-020-02979-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10570-020-02979-0